MOTS-c accelerates bone fracture healing by stimulating osteogenesis of bone marrow mesenchymal stem cells via positively regulating FOXF1 to activate the TGF-ß pathway.

The article "MOTS-c accelerates bone fracture healing by stimulating osteogenesis of bone marrow mesenchymal stem cells via positively regulating FOXF1 to activate the TGF-ß pathway, by F.-B. Weng, L.-F. Zhu, J.-X. Zhou, Y. Shan, Z.-G. Tian, L.-W. Yang, published in Eur Rev Med Pharmacol Sci 2019; 23 (24): 10623-10630-DOI: 10.26355/eurrev_201912_19759-PMID: 31858528" has been withdrawn from the authors due to inaccuracies during the process of organizing the images. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19759.

[CT manifestations and prognosis of acute paraquat induced lung injury].

Objective: To investigate the CT features of lung injury induced by paraquat poisoning and its relationship with prognosis, and to provide reference for the judgment of the condition and prognosis of paraquat poisoning. Methods: 146 cases of paraquat poisoning patients were treated in the Third People's Hospital of Xuzhou City from January 2013 to April 2016. The cases were divided into mild group, moderate-severe group and fulminant group according to the concentration of paraquat in urine. The clinical data and CT imaging findings were analyzed and reconstructed in three-dimensional reconstruction. The extent of the lesion was observed and the relationship between CT and prognosis was explored. Results: Paraquat lung injury has many manifestations on CT images, and it's performance can be intersecting at the same time. Early lesions lighter cases, late CT imaging lesions can be completely absorbed or residual fibrosis, the prognosis was good; the early lesion was pulmonary consolidation, pleural effusion cases, the late CT image was usually pleural thickening and bronchiectasis, the prognosis was relatively good; early lesions were large patches of ground glass opacity cases, finally, pulmonary fibrosis was common, the mortality rate of 56.57%. There were significant differences in the extent of lung injury between different groups (P<0.001) , and the difference in mortality was statistically significant when the lung injury was different (P<0.001) . Multivariate stepwise Logistic regression analysis showed that ground-glass opacity (OR value=2.013) , interstitial lung fibrosis (OR=3.779) and mediastinal emphysema (OR=33.118) were risk factors for death of lung injury caused by paraquat poisoning (P<0.05) . Conclusion: There were many manifestations on CT images of paraquat lung injury, and the manifestations of paraquat lung injury can be intersecting at the same time. The pulmonary manifestations and outcomes of different paraquat types were different. The CT manifestations of lung injury in paraquat poisoning were mainly exudative changes at early stage, and can be gradually absorbed or evolved into interstitial changes at later stage. The cumulative damage range can be used as a reference for evaluating the prognosis. Ground-glass opacity, interstitial pulmonary fibrosis and mediastinal emphysema are the risk factors for death of lung injury caused by paraquat poisoning.

MOTS-c accelerates bone fracture healing by stimulating osteogenesis of bone marrow mesenchymal stem cells via positively regulating FOXF1 to activate the TGF-ß pathway.

OBJECTIVE: To elucidate the function of MOTS-c in accelerating bone fracture healing by inducing BMSCs differentiation into osteoblasts, as well as its potential mechanism. MATERIALS AND METHODS: Primary BMSCs were extracted from rats and induced for osteogenesis. The highest dose of MOTS-c that did not affect BMSCs proliferation was determined by CCK-8 assay. After 7-day osteogenesis, the relative levels of ALP, Bglap, and Runx2 in MOTS-c-treated BMSCs influenced by FOXF1 were examined. ALP staining and alizarin red S staining in BMSCs were performed as well. The interaction between FOXF1 and TGF-ß was analyzed by ChIP assay. At last, rescue experiments were performed to uncover the role of FOXF1/TGF-ß axis in MOTS-c-induced osteogenesis. RESULTS: 1 µM MOTS-c was the highest dose that did not affect BMSCs proliferation. MOTS-c treatment upregulated the relative levels of ALP, Bglap, and Runx2, and stimulated mineralization ability in BMSCs, which were attenuated by the silence of FOXF1. TGF-ß was proved to interact with FOXF1, and its level was positively mediated by FOXF1. The silence of FOXF1 attenuated the accelerated osteogenesis and TGF-ß upregulation in BMSCs because of MOTS-c induction, and these trends were further reversed by the overexpression of TGF-ß. CONCLUSIONS: MOTS-c treatment markedly induces osteogenesis in BMSCs. During MOTS-c-induced osteogenic progression, the upregulated FOXF1 triggers the activation of TGF-ß pathway, thus accelerating bone fracture healing.

