Objective To investigate the feasibility of primary culture of nontumorigenic human mammary epithelial cells,and the effect of proliferation due to hormone.Methods Normal human mammary tissue was taken from mammary segment ectomy,and the tissue was digested by collagenase type Ⅰ.Epithelial depuration was carried out by integration of burning and enzyme digestion.The cells we acquired were identified by observation of cell morphology by light and Transmission Electron Microscopy(TEM).Results It is feasible to culture nontumorigenic human mammary epithelial cells.?-estradiol and progesterone at 1?10-5 g/L can promote the proliferation of the normal human mammary epithelial cells.

Objective To establish a cisplatin-resistant 4T1 mouse model of triple negative breast cancer .Meth-ods A drug resistant mice model was established with cisplatin ( DDP ) induction and in-vivo/in-vitro tumorigenic ap-proach.Its resistance characteristics were identified by MTT assay .Changes of drug resistance gene ( MDR1, BCRP, MMP7, GST-π) and protein ( P-gp, BCRP, MMP7) expression, and phosphorate-Akt and total-Akt protein expression were evaluated by real-time PCR, immunohistochemistry and western blot method , respectively.Small animal live imaging technology was applied to detect tumor growth .Results Resistance fold (RF) of cisplatin-resistant 4T1 mouse model was 12.84.The expression of MDR1, BCRP, MMP 7, GST-πmRNA and P-gp, BCRP, MMP 7 proteins in the resistant mice were higher than that in the non-resistant mice .The result of western blot showed that a statistically higher expression of p-Akt in resistant mice than that in non-resistant mice at protein levels (P<0.01).No significant difference of tumor growth rate was observed between non-resistant and resistant mice ( P>0.05 ) .Given same dose of DDP , resistant mice showed lower sensitivity than non-resistant mice significantly (P<0.01).Conclusions We have successfully established a cis-platin-resistant triple negative breast cancer model in mice , which provides a new platform for further study on chemoresis-tant reversal strategy and individualized clinical treatment of this disease .

Treatment options are limited for triple negative breast cancer (TNBC)since endocrinother-apy and targeted therapy that aims directly at human epidermal growth factor receptor-2 (HER-2)are ineffec-tive.As such,in addition to surgical treatment,the mainstay of treatment of TNBC is systemic cytotoxic chemo-therapy.The targeted therapy of TNBC is becoming a research hotspot because of traditional chemotherapy cura-tive effect is not good enough.A large number of clinical trials have found that patients with TNBC can get ben-efits from targeted molecular strategies including poly-adenosine diphosphate glucose pyrophospheralase-ribose polymerase-1 (PARP-1 )inhibitor and epidermal growth factor receptor (EGFR)inhibitor.

OBJECTIVE: To investigate the effects of Astragalus injection (AI) on basal-like breast cancer cell line MDA-MB-468 and murine bone marrow stromal stem cells (mMSCs). METHODS: MDA-MB-468 cells and primary cultured mMSCs were treated by different concentrations of AI, and with untreated MDA-MB-468 cells as blank control. The morphology of cells was observed by phase-contrast inverted microscope and transmission electron microscopy. Cytotoxic effects of AI on MDA-MB-468 cells and mMSCs were evaluated by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis of MDA-MB-468 cells induced by AI were measured by flow cytometry. Lactate dehydrogenase (LDH) activity in supernatants was measured by enzymatic colorimetric method. The expressions of epidermal growth factor receptor (EGFR) and p53 protein in MDA-MB-468 cells were evaluated by streptavidin-biotin-peroxidase complex method. RESULTS: A time-dependent cytotoxic effect of 1 g/ml AI was observed in MDA-MB-468 cells. 1 g/ml AI also had cytotoxic effect on mMSCs, but its effect was not better than cisplatin. 0.1 g/ml AI could promote the proliferation of mMSCs. Different concentrations of AI could all induce the apoptosis of MDA-MB-468 cells. There was no significant difference in LDH activity in the supernatants between blank control group and AI-treated and cisplatin-treated groups. AI could down-regulate the expressions of EGFR and p53 protein. CONCLUSION: The effects of AI on MDA-MB-468 cells and mMSCs are related to the concentration of AI, and its mechanism of inhibiting the proliferation of MDA-MB-468 cells may be due to down-regulation of the expressions of EGFR and p53 protein.

