Long-term serum-free culture of cancer stem/progenitor cells from basal-like breast carcinoma / 中国组织工程研究

BACKGROUND: Breast cancer stem cells, a subgroup of breast cancer cells, have self-renewal and multilineage differentiation potential and are characterized as resistance to chemotherapy, radiotherapy and hypoxia, and high propensity for invasiveness and metastasis, which play an important role in the recurrence and metastasis of breast cancer. The breast cancer stem cell theory can elucidate the pathogenesis of breast cancer to a certain extent, and has great values in the selection of treatment options, the discovery of new therapeutic targets and the prognosis of breast cancer.OBJECTIVE: To isolate the breast cancer stem/progenitor cells from basal-like breast carcinoma and study their self-renewal ability, phenotype and differentiation potential over long-term serum-free suspension culture in vitro. METHODS: Twenty patients with basal-like breast cancer, including 10 patients without any treatment and 10 with neoadjuvant chemotherapy (≥ 4 cycles), were enrolled. A 1 cm3 fresh tumor tissue sample from each patient was taken and immediately sent to the laboratory to mechanically isolate tumor cells. Then, the stem/progenitor cells of basal-like breast carcinoma were enriched in ultra low attachment plates in serum free media as nonadherent mammospheres. Serial sphere formation assay was performed to determine colony formation and self-renewal ability of mammosphere-derived cells. Differentiation was induced by culturing mammosphere-derived cells in DMEM-F12 supplemented with serum but without growth factors. The proportion of CD44+/CD24- and ALDH1+ cell population was evaluated by flow cytometry. RESULTS AND CONCLUSION: The mammospheres formed after inoculation of primary basal-like breast cancer cells isolated from the tumor tissues of breast cancer patients who did not receive neoadjuvant chemotherapy cultured in the serum-free medium with growth factors, while the mammospheres could be directly isolated from the tumor specimens of patients with neoadjuvant chemotherapy. The mammosphere-derived cells could be passaged continuously to form new mammospheres. With the increase of subculture frequency, the number of adherent cells was increased, but the number of new mammospheres decreased, and the spheres became smaller and easier to adhere. The mammosphere-derived cells could be induced to differentiate in the medium supplemented with serum. The CD44+/CD24- and ALDH1+ cells were enriched in mammosphere-derived cells, and these two phenotypic cells were decreased sharply in number or absent after cell differentiation. Most of the mammosphere-derived cells after chemotherapy had stronger potentials of self-renewal and differentiation, with higher proportion of CD44+/CD24- and ALDH1+ cells. In summary, cancer cells with self-renewal and multilineage differentiation potentials exist in the basal-like breast cancer tissues. Neoadjuvant chemotherapy can enrich breast cancer stem/progenitor cells to form spheroid-like cell clusters. Some differences in biological behaviors exist between the stem/progenitor cells from different breast cancer samples, which can vary with environmental factors.

Curcumin hydrogel combined with photodynamics to treat acne / 国际药学研究杂志

Objective To demonstrate the therapeutic effect of curcumin hydrogel combined with photodynamics on acne. Methods Sieving and ultrasound/high pressure homogenization were used to prepare curcumin micropowder.Curcumin-loaded hydro?gel was prepared with Poloxamer 407 and its rheological characteristics were evaluated.The antibacterial activity of curcumin hydrogel irritated with 657 nm laser was assayed by a hole-punching method.Acne model on rabbit ears war established.Fifteen New Zealand white rabbits were randomly divided into five groups:the model group,tretinoin cream group,illumination group,curcumin hydrogel group,and curcumin hydrogel/illumination group.The therapeutic effects on acne were evaluated by the morphological and histopatho?logical examination.Results The micropowdered curcumin prepared with ultrasound/high pressure homogenization had smaller size and more uniform distribution(size range:5-20 μm.)than the sieving-prepared curcumin micro-powders(10-45 μm).The prepared hydrogels being a pseudoplastic fluid had the properties of gelatin.The antibacterial effect of curcumin hydrogel/illumination was bet?ter than that of curcumin hydrogel.The inhibition zone diameters were(9.4±0.92)and(12.57±1.12)mm for the curcumin hydrogel and the curcumin hadrogel/illumination groups,respectively.The therapeutic effect for the treatment of acne on the rabbit ears with ac?ne was better in the curcumin hydrogel/illumination group than in both the illumination and the curcumin hydrogel groups. Conclu?sion Curcumin hydrogel combined with photodynamic therapy showed better therapeutic efficacy on acne treatment.It provides an ideal choice for acne treatment.