MicroRNA-615-3p promotes the osteoarthritis progression by inhibiting chondrogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE: To investigate whether microRNA-615-3p participates in the development and progression of osteoarthritis by regulating chondrogenic differentiation of bone marrow mesenchymal stem cells. MATERIALS AND METHODS: Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat bone marrow and identified by flow cytometry. After chondrogenic differentiation was induced in BMSCs, expression levels of chondrogenic-specific genes were then detected by quantitate Real-time polymerase chain reaction (qRT-PCR). Expression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Protein expression of SOX9 after overexpression or knockdown of microRNA-615-3p was detected by Western blot, respectively. RESULTS: MicroRNA-615-3p was down-regulated in the process of chondrogenic differentiation of BMSCs. The mRNA expressions of chondrogenic-specific markers, COL2A1, COL10A1, ACAN and MATN3 were decreased after microRNA-615-3p overexpression in BMSCs. Overexpressed microRNA-615-3p down-regulated protein expression of SOX9. Expression levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-α (IL-α) were increased after overexpression of microRNA-615-3p, while inhibition of microRNA-615-3p expression obtained the opposite result. In addition, overexpression of SOX9 rescued the effect induced by microRNA-615-3p on inflammatory cytokines. CONCLUSIONS: MicroRNA-615-3p participates in the development and progression of osteoarthritis by increasing the expressions of inflammatory cytokines and inhibiting chondrogenic differentiation of BMSCs.

MicroRNA-337-5p participates in the development and progression of osteosarcoma via ERBB, MAPK and VEGF pathways.

OBJECTIVE: To investigate the role of microRNA-337-5p in osteosarcoma (OS) and its underlying mechanism. PATIENTS AND METHODS: The microRNA (microRNA-337-5p) that may be related to OS development was screened out by GEO (Gene Expression Omnibus) database. Survival analysis and ROC curve were performed according to microRNA-337-5p expressions in OA patients. Besides, the correlation between microRNA-337-5p expression and clinical parameters was evaluated by Chi-square analysis. Cox regression analysis was performed to detect the relationship between the overall survival and clinical parameters of OA patients. Subsequently, enriched functions and pathways of microRNA-337-5p were predicted by GESA (gene enrichment sets analysis). MicroRNA-337-5p expression was detected in 65 OS tissue samples and 30 normal tissue samples by qRT-PCR (quantitative Real-Time Polymerase Chain Reaction). For in vitro experiments, after microRNA-337-5p mimics or microRNA-337-5p inhibitor was transfected into OS cells, proliferative and invasive abilities were detected by CCK-8 (Cell Counting Kit-8) and transwell assay, respectively. Finally, Western blot was used to explore the underlying mechanism of microRNA-337-5p in regulating OS. RESULTS: MicroRNA-337-5p was overexpressed in serum and tissue samples of OS patients, which was valuable in diagnosing OS. Besides, microRNA-337-5p expression was correlated with the overall survival and necrosis range of OA patients, whereas not correlated with age and sex. GESA indicated that microRNA-337-5p was enriched in ERBB, MAPK, and VEGF pathways. In vitro experiments indicated elevated proliferative and invasive abilities in MG63 and U2OS cells after microRNA-337-5p overexpression. Furthermore, increased expressions of ERBB2, Erk1/2, and VEGF121 were observed in OS cells after microRNA-337-5p overexpression. CONCLUSIONS: MicroRNA-337-5p is upregulated in OS tissues, which is an independent prognostic factor in OS. Overexpressed microRNA-337-5p can promote proliferative and invasive abilities of OS cells via activating ERBB, MAPK, and VEGF pathways.

[Epidemiological characteristics of pesticide poisoning in Xuzhou city from 2005 to 2017].

