BACKGROUND

Vitamin D (VD) deficiency results in a worse prognosis in patients with chronic lymphocytic leukemia (CLL) and may affect the production of cytokines. Nonetheless, there is the lack of studies dealing with VD supplementation and its impact on chemokines in CLL patients.

OBJECTIVE

The primary endpoint of our interventional study was to evaluate the effect of cholecalciferol supplementation on serum chemokines levels in CLL patients.

METHODS

Eighteen subjects with CLL were enrolled for the study. Six-month-long cholecalciferol supplementation was performed in CLL patients with serum 25-OH-<em>D</em>3 levels below 30 ng/ml. Cytokines levels were assessed at the beginning of the study and after 6 months. Baseline measurements of cytokines were compared to those in apparently healthy controls.

RESULTS

Increased levels of CCL2, CCL3, CCL4, CXCL8, CXCL10, TNFα, bFGF, G-CSF, and VEGF were found in CLL patients in comparison with the healthy controls. In the course of the VD supplementation a decrease in serum levels of chemokines CCL11, CCL3, and cytokine PDGF-BB was observed. The decrease of CCL11 was found in CLL patients on VD supplementation solely, whereas the decrease of CCL3 and PDGF-BB was observed in CLL subjects on both chemotherapy and VD supplementation.

CONCLUSIONS

The VD supplementation may exert beneficial effect on chemokines levels in CLL patients with VD deficiency.

OBJECTIVE

To investigate effects of low-dose cyclophosphamide and prednisone (CP) metronomic chemotherapy on microvessel density of bone marrow, serum vascular endothelial growth factor (VEGF) and platelet derived growth factor BB (PDGF-BB)in multiple myeloma (MM) patients.

METHODS

54 refractory or relapsed MM patients were treated with CP metronomic chemotherapy consisted of oral cyclophosphamide (CTX, 50 mg/d) and prednisone (Pred, 15 mg/d). Bone marrow and peripheral blood of each patient were collected before and 2, 4, 6 months after treatment. Among the 37 assessable patients, 30 cases were responsive with the response rate of 81.08%. Another 17 cases were follow-uped less than 6 months or failure to obtain serum samples or lost to follow-up. Microvessel density of bone marrow was measured by immunohistochemistry and serum VEGF/PDGF-BB expression was analyzed by ELISA in the 37 assessable patients.

RESULTS

2, 4, 6 months following CP metronomic chemotherapy, microvessel densities of bone marrow in the responders were 33.1 ± 4.8/HP, 24.8 ± 3.7/HP, 19.7 ± 2.1/HP respectively; the expressions of VEGF were (394 ± 57) ng/L, (268 ± 32) ng/L and (217 ± 20) ng/L respectively; the expressions of PDGF-BB were (304 ± 31) ng/L, (274 ± 31) ng/L and (196 ± 22) ng/L respectively. After CP metronomic chemotherapy, there were significantly lower of microvessel density, VEGF and PDGF-BB levels than pretreatment \[MVD 48.5 ± 5.9/HP, VEGF (517 ± 60) ng/L, PDGF-BB (484 ± 60) ng/L\]in the responders (P < 0.01). While in the non-responders, after treated by CP metronomic chemotherapy for 2 months, microvessel density, the expression of VEGF and the expression of PDGF-BB were 32.5 ± 4.7/HP, 512 ± 39 ng/L and (452 ± 39) ng/L respectively. There were no significant changes of MVD, VEGF and PDGF-BB levels compared with pretreatment \[MVD 33.2 ± 5.6/HP,VEGF (498 ± 55) ng/L, PDGF-BB (488 ± 44) ng/L\] (P>> 0.05).

CONCLUSIONS

Our findings suggested that continuous low-dose CP metronomic chemotherapy could decrease microvessel density of bone marrow in MM patients. Furthermore, it down-regulated expression of serum VEGF and PDGF-BB to exert its anti-angiogenesis in MM.

This study sought to assess the effects of Quercetin and Enalapril on urinary protein excretion and amount of platelet-derived growth factor-B (PDGF-B) and vascular endothelial growth factor-1 (VEGF-1) in renal tissue of diabetic rats. 29 streptozotocin-induced diabetic rats were divided into 3 groups, diabetic control group (group D, n=12); Enalapril group (group E, n=10); Quercetin group (group Q, n=7). In addition, there was one normal control group (group N, n = 5). The urinary protein excretion of 24 hours was measured at 4, 8, 12 weeks. All rats were sacrificed at 12 weeks. The amounts of PDGF-B and VEGF-1 in renal tissue were measured by immunohistochemical techniques. The 24-h levels of urinary protein excretion of D,Q and E groups were higher than that of N group; the level of Q, E groups were lower than that of D group; there was no difference between Q and E group. The expression levels of PDGF-B and VEGF-1 in renal tissue of D, Q, and E groups were significantly higher than that of N group the levels of Q and E groups were significantly lower than that of D group, no difference was found between Q and E group. The protective role of Quercetin and Enalapril in lowering urinary protein excretion may be related to the decreased amounts of PDGF-B and VEGF-1 in renal tissue.

