Objective To investigate the expression of stromal cell-derived factor-1 (SDF-1) and its clinical significance in blood plasma of patients with breast tumor. Methods The level of SDF-1 protein was examined by enzyme linked immunosorbent assay (ELISA) in blood plasma of 26 patients with breast benign tumor and 52 patients with breast cancer. Results The SDF-1 protein in blood plasma was detected in both breast benign tumor patients and breast cancer ones. The level of SDF-1 protein in patients with breast cancer was higher than that in ones with breast benign tumor, and there was a statistical difference between them （P=0.000）. In patients with breast cancer, the level of SDF-1 protein in axillary lymph node (ALN) metastasis positive patients was significantly higher than that in ALN metastasis negative ones （P=0.036）. Conclusion The level of SDF-1 protein in blood plasma may be a specific tumor marker. Its level is correlated with lymph node involvement in breast cancer.
ObjectiveTo investigate the effect and mechanism of caveolion-1 on the growth and proliferation of human pancreatic carcinoma cell Panc1, in vitro. MethodsThe plasmid pCI-neo-cav-1 and its corresponding empty vector (pCI-neo) were transfected into Panc1 cell line (study group and control group, respectively). Expressions of caveolin-1, Akt, and Aktphosphate (p-Akt) were determined in transfectants by Western blot analysis. The cell growth curve was drawn and the double time was calculated in each group, and the cell cycle was analyzed by flow cytometry. The colony formation ability of tumor cells was detected by anchorageindependent growth assay. ResultsCaveolin-1 expression was up-regulated (Plt;0.01) and the growth of Panc1 cell was inhibited significantly (Plt;0.01) in the study group comparing with the control group. Caveolin-1 overexpression inhibited proliferation of Panc1 cell by arresting the cell cycle in the G0/G1 phase (Plt;0.05), the rate of S phase in the study group was lower than that of the control group (Plt;0.01). Proliferation index of the study group was also lower than that of the control group (Plt;0.01). Caveolin-1 overexpression reduced the capacity of the cells to form colonies in soft agar (Plt;0.01). p-Akt protein was reduced in the study group as compared with the control group (Plt;0.05). ConclusionCaveolin-1 can function as a cancer suppressor through inhibiting the activation of PI3K/Akt signaling pathway in Panc1 cell.
Objective To investigate the way to culture human parathyroid cells and to investigate its secretory function. Methods After digested by collagenase, parathyroid cells were isolated to get the original generation cells, then the cells were cultured and passaged, and morphological changes of original generation cells and passage cells were observed on every day. The parathyroid hormone(PTH) level secreted by the original generation cells and passage cells were measured on the 1st, 5th, 10th, 15th, and 20th day(original generation cells only) respectively. Results The cellular morphology was complete after digestion. On the 2nd day, most of the parathyroid cells had adhered and spreaded, on the 3rd day, all cells had spread. There was no very obvious changes on these cells after cultured for 4-15 days. From 16 to 20 days, some parathyroid cells went senescence. On the 1st day, all of the passage cells, which were fusiform and little bigger than those of the original generation cells, had adhered and spreaded. From 2 to 15 days, there was no very obvious changes. The concentration of PTH in original generation cells begin to decreased significantly on the 10th day (P < 0.01). The concentration of PTH in passage cells were all lower than those of original generation cells at the same corresponding time, but there were no significant difference on the PTH level on 5th day and 1st day, 10th day and 5th day, 15th day and 10th day in passage cells (P > 0.05). Conclusion Parathyroid cells which were cultured within 10 days possess well morphologic structure and have the strongest secretory function. Although the passage cells still possess secretory function, it is greatly inferior to original generation cells. At last, we consider that original generation cells cultured within 10 days can be regarded as the source of allogeneic cell transplantation.
Objective To investigate the reliability of culture method of adult human parathyroid cells. Methods Adult human parathyroid tissue was digested by collagenase, then the original generation of cells were cultured and passaged, and their morphological changes were observed and recorded every other day. Part of the passaged cells were observed through electron microscope and its supernatant parathyroid hormone (PTH) was assayed. Meanwhile, the other part of cells were tested the parathyroid markers, including PTH, calcium-sensing receptor (CaSR) and glial cells missing-2 (GCM-2) by PCR. Results Abundant cytoplasm, mitochondria, endoplasmic reticulum and Golgi apparatus from the seventh day's passaged cells were observed by the electron microscopy, as well as, some secretory granules existing in both cytoplasm and intercellular lacuna. Also, the PTH from supernate was detected, and parathyroid specific markers, such as CaSR, PTH, and GCM2 were positive. Conclusions These trials demonstrated the adult human parathyroid cells could be harvested by collagenase digestion and the cultured. Furthermore, the cells remained good shape and kept functioning, making it a potential source for allogeneic cell transplantation to the treatment of permanent hypoparathyroidism.
