[Effect of sodium alginate-g-deferoxamine/chitosan microspheres on osteogenic differentiation of rat bone mesenchymal stem cells].

PURPOSE: To explore the effect of sodium alginate-g-deferoxamine/chitosan (SA-g-DFO/CS) microspheres on proliferation and osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs). METHODS: A kind of SA-g-DFO/CS microsphere was developed through electrostatic interaction between porous chitosan microspheres and sodium alginate chemically grafted on the surface of DFO. Its morphology, porosity rate, pore size and sustained release of DFO in vitro were examined. Rat BMSCs were isolated and co-cultured with microspheres in osteogenic differentiation medium. MTT assay was used to study the influence of cell proliferation, and Calcein-AM/PI staining was used to observe the cell viability. Alkaline phosphatase (ALP) activity assay was conducted. PCR was used to detect the expression of genes related to angiogenesis and osteogenesis. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: The SA-g-DFO/CS porous microspheres were successfully prepared with a sustained re6lease of DFO. Compared with SA/CS microspheres, the SA-g-DFO/CS microspheres were conducive to cell proliferation and differentiation, with the increases in expression level of ALP, related angiogenesis genes HIF-1α, VEGF and osteogenesis genes COLI, OCN. CONCLUSIONS: The SA-g-DFO/CS porous microspheres can provide a new choice for the development of alveolar bone regeneration.

A new antiviral pregnane from a gorgonian-derived Cladosporium sp. fungus.

A new pregnane, 3α-hydroxy-7-ene-6,20-dione (1), and five known steroids (2-6), along with one known steroidal glycoside (7) were obtained from the fungus Cladosporium sp. WZ-2008-0042 cultured from a gorgonian Dichotella gemmacea collected from the South China Sea. The structure and absolute configuration of the new compound (1) were elucidated by comprehensive spectroscopic data and X-ray diffraction data. The compound has a rare configuration of 3α-OH that is different from most of pregnanes. All of the isolated compounds were evaluated for their antiviral activities against respiratory syncytial virus (RSV). Among them, 1 exhibited potential antiviral activity with the IC50 value of 0.12 mM.

Survey on physicians who engage in cancer diagnosis and treatment from third-grade class A hospitals regarding the perceptions of cancer patients' fertility preservation / 上海交通大学学报（医学版）

Objective · To explore the perceptions and current practice barriers of Chinese physicians who engage in cancer diagnosis and treatment from third-grade Class A hospitals regarding cancer patients' perceptions of fertility preservation (FP).Methods · A study was conducted in physicians from 4 third-grade Class A hospitals with different clinical specialties assisting cancer patients through a structured self-report questionnaire between January 2017 and December 2017.A total of 179 medical oncologists,77 radiation oncologists and 79 surgeons were included.Their information on gender,age,title,education background and perceptions of FP was obtained.Logistic regression analysis was used to examine the correlation between the recommendation of FP and risk factors.Results· There were 335 physicians,including 88 male physicians and 247 female physicians,with an average age of (35.94±6.27) years (range from 23-59 years) in the current study.Although 96.4％ of the physicians knew that chemotherapy and radiation had a profound effect on impairment of fertility and 85.1％ of them thought it was necessary to recommend FP,only 28.1％ of them gave FP-related recommendations to the cancer patients.The oncologists and surgeons,female physicians,and those with higher professional titles and education background were more likely to make the FP recommendation.Among 63.3％ of the physicians knowing male FP,only 37.9％ and 21.2％ of them noted the exact methods and place for FP,respectively.Similarly,for the 65.1％ of the physicians knowing female FP,the percentage was 49.9％ and 24.5％,respectively.When it came to the barriers of FP decision-making,32.8％ of the oncologists reported their concerns on whether cancer patients were suitable to reproduce.Secondly,the physicians honestly admitted that they lacked expertise in FP and worried about that FP would delay cancer treatment.Conclusion· Physicians who engage in cancer diagnosis and treatment lack the awareness and knowledge background of FP recommendation for cancer patients.It is important to improve the perceptions of cancer patients' FP through standardize training.