TRC105 is an anti-endoglin antibody currently being tested in combination with VEGF inhibitors. In the phase Ib trial, 38 patients were treated with both TRC105 and bevacizumab (BEV), and improved clinical outcomes were observed, despite the fact that 30 patients (79%) were refractory to prior anti-VEGF therapy. Plasma samples were tested for angiogenic and inflammatory biomarkers at baseline and on-treatment. To provide broader context of this combination biomarker study, direct cross-study comparisons were made to biomarker studies previously conducted in patients treated with either BEV or TRC105 monotherapy. Upon treatment with BEV and TRC105, pharmacodynamic changes in response to both BEV (PlGF increase) and TRC105 (soluble endoglin increase) were noted. In addition, distinct patterns of change were identified (similar, opposing, neutralizing). Similar patterns were observed when the combination elicited similar effects to those observed with monotherapy treatment (i.e., decreases of Ang-2, increases of IL6 and VCAM-1). Opposing patterns were observed when the combination led to opposing effects compared with monotherapy treatment (i.e., TGFβ1, PDGF-AA and PDGF-BB, PAI-1). Lastly, neutralizing patterns were observed when one drug led to increase, whereas the other drug led to decrease, and the combination elicited no overall effect on the marker (i.e., VEGF-A, VEGF-D, and IGFBP-3). Patients achieving partial responses or stable disease from the combination exhibited significantly lower expression of E-Cadherin, HGF, ICAM-1, and TSP-2 at baseline. Taken together, the novel biomarker modulations identified may deepen our understanding of the underlying biology in patients treated with BEV and TRC105 compared with either drug alone. Mol Cancer Ther; 17(10); 2248-56. ©2018 AACR.

OBJECTIVE

The growth factor midkine (MK) is a protein that is involved in cancer, inflammation, immunity. Vitamin D is a potent immunomodulator. Anti-Saccharomyces cerevisiae antibody (ASCA) is reported in autoimmune disorders, some of which are among the causes of vitamin D deficiency. The objective of this study was to investigate a possible association of MK and ASCA with vitamin D deficiency.

METHODS

208 adults presented to internal medicine outpatient clinic for history and physical examination has been studied. Serum biochemistry, vitamin D, MK, ASCA-IgG and -IgA, IL-1β, IL-6, IL-8, TNF-α, PDGF, VEGF were obtained.

RESULTS

Vitamin D deficiency was 74.2%. Serum MK level was significantly higher in vitamin D-deficient compared to vitamin D-sufficient individuals (1138.1 ± 262.8 vs 958.6 ± 189 pg/mL, respectively; P < 0.009). Serum MK levels were also significantly higher in both ASCA-IgG and -IgA positives compared to negatives (1318.5 ± 160.3 vs 1065.5 ± 256.1, P = 0.008 and 1347.7 ± 229.7 vs 1070.1 ± 250.9 pg/mL, P = 0.011, respectively). Vitamin D was significantly lower in ASCA positives (P = 0.044).Vitamin D showed positive correlation with IL-1β (r 0.338, P < 0.009) and negative correlation with VEGF (r -0.366, P < 0.004).

CONCLUSIONS

MK was significantly elevated in vitamin D deficiency and associated with ASCA positivity which was significantly increased in vitamin D deficiency. These findings suggested that molecular mechanism of vitamin D deficiency may be related with some inflammatory processes.

OBJECTIVE

This study was to investigate whether prestorage leukoreduction could decrease the accumulative concentration of tumor-associated cytokines in supernatant of stored packed red blood cells (pRBC) and to study the effect of prestorage leukoreduction on proliferation of HepG2 tumor cells by in vitro. The leukoreduced (LR) and non-leukoreduced (NLR) pRBC were equally obtained from one donation and were stored under 2 °C-6°C. The supernatants of pRBC in these two group were performed by centrifugation with 1 006×g for 10 min at day 0 and 35 d. The enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of normal T cells and secretory factor (RANTES/CCL5), as well as the accumulative concentrations of tumor-necrosis factor (TNF-α), platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF) and monocyte chemotactic protein-1 (MCP-1) in pRBC supernantant of above-mentioned two groups. After HepG2 cells was cultured with the supernatant of NLR-pRBC and LR-pRBC at the end of day 35 together for 48 hours, the methyl thiazolil tetracolium (MTT) method was used to measure the proliferation of tumor cells in vitro.

RESULTS

The accumulative concentration of 5 cytokines in supernatants of above menthioned two groups increased in different degrees along with the prolongation of storage time,that is, the accumulative concentrations of 5 cytokines at 35 d were higher than that at day 0, in which the change of VEGF accumu-lative concentration showed statistical significance, its accumulative concentration in NLR group at day 35 elevated to 549.61 ± 299.43 pg/ml, and was higher than that in LR group (95.46 ± 110.87 pg/ml) (P < 0.05). The experiment of HepG2 cell proliferation indicated that the supernatant of LR pRBC group produced less proliferation of tumor cells with OD value 0.40 (95% CI, 0.38-0.42) than that of NLR pRBC group with OD value 0.49 (95% CI, 0.43-0.55) (P < 0.05).

CONCLUSIONS

The prestorage leukoreduction has been confirmed to decrease the accumulative level of cytokines, particalarly decrease the accumulative level of VEGF, moreover, it may be a factor for inhibiting the proliferation of tumor cells in vitro.

The human parathyroid hormone N-terminal fragment [hPTH-(1-34)] increases the conversion of exogenous unsaturated fatty acids to prostaglandins (PGs) in calvarial homogenates. Enzyme activities were completely blocked by indomethacin (5 x 10(-7) M), a PG synthase inhibitor, and actinomycin D (5 muM), an inhibitor of transcription, by binding to DNA. In addition, a potent inhibitor of protein synthesis, cycloheximide (10 muM), totally inhibited the stimulating effect of hPTH-(1-34) on prostaglandin endoperoxide synthase (PG synthase, EC 1.14.99.1). The stimulatory effect of hPTH-(1-34) on PG synthase was also reduced by the addition of stannous chloride. However, epidermal growth factor (EGF), platelet-derived activating factor (PDGF), and ionophore A23187 did not show the same stimulating effect as hPTH-(1-34) on PG synthase in calvaria. The results further demonstrated that PG synthase is a membrane-bound enzyme in chick calvaria. In this communication, evidence is presented that hPTH-(1-34) stimulates the de novo synthesis of PG synthase as demonstrated by the increased activity in calvarial homogenates and microsomes.