Objective: To understand the characteristics and causes of pesticide poisoning in Xuzhou city, and provide basis for formulating prevention and control measures. Methods: The cases of pesticide poisoning in Xuzhou City from 2005 to 2017 were collected from "Pesticide Poisoning Report Card" . The data were analyzed and assessed by EpiData. The SPSS 22.0 software was used for statistical analysis. Results: During the thirteen years, there were a total of 8092 cases of pesticide poisoning, among which, the number of occupational pesticide poisoning was 1 408, accounting for 17.4% of the total number of cases, 14 patients died, the case fatality rate was 0.1%. There were 2, 992 cases of male poisoning, accounting for 36.97% of the total number of cases, and 5, 100 cases of female poisoning, accounting for 63.03%. There were 6684 non-productive pesticide poisonings, accounting for 82.6% of the total number of cases; 387 deaths occurred, and the mortality rate was 5.8%. Among non-productive poisonings, the incidence of oral pesticide poisoning was 84.3%, and the incidence of accidental poisoning by pesticides was 15.7%. Organophosphorus pesticides poisoning cases accounted for the majority of oral pesticide poisoning cases. The overall incidence of pesticide poisoning showed a downward trend. The age of non-productive pesticide poisoning cases was mainly 15-44 years old, and the number of cases of poisoning were 4 029 cases (60.28%) . With the increase of age, the mortality rate of poisoning cases was higher, especially for those over 60 years old who died of oral pesticide poisoning (40.1%) . The peak of pesticide poisoning began to increase in the second quarter and reached its peak in the third quarter. Conclusion: Although the cases of pesticide poisoning reported in Xuzhou City have been declining in recent years, the situation is still severe. The proportion of oral pesticide suicide accounts for a large proportion, and the mortality rate of elderly and female is relatively high, and the government should pay more attention. Workers should conduct safety education and psychological counseling to improve the knowledge and consciousness of safe use of pesticides.

[Clinical application study of non-invasive ventilation in the treatment of the pneumoconiosis patients with chronic type â ¡ respiratory failure].

Objective: To observe the effect of early intervention and intermittent application of bi level positive air-way pressure ventilation (BiPAP) in patients with pneumoconiosis combined with chronic respiratory failure. Methods: Will meet the diagnostic criteria of pneumoconiosis in GBZ70-2009< >, the blood gas analysis in patients with chronic type II re-spiratory failure in 62 cases were randomly divided into rehabilitation treatment group 32 cases, control group of 30 cases. Pa-tients in the observation group were treated by on-invasive ventilation, while the control group were treated by the convention-al treatment. The data such as arterial blood gas、pulmonary function、the grade about dyspnea and echocardiography was col-lected from the both group before the beginning of the treatment and after the three months. Results: the PaO2 level、FEV1.0、FEV1.0%、the grade of dyspnea and the Right Ventricular Ejection Fractions were not significantly different between the experi-mental group and the control before the start of the treatment (P>0.05) . After the three month treatment, the PaO2 level of the observation was significantly lower the control (P<0.05) . The data about FEV1.0、FEV1.0% and the Right Ventricular Ejection Fractions were higher than the control group (P<0.05) . Conclusion: Non-invasive ventilation has exactly effect in the treat-ment of the pneumoconiosis patients with Chronic Respiratory Failure. It can improve the function of the heart and lung and ease the pain of patients.

Cell-based therapies for experimental diabetic nephropathy: a systematic review and meta-analysis.

Study has shown that stem cell–based therapies are promising strategies in the treatment of several chronic diseases, but their overall benefit in the treatment of diabetic nephropathy (DN) remains controversial. The purpose of this study is to summarize the evidence of the effect of cell-based therapy in the treatment of DN to guide future clinical trials. We searched PubMed, EmBase, and the Cochrane Library for studies from the inception of cell-based therapies up to July 2015. We included animal trials that reported the effects of cell-based therapy on kidney function, cardiovascular risk factors, and body factors. A random-effects model was used to process the data, and the standard mean difference (SMD) was used to evaluate the efficacy of cell-based therapy. We included eight studies that reported data on 159 mice. Overall, we noted that cell-based therapies were associated with significantly reduced plasma creatinine level (P = 0.003), glomerular filtration rate (P less than 0.001), plasma glucose level (P = 0.004), serum cholesterol level (P = 0.010), serum triglyceride level (P = 0.032), plasma urea level (P less than 0.001), proteinuria (P = 0.008), and Cl- fractional excretion (P = 0.023). Furthermore, cell-based therapies were associated with lower kidney weight (P = 0.003), and kidney/body weight (P = 0.004). A sensitivity analysis suggested that cell-based therapy might play an important role in increased body weight. In conclusion, cell-based therapies significantly improve kidney function, cardiovascular risk factors, and body factors in the treatment of DN.