Increased expressions of vascular endothelial growth factor (VEGF), VEGF-C and VEGF receptor-3 in prostate cancer tissue are associated with tumor progression.

AIM: To investigate the differences in microvessel densities (MVD) and the expressions of vascular endothelial growth factor (VEGF), VEGF-C and VEGF receptor-3 (VEGFR-3) between prostate cancer (PCa) tissues and adjacent benign tissues, and to explore the correlations among MVD, Jewett-Whitmore staging, Gleason scores and expressions of VEGF, VEGF-C and VEGFR-3 in the progression of PCa. METHODS: An immunohistochemical approach was adopted to detect the expressions of CD34, VEGF, VEGF-C and VEGFR-3 in both cancer areas and peripheral benign areas of 71 primary prostatic adenocarcinoma specimens. A statistic analysis was then performed according to the experimental and clinic data. RESULTS: Significantly upregulated expressions of VEGF, VEGF-C and VEGFR-3 were all found in malignant epithelium/cancer cells compared with adjacent benign epithelium (P<0.01). Patients in stage D had a significantly higher score than patients in stage A, B or C when comparing the expression of VEGF-C or VEGFR-3 in the tumor area (P<0.01). In addition, significant correlations were observed between Jewett-Whitmore staging and VEGF-C (r(s)=0.738, P<0.01), clinical staging and VEGFR-3 (r(s)=0.410, P<0.01), VEGF-C and Gleason scores (r(s)=0.401, P<0.01), VEGFR-3 and Gleason scores (r(s)=0.581, P<0.001) and MVD and VEGF (r(s)=0.492, P<0.001). CONCLUSION: Increased expressions of VEGF and VEGF-C were closely associated with progression of PCa. The main contribution of increased VEGF expression for PCa progression was to upregulate MVD, which maintained the growth advantage of tumor tissue. However, the chief role of increased expressions of VEGF-C and VEGFR-3 was to enhance lymphangiogenesis and provide a main pathway for cancer cells to disseminate.

Study on the bone marrow mesenchymal stem cells induced drug resistance in the U937 cells and its mechanism / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The hematopoietic microenvironment (HM) plays a critical role in malignant cell growth, patient survival, and response to chemotherapy in hematologic malignancies. However, mechanisms associated with this environmental influence remain unclear. In this study, we investigated the role of bone marrow derived mesenchymal stem cells (MSCs) in U937 cell line, to find out the relations between leukemia drug resistance and the MSCs.</p><p><b>METHODS</b>U937 cells were cultured in suspension or grew adherently with MSCs. The cell growth curve was drawn and the cell cycle was measured by flow cytometry. Apoptosis and sensitivity of U937 to daunoblastina (DNR) were quantified by DNA ladder detection and trypan blue exclusion assays, respectively. The gene expression profile chip technology was used to determine and analyze the changes in apoptosis-related gene expression after adherent culture and the expression of MDR1 mRNA was assessed by reverse transcriptional polymerase chain reaction (RT-PCR) at the same time.</p><p><b>RESULTS</b>In the adherent culture, the proliferation of the U937 cells was inhibited, the G0/G1 phase cells increased (F = 64.9726, P < 0.0001), G2/M phase cells were decreased (F = 98.1361, P < 0.0001) and the natural apoptosis rate was decreased (F = 24.0866, P < 0.0001) compared with those in the suspended culture. U937 cell viability was enhanced and cell apoptosis was blocked during DNR treatment in adherent culture with MSCs. Thirty-nine differently expressed genes were screened from the 487 apoptosis related genes in the adherent culture U937 cells. Among the 37 upregulated genes, Bcl-XL was upregulated most significantly. Two genes were downregulated. Adherent culture did not induce MDR1 mRNA expression in U937 cells.</p><p><b>CONCLUSIONS</b>MSCs play a role in modulating the proliferation of U937 cells and response of U937 cells to DNR, and Bcl-XL apoptosis-inhibiting gene may be most important in determining the sensitivity of leukemic cells to treatment, which is not related to MDR1.</p>