OBJECTIVE

To explore the role of the basic fibroblast growth factor (bFGF) in the directional differentiation of bone marrow mesenchymal stem cells (BMSCs) into Leydig cells.

METHODS

After purification and identification, we inoculated the third-generation BMSCs of SD rats onto a six-orifice board and then randomly divided them into groups A (normal saline control), B (human chorionic gonadotropin [hCG] + platelet-derived growth factor [PDGF] induction), C (hCG + PDGF + 5.0 ng/ml bFGF induction), D (hCG + PDGF + 10.0 ng/ml bFGF induction), and E (hCG + PDGF + 20.0 ng/ml bFGF induction). On the 7th, 14th and 21st day of induction, we observed the morphological changes of the cells and measured the level of testosterone (T) and expression of 3 beta hydroxy steroid dehydrogenase (3β-HSD) in the supernatant by immunofluorescence staining.

RESULTS

After induction, the BMSCs of groups B, C, D, and E exhibited microscopic features of enlarged size, inter-connection, long-shuttle or irregular shape, adherent growth, and large round nuclei, all characteristic of Leydig cells. With the prolonging of time and enhanced concentration of bFGF, gradual increases were observed in the T level and the count of 3β-HSD-positive BMSCs in the four induction groups, with statistically significant differences between group B and groups C, D, and E (P < 0.05), as well as between group C and groups D and E (P < 0.05), but not between D and E (P>> 0.05).

CONCLUSIONS

The bFGF has an obvious promoting effect in the in vitro induced differentiation of rat BMSCs into Leydig cells.

We investigated the relationship between the motility of hepatic Ito cells and their myofibroblastic transformation in cultures. Ito cells were freshly isolated from rat liver and cultured in a 10% FBS-supplemented medium. On day 2 after beginning the culture, transmission electron microscopy and oil red O staining showed that Ito cells possessed numerous lipid droplets but not actin filaments in the cytoplasm. On day 8, actin filaments were abundantly found in the cytoplasm, whereas lipid droplets dramatically decreased in number. Western blot analysis also demonstrated that protein levels of alpha-smooth muscle actin in the cell markedly increased with time, but no obvious change was detected in those of desmin and tubulin beta. In Boyden's chamber assay, the migration of Ito cells, which was induced by platelet-derived growth factor-BB (PDGF-BB), was activated in a time-dependent manner. This migration of transformed Ito cells was independent of the degree of their adhesion to various substances of the extracellular matrix. Among these molecules, laminin showed the highest effect upon the migratory activity. The migration of transformed Ito cells on laminin was completely inhibited by cytochalasin D, colchicine, or taxol. Furthermore, their adhesion to the matrix was also decreased by cytochalasin D or colchicine, but not by taxol. These findings indicate that the motility of Ito cells is acquired in conjunction with their myofibroblastic transformation, which is accompanied by morphological changes with a new organization of actin filaments. The results also suggest that microtubules as well as the extracellular matrix are deeply associated with the motility of Ito cells.

BACKGROUND

The sequential appearance of cyclins D and E is thought to initiate subsequent DNA synthesis in proliferating cells. Previous studies have reported that DNA synthesis in cultured rat vascular smooth muscle cells (VSMCs) was suppressed by the HMG-CoA reductase inhibitor lovastatin. The effects of lovastatin on cell cycle regulatory proteins in proliferating VSMCs, however, are largely unknown. Thus, we investigated the sequential expression of cyclin DDK) 4, CDK2, and p27Kip1 in cultured rat VSMCs stimulated by platelet-derived growth factor (PDGF)-BB in the presence or absence of lovastatin.

METHODS

Quiescent VSMCs, with and without lovastatin (20 microM) pretreatment for nine hours, were stimulated by PDGF-BB (25 ng/ml). The incorporation of tritiated thymidine was done to assess DNA synthesis. VSMC lysates were obtained every 6 hours for up to 36 hours after stimulation and were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis using relevant polyclonal antibodies. Autoradiograms were analyzed using a densitometer.

RESULTS

The peak expression of cyclins DPDGF stimulation, respectively. Concomitant expression of CDK4 and CDK2 was also observed. The expression of p27Kip1, by contrast, was reduced in association with DNA synthesis. Lovastatin suppressed DNA synthesis and reduced the expression of cyclin DDK4 and CDK2 expression was unaffected by lovastatin treatment.

CONCLUSIONS

<em>PDGF</em>-BB induces cyclins <em>D</em>1 and E prior to the onset of <em>D</em>NA synthesis in VSMCs. Lovastatin may suppress <em>D</em>NA synthesis in VSMCs by inducing p27Kip1 and reducing expression of cyclins <em>D</em>1 and E.