Curcumin temperature-sensitive in situ hydrogels for treatment of vaginal candidiasis / 国际药学研究杂志

Objective To investigate the therapeutic effect of curcumin in situ hydrogels for treatment of vaginal candidiasis. Methods In situ hydrogels were prepared using poloxamer 407 and 188 as the gel matrix.Gelation temperature,gelation time and rheological characteristics were determined.Mice models with vaginal candidiasis were established.The animals were divided into six groups:the healthy mice group,the model group,the positive control group(ketoconazole cream),curcumin-loaded in situ hydrogels of 0.025,0.10 and 0.20 g/ml.Before treatment and after the fourth and eighth treatments,the vaginal smears were collected and Gram-stained.The vagina washings fluid of mice were collected to determine DNA of Lactobacillus.On 8thday,the mice were sacrificed and the vaginal tissues were excised for pathological detection.Results The gelation temperature and time of in situ hydrogels were 29℃and 2′30″,respectively.Rheological profiles demonstrated that it showed good retention effect in vivo.Compared with the model group, curcumin hydrogels of 0.025,0.10 and 0.20 g/ml could inhibit the growth of Candida albicans,inhibit vaginal mucosal edema,inflam?matory hyperplasia,and decrease the expression of inflammatory factors including TNF-α(P<0.01)and IL-1β(P<0.01).Howev?er,curcumin hydrogels could not influence the growth of lactobacillus in the vagina.It indicated that curcumin hydrogels did not de?pend on lactobacillus to treat fungal vaginitis.Conclusion Curcumin in situ hydrogels could inhibit candidiasis growth,decrease ede?ma of vaginal mucosa and inflammatory hyperplasia,and reduce the expression of inflammatory factors.It provides a novel choice for the treatment of clinical vaginal candidiasis.

Study on function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang (XFZYT).</p><p><b>METHOD</b>Serum pharmacology technique was adopted to treat endothelial cells ECV304 with XFZYD containing serum and blank serum with concentrations of 1.25%, 2.5% and 5%. Methyl thiazolyl tetrazolium (MTT) assay, boyden chamber migration assay and in vitro vessel formation assay were used to test endothelial cell proliferation, migration and angiogenesis capacities. ELISA, immunohistochemistry and RT-PCR were used to detect secretion and expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2).</p><p><b>RESULT</b>XFZYT-CS with a concentration of 1.25% only affected angiogenesis capacity in vitro. XFZYT-CS with a concentration of 2. 5% promoted cell proliferation, migration and angiogenesis capacities and significantly increased VEGF level and VEGF/VEGFR2 expressions. XFZYT-CS with a concentration of 5% only up-regulated intracellular VEGF expression and thereby improving endothelial cell proliferation, migration and angiogenesis.</p><p><b>CONCLUSION</b>XFZYT can induce endothelial cell proliferation, migration and angiogenesis by up-regulating VEGF-VEGFR2 pathway, which partially proves its angiogenesis effects.</p>

A microarray analysis of angiogenesis modulation effect of Xuefu Zhuyu Decoction on endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the angiogenesis modulation mechanism of Xuefu Zhuyu Decoction () on the endothelial cell line ECV304.</p><p><b>METHODS</b>ECV304 cells were treated with 2.5% Xuefu Zhuyu Decoction-containing serum (XFZYD-CS) for 24 h, 48 h or 72 h. Thiazolyl blue tetrazolium bromide (MTT), fluorescence activating cell sorter (FACS), migration, adhesion and in vitro tube formation assays were conducted to confirm an angiogenesis effect of XFZYD at 3 time points. An analysis of angiogenesis regulator profiles was performed at 3 times with real-time polymerase chain reaction (RT-PCR) superarray.</p><p><b>RESULTS</b>At 48 h, XFZYD-CS induced ECV304 significantly improved cell viability, number in S phase, migration, adhesion and tube formation. At 24 h and 72 h, only cell migration was elevated. Microarray results showed that the expression of 27 angiogenesis-related genes was changed.</p><p><b>CONCLUSION</b>XFZYD-CS treatment induced angiogenesis on ECV304 cells with significant cellcular changes occurring at 48 h and genetic changes as early as 24 h.</p>