Influence of human mesenchymal stem cells on cell proliferation and chemo-sensitivity of K562 cells / 中国实验血液学杂志

This study was aimed to compare K562 cell proliferation, chemo-sensitivity and alteration of MDR1 before and after adhesive culture with MSC, so as to evaluate the relationship between chemodrug-resistance of leukemia cells and hemopoietic microenvironment. K562 cell cultivated in suspension and adhesively cultivated with MSC were collected respectively and cell proliferation curves were drawn; the cell cycle was determined by flow cytometry; the effect of chemotherapy on cellular viability and apoptosis of K562 cell was investigated, the MDR1 gene expression was determined by RT-PCR. The results showed that K562 cells adhesively cultivated with MSC were inhibited and cells in G0/G1 increased (P < 0.05), cells in S phase decreased (P < 0.05) and those in G0/G1 increased (P < 0.01), compared with that cultivated in suspension. In process of daunomycin-inducing apoptosis, K562 cell apoptosis in the adhesive culture with MSC was inhibited (P < 0.05). MDR1 gene expression in K562 cells was not induced or altered by adhesive co-cultivation. It is concluded that by co-culture of cell-cell contact with MSC, growth suppression and induction of chemo-resistance of K562 cells take place. The mechanism, however, seems not relevant with MDR1.

The relationship between abnormal MDR gene expression and chemotherapy response in lymphoid malignancies / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of multidrug resistance-1 (MDR(1)), Topoisomerase II (Topo II), glucocorticoid receptor (GCR) and their correlation with relapse rate and chemotherapy response in lymphoid malignancies.</p><p><b>METHODS</b>The expression of MDR(1), Topo II and GCR in 189 patients with lymphoid neoplasms was examined by RT-PCR, slot blot and ligand-labelled methods.</p><p><b>RESULTS</b>(1) The expressions of MDR(1), Topo II, GCR in untreated and relapsed/refractory patients with ALL, NHL, NHL-L, MM were significantly abnormal at varying levels, especially in the relapsed/refractory group. (2) The complete remission (CR) rate of MDR(1) high expression group (MDR(1)(+)) was significantly lower than that of MDR(1) negative expression (MDR(1)(-)) group (P < 0.05), and the relapse rate of MDR(1)(+) group was significantly higher than that of MDR(1)(-) group (P < 0.05). In untreated patients, the relapse rate in the Topo II low expression (Topo II(-)) group was positively higher than Topo II high expression (Topo II(+)) group (P < 0.05), whereas in the relapsed/refractory patients, the CR rate of Topo II(-) group was significantly lower than that of Topo II(+) group (P < 0.05). In the untreated and relapsed/refractory patients, the CR rates of low GCR expression (GCR(-)) group was obviously lower than that in the normal GCR expression group (P < 0.05). (3) Considering mono-drug resistance mechanism, CR rate of MDR(1)(+) group was the lowest, Topo II(-) group took the second place and GCR(-) group was the highest. As multiple drug resistance mechanisms coexisted, the CR rate of MDR(1)(+) + Topo II(-) + GCR(-) group and MDR(1)(+) + Topo II(-) group (11.1% and 15.4%, respectively) were significantly lower than that of MDR(1)(+), Topo II(-) and GCR(-) group (36.7%, 48.0% and 53.8%, respectively; P < 0.05 - P < 0.001).</p><p><b>CONCLUSION</b>There are primary and acquired drug resistance in lymphoid neoplasms. The high expression of MDR(1), low expression of Topo II and GCR are positively related to low chemotherapy response rate and high 2-year relapse rate. Co-analysis of MDR(1), Topo II and GCR may play an important role on chemotherapy response and prognostic judgment in lymphoid neoplasms.</p>

Study of apoptosis gene expression in U937 cells induced by adhesion culture with mesenchymal stem cell / 中华血液学杂志