Objective: To explore the effect of high-voltage electrical burn on platelet function and rheological behavior in rats and the interventive effect of Xuebijing. Methods: A total of 280 Sprague Dawley rats of clean grade (aged 8-10 weeks, male and female unlimited) were divided into sham injury group, simple electrical burn group, electrical burn+ saline group, and electrical burn+ Xuebijing group according to the random number table, with 70 rats in each group. Rats in sham injury group were not conducted with electrical current to cause sham injury. Rats in the other three groups were given electrical current with output voltage of 2 kV and current intensity of (1.92 ± 0.24) A for 3 s, which caused high-voltage electrical burn wounds, each with an area of 1 cm×1 cm distributed in the left forelimb at the current inlet and the right hindlimb at the current outlet respectively. Rats in sham injury group and simple electrical burn group were not treated after injury. At post injury minute 2 and on post injury day (PID) 1, 2, 3, 4, 5, and 6, rats in electrical burn+ saline group and electrical burn+ Xuebijing group were intraperitoneally injected with 6 mL/kg saline and 6 mL/kg Xuebijing, respectively. Survival conditions of rats were recorded during the experiment. At 15 min before injury and at post injury hour (PIH) 1, 8, 24, 48, 72, and on PID 7, 10 rats in each group were respectively selected according to the random number table to sacrifice after collection of 5 mL blood under the direct vision of heart. Blood in the volume of 0.05 mL from each rat was taken to make blood smear, and platelet aggregation number was counted under 400 fold field of view using multiple projection microscope. The remaining blood samples were centrifuged to collect supernatant, and the content of platelet-derived growth factor (PDGF), thrombopoietin (TPO), and platelet activating factor (PAF) was detected by enzyme-linked immunosorbent assay. Data were statistically analyzed with analysis of variance for factorial design and Student-Newman-Keuls method. Results: All rats in sham injury group and simple electrical burn group survived during the experiment. One rat in electrical burn+ saline group died on PID 6, and one rat on PID 5 and one rat on PID 6 died in electrical burn+ Xuebijing group. The levels of all indexes among the 4 groups were close at 15 min before injury. The serum content of PDGF, TPO, and PAF and platelet aggregation number of rats in the three electrical burn groups at all time points after injury were higher or more than those in sham injury group, and the first three indexes reached the peak at PIH 8. The serum platelet aggregation number of rats in simple electrical burn group reached the peak at PIH 48, and that in electrical burn+ saline group and electrical burn+ Xuebijing group reached the peak at PIH 72. Among them, the serum content of PDGF of rats in electrical burn+ Xuebijing group at PIH 48, 72 and on PID 7 ((12.8±4.0), (11.6±4.4), (11.0±3.6) ng/mL, respectively) was close to that in sham injury group ((10.4±2.0), (10.4±2.5), (9.8±3.3) ng/mL, respectively, P>0.05). The serum content of TPO of rats in electrical burn+ Xuebijing group at PIH 24, 72 and on PID 7 ((200±52), (192±36), (193±32) ng/mL, respectively) was close to that in sham injury group ((182±30) , (184±41), (183±33) ng/mL, respectively, P>0.05). The serum content of PDGF, TPO, and PAF and platelet aggregation number of rats in electrical burn+ Xuebijing group at every time point after injury was generally lower or less than that in electrical burn+ saline group and simple electrical burn group. Conclusions: Application of Xuebijing treatment after high-voltage electrical burn can decrease the content of PDGF, TPO, and PAF in the serum and reduce the number of platelet aggregation, thereby inhibit platelet activation and improve platelet rheology.

目的： 探讨高压电烧伤对大鼠血小板功能及流变行为的影响及血必净的干预作用。 方法： 将280只清洁级8～10周龄SD大鼠(雌雄不限)按照随机数字表法分为假伤组、单纯电烧伤组、电烧伤＋盐水组、电烧伤＋血必净组，每组70只。假伤组大鼠不通电致假伤；其余3组大鼠给予输出电压2 kV、电流强度(1.92±0.24)A持续通电3 s，造成电流入口左前肢、电流出口右后肢各1 cm×1 cm大小高压电烧伤创面。假伤组与单纯电烧伤组大鼠伤后不予处理；伤后2 min及1、2、3、4、5、6 d，电烧伤＋盐水组、电烧伤＋血必净组大鼠分别腹腔注射生理盐水6 mL/kg、血必净6 mL/kg。记录实验过程中大鼠的存活情况。伤前15 min及伤后1 h、8 h、24 h、48 h、72 h、7 d，每组按随机数字表法取10只大鼠经心脏直视采血5 mL后处死。取0.05 mL血液制成活血涂片，采用多项投影显微镜于400倍视野下计数血小板聚集数；其余血样离心收集上清液，采用酶联免疫吸附测定法检测血小板源性生长因子(PDGF)、血小板生成素(TPO)、血小板活化因子(PAF)含量。对数据行析因设计方差分析、SNK法检验。 结果： 假伤组和单纯电烧伤组大鼠在实验过程中均存活，电烧伤＋盐水组大鼠于伤后6 d死亡1只，电烧伤＋血必净组大鼠于伤后5、6 d各死亡1只。各组大鼠伤前15 min各指标水平相近。3组电烧伤大鼠伤后各时间点血清PDGF、TPO、PAF含量及血小板聚集数均高于或多于假伤组，前3个指标于伤后8 h达峰值；单纯电烧伤组大鼠血清血小板聚集数于伤后48 h达峰值，电烧伤＋盐水组和电烧伤＋血必净组大鼠血清血小板聚集数于伤后72 h达峰值。其中电烧伤＋血必净组大鼠血清PDGF含量于伤后48、72 h及7 d[分别为(12.8±4.0)、(11.6±4.4)、(11.0±3.6)ng/mL]与假伤组[分别为(10.4±2.0)、(10.4±2.5)、(9.8±3.3)ng/mL]接近(P>0.05)，血清TPO含量于伤后24、72 h及7 d[分别为(200±52)、(192±36)、(193±32)ng/mL]与假伤组[分别为(182±30)、(184±41)、(183±33)ng/mL]接近(P>0.05)。电烧伤＋血必净组大鼠伤后各时间点血清PDGF、TPO、PAF含量及血小板聚集数普遍低于或少于电烧伤＋盐水组和单纯电烧伤组。 结论： 大鼠高压电烧伤后应用血必净治疗可降低血清中PDGF、TPO、PAF含量及减少血小板聚集数，抑制血小板活化，从而改善血小板流变性。.