Objective: To observe the effects of Astragalus injection, astragaloside IV and formononetin on proliferation and Akt phosphorylation of basal-like human breast carcinoma cell lines MDA-MB-468 and MDA-MB-231. Methods: The effects of different concentrations of Astragalus injection, astragaloside IV and formononetin on proliferation of breast cancer cell lines were assayed by methyl thiazolyl tetrazolium (MTT) assay, and their effects on phospho-Akt were assayed by in-cell Western blot method. Results: The results of the MTT assay showed that the best concentrations of Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin were 1 g/mL, 80 mug/mL, 40 mug/mL and 10 mug/mL plus 40 mug/mL respectively. After 1- or 2-day culture, Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin decreased the expressions of p-Akt (Thr 308) and p-Akt (Ser 473) in MDA-MB-468 cells. Formononetin and astragaloside IV plus formononetin down-regulated the expression of p-Akt (Thr 308) protein in MDA-MB-231 cells after 1- and 2-day culture, but had no effects on the expression of p-Akt (Ser 473) protein in MDA-MB-231 cells. Conclusion: Astragalus injection, astragaloside IV and formononetin can inhibit proliferation of breast cancer cell lines MDA-MB-468 and MDA-MB-231, and the antiproliferation effects vary according to their concentrations. And the antiproliferation mechanisms may be related to their down-regulation effects on Akt phosphorylation.

OBJECTIVE: To evaluate the effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture. METHODS: The normal human mammary epithelial cells were dissociated by digestion with collagenase type I. The morphological identification of normal human mammary epithelial cells in primary culture was determined under inverted phase contrast microscope and transmission electron microscope. The estradiol and progesterone were added to culture medium to induce the proliferation of normal human mammary epithelial cells in primary culture, and the different doses of Chinese herbal medicine for warming kidney and tamoxifen were also added into the culture medium. The MTT assay was used to evaluate the proliferation of normal human mammary epithelial cells in primary culture under the different conditions of interventions. RESULTS: The normal human mammary epithelial cells in primary culture presented typically morphological features of normal human mammary epithelial cells in vitro. The proliferation of normal human mammary epithelial cells in primary culture was significantly improved by intervention of estradiol and progesterone both with concentration of 1x10(-5) g/L. The high-dose Chinese herbal medicine for warming kidney could obviously promote the proliferation of normal human mammary epithelial cells in primary culture, while the proliferation was obviously inhibited by low-dose Chinese herbal medicine for warming kidney. The tamoxifen exerted no effects on the proliferation of normal human mammary epithelial cells in primary culture. CONCLUSION: The estradiol and progesterone can enhance the ability of proliferation of normal human mammary epithelial cells in primary culture. The regulating effect of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture is dose-dependent.

20% ) were assigned to 2 groups according to study intervention: group A ( n =72) receiving DFPP and group B ( n =41) as controls.Group A was subdivided into 2 groups:group A1 ( n =44) was treated by DFPP alone and group A2 ( n =28) was treated by DFPP plus Dac.The incidence rates of HAR,AR,adverse reaction,patie nt/kidney survival,kidney function were observed. All the patients obtained a fo llow-up ≥12 months. Results In 72 patients of group A ,the level of PRA decreased from (60.5?17.7)% to (19.3?11.2)%,with a mean of (41.2?16.9)% ( P 0.05),with 1 kidney-year survival of 94.4% in group A and 78.0 % in group B ( P 0.05);those of AR were 36.4% and 14.3%,respectively ( P 0.05).No difference in infection episodes an d adverse events between group A and PRA-negative recipients, the same as those between group A1 and A2. Conclusions DFPP can decrease the level of PRA significantly before transplantation by selectively eliminating the sensitive antibody,especially when combined with Dac,which can make sensiti zed recipients get the chance of transplanting and further reduce the incidence of AR.The patient/kidney survival rates of 1 year are satisfactory.Being well to lerated by the sensitized patients,treatment of DFPP combined with Dac is safe a nd effective.

Objective To analyze the use of antibiotics in perioperative period of cesarean section,and to summarize the experience of scientific intervention to promote rational drug use.Method 192 cesarean section included in the study were inpatients in December 2015 to December 2016 in our hospital,were grouped according to the order of admission,divided into control group(95 cases)and observation group(97 cases),were given routine perioperative antibiotic treatment.The observation group was treated with scientific medication intervention,and the control group was not intervened.The two groups of white blood cell count,postoperative body temperature and hospitalization time were counted,the infection types and infection rate were counted,and the treatment costs of the two groups were counted.Results The white blood cell count of the observation group was significantly higher than the control group,postoperative body temperature recovery time and hospitalization time was significantly shorter than the control group,the difference was significant(P<0.05); after two groups appeared infection phenomenon,including urinary tract infection,incision infection and puerperal infection as well as intrauterine infection,and two groups in different types of infection and the infection rate by comparison there was no statistically significant difference; statistical analysis and comparison of treatment related costs of the two groups of patients,to observe antibacterial drug group,the total cost of expenses and hospitalization costs were significantly lower than that of the control group,and patients with antimicrobial drugs accounted for the total proportion of drugs were significantly lower than the control group,the difference was statistically significant(P<0.05).Conclusion Cesarean section perioperative use of antibiotics in the process,through the implementation of scientific intervention can obtain ideal effect,promote rational and normative clinical medication,reduce hospitalization expenses and antimicrobial drug costs,promote the recovery of patients after surgery,and does not increase the infection after operation.