Analysis of variation trends of thyroid cancer treated in Tianjin Cancer Hospital form 1954 to 2009 / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate and analyze the variation trends in the pathological composition of thyroid cancer patients treated in Tianjin Cancer Hospital from 1954 to 2009.</p><p><b>METHODS</b>To retrospectively analyze the incidence and clinical features of different pathological types of thyroid cancers in 4342 patients between different time periods from 1954 to 2009.</p><p><b>RESULTS</b>In the four main pathological types of thyroid cancers, the component ratio of papillary thyroid cancer in every period was 68.1%, 78.3%, 81.3%, 82.1%, 85.8%, respectively, while the morbidity of patients with papillary thyroid carcinoma concurrent with Hashimoto's thyroiditis was increased, so was the proportion of tumors in diameter < or = 2 cm. The proportion of follicular thyroid carcinoma and anaplastic thyroid carcinoma was decreasing accordingly; however, the proportion of medullary thyroid carcinoma did not change significantly.</p><p><b>CONCLUSIONS</b>The pathological classification of the thyroid carcinoma patients has significant changes in the 4342 cases treated in our Hospital from 1954 to 2009. The proportion of papillary carcinoma is increased, while that of follicular carcinoma and anaplastic carcinoma is decreased. The reasons might attribute to the improved level of consultations and iodized diet or other factors.</p>

Effect of xuefu zhuyu decoction in inducing angiogenesis gene regulation of endothelial cell line ECV304 / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the acting mechanism of endothelial cell line ECV304 in regulating angiogenesis induced by Xuefu Zhuyu Decoction (XFZY).</p><p><b>METHODS</b>The angiogenesis effect of XFZY-contained serum (XFZY-CS) was confirmed by observing its impact on proliferation, cell cycle, migration of ECV304 and on vascular neogenesis in vitro. Then the effect of XFZY on various angiogenesis controlling factors was analyzed with gene chip microarray technique.</p><p><b>RESULTS</b>Treatment of XFZY-CS in 2.5% concentration for 48 h showed evident actions of enhancing ECV304 activity, increasing cell numbers of S phase, inducing cell migration and promoting the in vitro angiogenesis. Meanwhile, expressions of four angiogenesis controlling genes were up-regulated and 10 were down-regulated.</p><p><b>CONCLUSION</b>The angiogenesis mechanism of ECV304 induced by XFZY is complex, it shows a multi-pathway and multi-target feature.</p>

The effect of Xuefu Zhuyu Decoction on in vitro endothelial progenitor cell tube formation / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of Xuefu Zhuyu Decoction ()-containing serum (XFZYD-CS) on endothelial progenitor cell (EPC) tube formation in vitro.</p><p><b>METHODS</b>Mononuclear cells from rat bone marrow were prepared in a Ficoll density gradient centrifuge. EPCs were separated by the differential attachment method, and observed with inverted microscope for the effect of XFZYD-CS on EPC tube formation.</p><p><b>RESULTS</b>After one day, EPCs exposed to the serum containing 5%, 10% and 15% XFZYD-CS formed typical tubes or vessel networks. The tube formation time was two days ahead of the control group and the size of most tubes in the serum groups was smaller than in the control group.</p><p><b>CONCLUSION</b>XFZYD-CS could induce EPC angiogenesis and hasten tube formation, especially in capillary vessels. The study provides experimental evidence for the plausibility of Xuefu Zhuyu Decoction in the treatment of ischemic diseases.</p>