Keywords: Burns, electric; Platelet activating factor; Platelet-derived growth factor; Thrombopoietin; Xuebijing.

Objective: To study the role of human antigen R (HuR) regulated transforming growth factor β1 (TGF-β1) expression in airway smooth muscle cells under the stimulation of platelet-derived growth factor (PDGF). Methods: Airway smooth muscle (ASM) cells were cultured at 37 ℃ and 5% CO2 in dulbecco's modified eagle medium (DMEM) cell medium. Cells at passages between 4 and 11 were divided into different groups according to the different compounds added. For control group, no compounds were administrated. For PDGF group, cells were stimulated with PDGF (20 μg/L) and cultured for an additional time. Cells were harvested and real-time PCR was used to measure mRNA level and Western blotting to detect protein level of HuR and TGF-β1 in ASM cells for different groups. Cells were divided into HuR siRNA group and control group. RNA-interference was used to determine whether lowering HuR expression could decrease PDGF-induced TGF-β1 expression in HuR siRNA group and control group after the stimulation of PDGF for indicated times. Western blotting analysis was used to test the expression of TGF-β1 after interrupting HuR expression. The concentration of TGF-β1 in the cultured serum of HuR siRNA group and control group for 0, 6, 12 h under the stimulation of PDGF was measured by enzyme-link immunosorbent assay (ELISA). Cells were divided into control group, control+ PDGF 6 h group, HuR siRNA group and HuR siRNA+ PDGF 6 h group, then the half-life of TGF-β1 mRNA in different groups was determined by treating ASM cells with the transcriptional inhibitor actinomycin D (10 mg/L) for 0, 4, 8 and 12 h. Results: PDGF treatment for 0, 6, 12 and 24 h significantly promoted HuR mRNA and protein expression and the relative levels were 1.00±0.00, 1.35±0.14, 1.73±0.17, 2.07±0.10; 0.51±0.10, 0.67±0.05, 0.83±0.07, 0.95±0.02 (all P<0.05). Similar alterations could also be demonstrated at TGF-β1 mRNA and protein. The relative expression was 1.00±0.00, 1.27±0.06, 1.60±0.10, 1.87±0.10; 0.72±0.09, 0.87±0.07, 1.13±0.12, 1.33±0.05 (6 h versus 12 h and 12 h versus 24 h, P<0.05). HuR expression decreased 21.9% in HuR siRNA group compared with control group under the stimulation of PDGF for 12 h. HuR silencing also decreased PDGF-induced TGF-β1 over-expression in ASM cells. In the control group, the relative protein levels of PDGF treatment for 0, 6 and 12 h were 0.70±0.05, 0.89±0.06, 1.06±0.05 and the protein levels in HuR siRNA group were 0.67±0.09, 0.77±0.03, 0.89±0.05 (all P<0.05). The concentration of TGF-β1 in the cultured serum was measured by ELISA and the outcomes were (773.33±16.32, 877.97±16.03, 3 060.34±82.53) ng/L in control group for 0, 6 and 12 h. Under the same condition, the outcomes of HuR siRNA group were (277.33±9.93, 407.77±7.14, 828.05±11.67) ng/L (both P<0.05). Actinomycin D disturbed the process of transcription and the half-life of TGF-β1 mRNA in HuR siRNA group showed no significant change compared with the HuR siRNA+ PDGF 6 h group (P>0.05). However, compared with control group and control + PDGF 6 h group, the half-life of TGF-β1 mRNA in HuR siRNA group showed significant change (P<0.05). Conclusions: PDGF can elevate TGF-β1 expression in ASM cells. HuR regulates TGF-β1 expression by promoting its mRNA stability.

Platelet-derived growth factor is commonly known as a mitogen. Many research data suggest a role for PDGF-beta R in the mitogenic response of mesangial cells. There are four members of PDGF family known as PDGF-A chain, PDGF-B chain, PDGF-C chain and PDGF-D chain, which in active forms are dimers. As far as two receptors PDGF-alpha R and PDGF-beta R are known to bind PDGF. There is a difference in binding affinity of various forms of PDGF by these receptors. Two different promotors P1 and P2 can be used for PDGF-alpha R gene transcription. There are several different haplotypes of promotor P1 sequence. Transcription of PDGF-alpha R gene is under control of many factors. Interaction between a receptor and its ligand includes receptor dimerisation and autophosphorylation of tyrosine residues. PDGF AA is unique in that it can only be bound by alpha-receptor dimer. PDGF-AA expression has been confirmed in the normal kidney, as well as in several renal diseases. Although the expression of PDGF-alpha R has been found to accompany that of PDGF-AA, its actual relevance for the development of the glomerular pathology is not clear.

OBJECTIVE

To study the effect of angiotensin-(1-7) on the kidney of diabetic rats by observing the mRNA expression of PDGF and TGF-beta1.