ObjectivesTo systematically review the efficacy and safety of ciprofloxacin for non-cystic fibrosis bronchiectasis.MethodsDatabases including PubMed, EMbase, The Cochrane Library, CBM, VIP, CNKI and WanFang Data were electronically searched from inception to August 2018 to collect randomized controlled trials (RCTs) on ciprofloxacin in the treatment of non-cystic fibrosis bronchiectasis. Two reviewers independently screened literature, extracted data, and assessed risk of bias of included studies. Then, meta-analysis was performed by using RevMan 5.3 software.ResultsA total of 9 RCTs involving 1 666 patients were included. The results of meta-analysis showed that: compared with control group, the ciprofloxacin more efficiently eradicate bacteria from sputum (RR=4.34, 95%CI 2.04 to 9.23, P=0.000 1), decrease risk of the exacerbations (RR=0.81, 95%CI 0.71 to 0.93, P=0.002) and the mean bacterial load (MD=–4.08, 95%CI –6.29 to –1.87, P=0.001). However, there were no significant differences between two groups in clinical efficiency and adverse events.ConclusionsThe current evidence shows that, ciprofloxacin can decrease the mean bacterial load and risk of the exacerbation, and more efficiently eradicate bacteria from sputum in non-cystic fibrosis bronchiectasis patients. Due to limited quality and quantity of the included studies, more studies are required to verify the conclusions.
Cryogels are a type of hydrogel material which are fabricated by cryopolymerization at subzero temperature. Due to their unique macroporous structure, shape memory properties and injectability, cryogels have gained significant interest in the fields of tissue engineering for encouraging the repair and regeneration of injured tissues. In this review, the basic concepts relevant to cryogels are introduced, and then the fabrication principle, the process parameters and the unique properties of cryogel are discussed. Next, the latest advances of cryogels as three-dimensional scaffold for various tissue engineering applications are given. Finally, this review summarizes the current limitations of cryogels, and strategies to further improve their properties for tissue engineering. The purpose of this article is to provide a reference guide for the researchers in related fields.
Objective To investigate the effect of caveolion-1 on the growth and proliferation in human breastcancer MCF7 cell in vitro and in vivo and approach its mechanisms. Methods The plasmid pCI-neo-cav-1 and its corres-ponding empty vector （pCI-neo） were transfected into MCF7 cell line. The expressions of caveolin-1 and vascular endoth-elial growth factor （VEGF） in transfectants were subsequently confirmed by Western blot analysis. A pair of monoclonal cell lines， MCF7/cav-1 and MCF7/vec， were chose for future studies. The colony formation ability of tumor cells was detected by anchorage-independent growth assay. Xenograft tumor models in nude mice were established. Immunohisto-chemistry was used to characterize Ki-67 and VEGF levels in the tumors tissues. Results Caveolin-1 expression was up-regulated in the MCF7/cav-1 cell as compared with the MCF7/vec cell （P＜0.01） and the MCF7 cell （P＜0.01）. Caveolin-1 overexpression markedly reduced the capacity of the cells to form colonies in soft agar （P＜0.01）. Compared with the MCF7/vec cell， VEGF protein expression reduced in the MCF7/cav-1 cell （P＜0.01）. In vivo experiments in nude mice， the overexpression of caveolin-1 resulted in significant growth inhibition of the xenograft breast tumors. The Ki-67 staining was weak and the VEGF staining was negative by immunohistochemistry that indicated the caveolin-1 inhibited the proliferation in human breast cancer MCF7 cell. Conclusion The caveolin-1 might inhibit the growth and proliferation in human breast cancer MCF7 cell through suppressing activation of VEGF signaling pathway.
ObjectiveTo explore the methods of separation， culture， and identification of breast cancer stromal fibroblasts (BCSFs), which could build up a good basis for the further research of function. MethodsBreast cancer tissues were obtained during breast cancer operation， and were cut into pieces with size of 1 mm×1 mm×1 mm under aseptic conditions， then the pieces of the tissues were digested by collagenase Ⅰ and hyaluronidase. Finally the cells separated from the tissues incubated at 37 ℃ with 5% CO2 and 95% air humidified incubator. Morphological characteristics of the fibroblasts were observed under light microscope. The certain proteins were examined by immunohistochemistry （using CK， Vimentin， α-SMA, and TE-7 antibody） and flow cytometric analysis (CD34 and CD45). ResultsThe separated cells begin to attach to the wall of flask within 24 h and reached almost confluency in about 7 d to 10 d . According to identification, the successful rate of separation and culture of BCSFs was 90%（18/20）, and the characteristics of cells showed that morphological characteristics of the fibroblasts was flat spindle， rich cytoplasm, and a flat ovoid cystic nuclear. The fibroblasts in breast cancer tissues showed negative staining for cytokeratin， positive staining for vimentin， alpha-smooth muscle actin, and TE-7， and negative for CD34 and CD45 by flow cytometric analysis. ConclusionsThe fibroblasts in breast cancer tissues could be easily obtained by tissues cuting combined enzyme digestion and rocking technology in vitro. The present study provide an experimental foundation for further studies on fibroblasts in breast cancer.