<p><b>OBJECTIVE</b>To compare apoptosis gene expression profiling of U937 cells in suspension culture with that cultivated with mesenchymal stem cells (MSCs), and find out the relationship between drug resistance of leukemia cells and hemopoietic microenvironment.</p><p><b>METHODS</b>U937 cells were cultivated in adhesion culture with MSCs and in suspension culture for 48 hours. Cell cycle was determined by flow cytometry and gene expression profiling by cDNA microarray.</p><p><b>RESULTS</b>Compared with that in suspension, G(0)/G(1) fraction of U937 cells increased in adhesion culture (45.3 +/- 3.1)% vs (32.6 +/- 2.1)%, respectively (P < 0.05), whereas G(2)/M fraction and apoptosis rate were decreased. After 48 h twenty-eight differential expression genes were screened out in 487 apoptosis-related genes, among which 27 were up-regulated and were mainly apoptosis-suppressor genes, apoptosis-promoter genes, cell cycle positive control genes and cell cycle negative control genes. But Bcl-XL was up-regulated most obviously. The only one gene down-regulated was an apoptosis promoter gene.</p><p><b>CONCLUSION</b>Adhesion culture with MSCs can lead to growth suppression and decrease natural apoptosis of U937 cells. The mechanism was multiple gene effects, but Bcl-XL may be of the most importance.</p>

The endocrine and reproductive function of the female Yucheng adolescents prenatally exposed to PCBs/PCDFs.

BACKGROUND: Polychlorinated biphenyls (PCBs) and related compounds such as polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-dioxins (PCDDs) alter sexual maturation and endocrine function in animals. In 1978-1979, a mass poisoning occurred in central Taiwan from cooking oil contaminated by heat-degraded PCBs and oxidated compounds PCDFs. We tested the hypothesis that in utero exposed to PCBs/PCDFs alter sexual maturation, endocrine, and reproductive function in the human pubescent females. METHODS: In 1997-1999, girls aged 13-19 years, born to mothers exposed to PCBs/PCDFs, was invited to participate in the study. Menstruation characteristic was recorded daily for 84 days and serum levels of estradiol, LH, FSH, and testosterone were measured on the 3rd day of menstruation. RESULTS: A total of 17 exposed girls and controls participated, the exposed girls reported shorter mean duration of bleeding per cycle than 16 unexposed (5.5 vs 6.5 days, P=0.0055). There was a higher rate of irregular menstrual cycle in the exposed girls (40% vs 0%, P=0.018). Serum levels of estradiol (P=0.016) and FSH (P=0.061) were higher in exposed girls as compared to controls. CONCLUSIONS: We conclude that prenatal exposure to PCBs/PCDFs resulted in abnormal menstruation and higher estradiol and FSH levels in follicular phase of menstrual cycle in adolescent girls.

A cohort study of behavioral problems and intelligence in children with high prenatal polychlorinated biphenyl exposure.

BACKGROUND: In 1978, about 2000 persons in Taiwan were poisoned when their cooking oil was contaminated during manufacture with heat-degraded polychlorinated biphenyls, which are toxic, very widespread pollutant chemicals. The chemicals cannot be metabolized or excreted, and 8 of the first 39 children born to affected women died. When examined in 1985, 117 surviving children were found to have ectodermal defects, developmental delay, and disordered behavior. We have continued to observe the children. METHODS: From 1992 through 1995, 118 children born between 1978 and 1985 (during or after their mothers' exposure) and 118 matched neighborhood control children had cognitive function measured yearly with the Wechsler Intelligence Scale for Children-Revised and behavioral problems measured with the Achenbach Child Behavior Checklist and the Rutter Child Behavior Scale A. RESULTS: The exposed children scored 3 points (P =.05) lower than control children for IQ; 3 points (P =.002) higher on the Child Behavior Checklist (an effect size similar to the sex difference); and 6 points (P<.001) higher on the Rutter scale (3 times the sex difference). Birth year x exposure interactions, testing whether children born long after the exposure were as affected as those born soon after, were small and not significant. Age x exposure interactions, testing whether the children improved relative to control children as they got older, were significant only for the Rutter scale. CONCLUSIONS: Prenatal exposure to these compounds produces long-lasting cognitive and behavioral damage, but there is some evidence of recovery.