METHODS

SD rats were divided into three groups: Group C (uni-nephrectomy control group), Group D (diabetic model control group), Group T (Ang-(1-7) treated group). We evaluated blood glucose,urea nitrogen, creatinine and urine albumin excretion respectively, studied the renal morphology by light microscope, and detected the gene expression of PDGF, TGF-beta1 in renal tissue by RT-PCR technique.

RESULTS

There was significant difference between the group D and T about the RW/BW, renal morphology, the total urine protein and the mRNA expression of PDGF and TGF-beta1.

CONCLUSIONS

Ang-(1-7) can relieve the renal process of diabetic rats.

Platelet-derived growth factor (PDGF) is a potent mesenchymal cell mitogen and chemoattractant involved in the pathogenesis of fibroproliferative diseases. There are four known PDGF ligand isoforms designated A-D, two of which, C and D, were only recently discovered. We have identified a splicing variant in the PDGF-D isoform that occurs in mice, but not in humans. The presence of the splicing variant in murine PDGF-D appears to be due to an aberration in the splicing site at the junction of exons 5 and 6. The splicing variant results in a deletion predicted to have significant effects on peptide activity since it results in the deletion of bases within the cysteine knot domain that are important for peptide dimerization and receptor binding. It is important to appreciate differences between murine and human PDGF gene expression because PDGF is a key mitogen in the pathogenesis of fibrosis and mice are commonly employed as models for human disease.

Following the publication of the above paper, a concerned reader drew to the Editor's attention that several figures contained data that bore striking similarities to data published in other papers; notably, the western blot data shown in Fig. 6 appeared to have been presented in other studies, notably in Fig. 7B of another paper published around the same time and written by different authors based at different research institutions [Li P, Zhang Z, Zhang F, Zhou H and Sun W: Effects of 3‑tetrazolyl methyl‑3‑hydroxy‑oxindole hybrid (THOH) on cell proliferation, apoptosis, and G2/M cell cycle arrest occurs by targeting platelet‑derived growth factor D (PDGF‑D) and the MEK/ERK signaling pathway in human lung cell lines SK‑LU‑1, A549, and A‑427. Med Sci Monit 24: 4547‑4554, 2018]. Furthermore, cellular images featured in Fig. 2A and B of the above paper appeared in Fig. 2 of the following paper, albeit the data were presented in a different field of view: Yu L, Zhou G‑Q and Li D‑C: MiR‑136 triggers apoptosis in human gastric cancer cells by targeting AEG‑1 and BCL2. Eur Rev Med Pharmacol Sci 22: 7251‑7256, 2018. After having conducted an independent investigation in the Editorial Office, the Editor of International Journal of Molecular Medicine has determined that this article should be retracted from the Journal on account of a lack of confidence concerning the originality and the authenticity of the data. The authors were asked for an explanation to account for these concerns, but the Editorial Office never received any reply. The Editor regrets any inconvenience that has been caused to the readership of the Journal. [the original article was published on International Journal of Molecular Medicine 41, 3485-3492, 2018; DOI: 10.3892/ijmm.2018.3531].

Long-term bone defects are a key clinical problem. Autogenous bone graft remains the gold standard for the treatment of these defects; however, improving the osteogenic properties and reducing the amount of autogenous bone is challenging. Autologous platelet-rich plasma (PRP) has been widely considered for treatment, due to its potentially beneficial effect on bone regeneration and vascularization. The aim of the present study was to explore the effects of autogenous bone particles combined with PRP on repairing segmental bone defects in rabbits. Briefly, a critical-size diaphyseal radius defect was established in 45 New Zealand White rabbits. Animals were randomly divided into four groups, according to the different implants: Group A, empty bone defect; group B, PRP; group C, autogenous bone particles + bone mesenchymal stem cells (BMSCs) on the left radius; group D, autogenous bone particles + PRP + BMSCs on the right radius. Bone samples were collected and further analyzed using X-ray, histology and histomorphometry 4, 8 and 12 weeks post-surgery. In addition, the effect of PRP on cell proliferation was detected by Cell Counting Kit-8 and the concentrations of growth factors (GFs), transforming GF (TGF)-β1 and platelet-derived GF (PDGF), in PRP were verified by ELISA. X-ray, histology and histomorphometry data revealed that the fraction area of the newly formed bone was larger in group D. In addition, PRP could improve cell proliferation, osteogenic differentiation and the release of GFs, TGF-β1 and PDGF-AB. In conclusion, these findings indicated that an autogenous bone particle + PRP + BMSC scaffold may be used as a potential treatment strategy for segmental defects in humans.

Keywords: BMSCs; PRP; autogenous bone particles; bone defect; bone regeneration.

When the receptors for platelet-derived growth factor (PDGF) are activated they aggregate, become tyrosine-phosphorylated and elicit a cascade of down-stream signals, including mobilization of Ca2+ from intra- and extracellular stores. Receptor mobility in the plane of the membrane is a prerequisite for receptor aggregation and further signalling. Using human foreskin fibroblasts (AG 1523) and fluorescence recovery after photobleaching (FRAP), we therefore assessed the lateral mobility characteristics of PDGF-beta2 receptors by their diffusion coefficient (D), and fraction of mobile receptors (R). This was done on cells stimulated with either normal human serum (NHS) or PDGF under different Ca2+-conditions. The results suggest that both intra- and extracellular free Ca2+ influence the mobility characteristics of the PDGF-beta2 receptor. Interestingly, the extracellular Ca2+ seems to impose general restrictions on the mobility of receptors, since R increased when extracellular Ca2+ was quenched with EGTA, whereas intracellular clamping of Ca2+ transients with MABTAM (BAPT/AM) primarily affected D. When both intra- and extracellular Ca2+ were quenced, D remained low and R high, further supporting the proposition that they achieve distinct effects. Inhibition of tyrosine phosphorylation with Erbstatin, partly inhibited the NHS effects and released PDGF-induced receptor immobilization. Ratio imaging with Fura-2 displayed that both NHS and PDGF induced changes in intracellular free [Ca2+]. In view of the present data it might have important effects on the state of the receptor in the membrane, for instance by regulating its lateral mobility, communication with other receptors and signalling functions in the membrane.

OBJECTIVE

Therapeutic exposure to high doses of radiation can severely impair organ function due to ablation of stem cells. Normal tissue injury is a dose-limiting toxicity for radiation therapy (RT). Although advances in the delivery of high precision conformal RT has increased normal tissue sparing, mitigating and therapeutic strategies that could alleviate early and chronic radiation effects are urgently needed in order to deliver curative doses of RT, especially in abdominal, pelvic and thoracic malignancies. Radiation-induced gastrointestinal injury is also a major cause of lethality from accidental or intentional exposure to whole body irradiation in the case of nuclear accidents or terrorism. This review examines the therapeutic options for mitigation of non-hematopoietic radiation injuries.

RESULTS

We have developed stem cell based therapies for the mitigation of acute radiation syndrome (ARS) and radiation-induced gastrointestinal syndrome (RIGS). This is a promising option because of the robustness of standardized isolation and transplantation of stromal cells protocols, and their ability to support and replace radiation-damaged stem cells and stem cell niche. Stromal progenitor cells (SPC) represent a unique multipotent and heterogeneous cell population with regenerative, immunosuppressive, anti-inflammatory, and wound healing properties. SPC are also known to secrete various key cytokines and growth factors such as platelet derived growth factors (PDGF), keratinocyte growth factor (KGF), R-spondins (Rspo), and may consequently exert their regenerative effects via paracrine function. Additionally, secretory vesicles such as exosomes or microparticles can potentially be a cell-free alternative replacing the cell transplant in some cases.

CONCLUSIONS

This review highlights the beneficial effects of SPC on tissue regeneration with their ability to (a) target the irradiated tissues, (b) recruit host stromal cells, (c) regenerate endothelium and epithelium, (d) and secrete regenerative and immunomodulatory paracrine signals to control inflammation, ulceration, wound healing and fibrosis.

BACKGROUND

15-Deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2), one of the major metabolites from prostaglandin DD,L-lactide-co-glycolide) nanocapsules (15d-PGJ2-NC) on inflammatory responses and bone regeneration in local bone defect.

METHODS

The study was conducted on 96 Wistar rats from June 2014 to March 2016. Saline, unloaded nanoparticles, free 15d-PGJ2or 15d-PGJ2-NC, were delivered through a collagen vehicle inside surgically created transcortical defects in rat femurs. Interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) levels in the surrounding soft tissue were analyzed by Western blot and in the defect by quantitative real-time polymerase chain reaction over 14 days. Simultaneously, bone morphogenetic protein-6 (BMP-6) and platelet-derived growth factor-B (PDGF-B) messenger RNA (mRNA) in the defect were examined. New bone formation and EphrinB2 and osteoprotegerin (OPG) protein expression in the cortical defect were observed by Masson's Trichrome staining and immunohistochemistry over 28 days. Data were analyzed by one-way analysis of variance. Least-significant difference and Dunnett's T3 methods were used with a bilateral P< 0.05.

RESULTS

Application of l5d-PGJ2-NC (100 μg/ml) in the local bone defect significantly decreased IL-6, IL-1β, and TNF-α mRNA and protein, compared with saline-treated controls (P < 0.05). l5d-PGJ2-NC upregulated BMP-6 and PDGF-B mRNA (P < 0.05). New bone formation was observed in the cortical defect in l5d-PGJ2-NC-treated animals from 7th day onward (P < 0.001). Expression of EphrinB2 and OPG presented early on day 3 and persisted through day 28 in 15d-PGJ2-NC group (P < 0.05).

CONCLUSIONS

Stable l5d-PGJ2-NC complexes were prepared that could attenuate IL-6, IL-1β, and TNF-α expression, while increasing new bone formation and growth factors related to bone regeneration.

Although advances in the prediction and management of ovarian hyperstimulation syndrome (OHSS) have been introduced, complete prevention is not yet possible. Previously, we and other authors have shown that vascular endothelial growth factor, angiopoietins (ANGPTs) and sphingosine-1-phosphate are involved in OHSS etiology. In addition, we have demonstrated that ovarian protein levels of platelet-derived growth factor (PDGF) ligands -B and -D decrease in an OHSS rat model, whilst PDGFR-β and ANGPT2 remain unchanged. In the present work, we investigated the role of PDGF-B in OHSS by evaluating ligand protein levels in follicular fluid (FF) from women at risk of developing OHSS and by using an immature rat model of OHSS. We demonstrated that PDGF-B and PDGF-D are lower in FF from women at risk of developing OHSS compared to control patients (p < 0.05). In the OHSS rat model, PDGF-B (0.5 µg/ovary) administration decreased ovarian weight (p < 0.05), reduced serum progesterone (p < 0.05) and lowered the percentage of cysts (p < 0.05), compared to untreated OHSS rats, but had no effect on the proportion of follicles or corpora lutea (CL). PDGF-B treatment also restored the expression of steroidogenic acute regulatory protein (p < 0.05) and P450 cholesterol side-chain cleavage enzyme (p < 0.01) to control levels. In addition, PDGF-B increased the peri-endothelial cell area in CL and cystic structures, and reduced vascular permeability compared to untreated OHSS ovaries. Lastly, PDGF-B increased the levels of junction proteins claudin-5 (p < 0.05), occludin (p < 0.05) and β-catenin (p < 0.05), while boosting the extracellular deposition of collagen IV surrounding the ovarian vasculature (p < 0.01), compared to OHSS alone. In conclusion, our findings indicate that PDGF-B could be another crucial mediator in the onset and development of OHSS, which may lead to the development of novel prediction markers and therapeutic strategies.

Keywords: angiogenesis; infertility; ovarian hyperstimulation syndrome; ovary; ovulation induction; platelet-derived growth factor B; vascular leakage.

Many years have passed since domestic water fluoridation was adopted to reduce the incidence of caries in developed countries; however, since there is an additional dose of fluorides ingested with foods and drinks prepared with such waters, the problem has emerged of possible adverse effects on health associated to them, so that in some countries fluorine integrator selling is allowed only with preventive medical prescription. Owing to the affinity for calcifited tissues, fluorine has a powerful effect on bone cellular order (mediated by growth factors' upregulation system IGF-2, TGF-beta, PDGF, bFGF, EGF, BMP-2 and PTH), on function and length, since it can provoke chronic joints-pain, ligaments-calcification, osteosclerosis. Moreover, sodium-fluoride may cause adverse effects on testicular activity (connected to oxidative-stress depending on increased activity of peroxidases and catalases) due to inhibition of 2 androgenesis-regulator enzymes DELTA(5)b-HSD and 17beta-HSD. Furthermore, insoluble gut formed calcium-fluoride may be responsible for hypocalcemia inducing a secondary hyperparathyroidism with bone matrix resorption, osteoporosis, osteomalacia and, perhaps, lowered level of phosphorus. At encephalic level, then, high doses of fluorine cause the onset of neurological symptoms and of a decreased spontaneous motor activity due to a reduction in the number of nicotinic acetylcholine receptors. Nevertheless, epidemiological studies about fluoride toxicity have established that such oligoelement may be safely used at odontoiatric dosages.

Platelet-derived growth factors (PDGFs) are key regulators of mesenchymal cells in vertebrate development. To what extent PDGFs also exert beneficial homeostatic or reparative roles in adult organs, as opposed to adverse fibrogenic responses in pathology, are unclear. PDGF signaling plays critical roles during heart development, during which forced overexpression of PDGFs induces detrimental cardiac fibrosis; other studies have implicated PDGF signaling in post-infarct myocardial repair. Different PDGFs may exert different effects mediated through the two PDGF receptors (PDGFRα and PDGFRβ) in different cell types. Here, we assessed responses induced by five known PDGF isoforms in the adult mouse heart in the context of adenovirus vector-mediated inflammation. Our results show that different PDGFs have different, in some cases even opposing, effects. Strikingly, whereas the major PDGFRα agonists (PDGF-A and -C) decreased the amount of scar tissue and increased the numbers of PDGFRα-positive fibroblasts, PDGFRβ agonists either induced large scars with extensive inflammation (PDGF-B) or dampened the adenovirus-induced inflammation and produced a small and dense scar (PDGF-D). These results provide evidence for PDGF isoform-specific inflammation-modulating functions that may have therapeutic implications. They also illustrate a surprising complexity in the PDGF-mediated pathophysiological responses.

The new knowledge in molecular biology and pathophysiology of chronic myeloid leukemia enabled the development of imatinib mesylate (Glivec, formerly STI571). Imatinib potently inhibits several protein tyrosine kinases, including BCR-ABL, c-Kit, and PDGF receptor. Imatinib blocks the phosphorylation of downstream target proteins and interrupts the malignant transformation leading to the development of CML. Phase I and II studies demonstrated that imatinib is highly effective and well tolerated in all phase of CML. We got our experience with imatinib on more than two-year monitoring 34 patients within the Expanded Access Study CST1571 0113. Imatinib 400 mg/d was administered orally to 10 women and 24 men in median age of 53 years (22-70) who were hematologically (n = 9) or cytogenetically (n = 13) resistant, cytogenetically refractory (n = 3) or intolerant (n = 9) to interferon alpha. The median follow-up time was 97.5 weeks (23-115), the median time from CML diagnosis to the start of the study was 32.3 months (6-140.5). Complete hematologic response was achieved in 33 of 34 (97%) pts, total major cytogenetic response (complete plus major) in 21 of 33 (63%) pts. Cytogenetic relapse was observed in 2 of 33 pts (6%), cytogenetic progression in 4 (12%) pts. Non-hematologic toxicity was mild (grade 1 or 2) and no patient was excluded from the study due to it. Hematological toxicity grade 3 limited dose of imatinib in 26% of patients and probably caused lower rate of cytogenetic responses in heavy pretreated patients. Both quantitative RT-PCR methods (competitive RT-PCR and real-time RT-PCR Light-Cycler) were found useful to monitor patients with CML on imatinib therapy. Our results confirmed high efficacy and safety of imatinib in late-chronic phase CML patients failing prior interferon therapy. The lower incidence of hematological toxicity and higher rate of cytogenetic responses in patients treated with imatinib in early-chronic phase CML justify according to our opinion the recommendation to administer imatinib early after the diagnosis of CML in patients who are not indicated for allogeneic transplantation.