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100 percent; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6 percent, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83 percent, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7 percent ethanol (62 vs 62 percent, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62 percent, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development.

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development.

Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca(2)+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca(2)+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 microg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca(2)+]i oscillations, completion of meiosis and parthenogenetic development.

Effect of conditioned medium of mesenchymal stem cells on the in vitro maturation and subsequent development of mouse oocyte.

Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.

Effect of conditioned medium of mesenchymal stem cells on the in vitro maturation and subsequent development of mouse oocyte

Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.

[Purification and biological activity of rh-leptin expressed in Escherichia coli].

The human leptin was successfully expressed with high level in E. coli under the control of PL promotor. The yield of recombinant protein was over 40% of total cellular protein and expressed as inclusion bodies. The recombinant human leptin (rh-leptin) was purified with gel filtration, anion-exchange and reverse chromatography. Refolding was achieved by gradually reducing denaturant using a diafiltration method. The refolded rh-leptin was characterized by SDS-PAGE, Western-blotting and its first 15 amino acid residues sequence of the N-terminal. The purified product was found to be biologically active, reducing the food intake and body weight gain upon testing in BALB/c mice.

[Significance of the unbalanced expression of Th1/Th2 type cytokines in human glioma].

OBJECTIVE: To study the significance of the unbalanced expression of Th1/Th2 type cytokines in human glioma. METHODS: The gene expressions of Th1/Th2 type cytokines in 62 specimens of human glioma tissues, 4 glioma cell lines, peripheral blood mononuclear cell (PBMC) of 15 glioma patients, 5 specimens of normal adult brain tissue and 5 brain meningioma tissues were detected by semiquantitative reverse transcription polymerase chain reaction. IFN-gamma and IL-2 represent Th1 type cytokines. IL-4, IL-6, IL-10 and IL-13 represent Th2 type cytokines. RESULTS: There were obviously predominant expression of Th2 type cytokines in glioma cell lines (P < 0.01) and specimens of human glioma tissues (P < 0.01). The tendency of distinct expression of Th2 type cytokines in PBMC was also existent. There wasn't obvious discrepancy of the expression of two type cytokines in normal adult brain tissues and meningioma tissues. CONCLUSIONS: It is likely that the switching of Th1/Th2 type cytokines in gliomas as predominant expression of Th2 type cytokine genes is related to the origination of gliomas and the evasion of glioma cells from immune surveillance.

[Influence of adjustment of balance of Th1/Th2 type cytokines on proliferation of glioma cells].

OBJECTIVE: To study the influence of adjustment of balance of Th1/Th2 by external cytokines on proliferation of glioma cells. METHODS: The gene expressions of Th1/Th2 type cytokines in C6, 9L, U251 and SHG44 glioma cells were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). After the cells were induced with IFN-gamma + IL-4 McAb and IL-4 + IFN-gamma McAb respectively, we isolated the total RNA to proceed RT-PCR again. The evaluation of cell proliferation was proceeded by MTT assay method. RESULTS: There was obviously predominant expression of Th2 type cytokines in glioma cell lines (P < 0.01). The expression intensity of IFN-gamma was improved in IFN-gamma + IL-4 McAb groups and Th2 type cytokines were enhanced in IL-4 + IFN-gamma McAb groups. IFN-gamma and IL-4 McAb could cause the switch from Th2 to Th1, and could remarkably inhibit the proliferation of glioma cells in a dose-dependent way (P < 0.01). On the other hand, IL-4 and IFN-gamma McAb could strengthen the switch of Th2, and might stimulate the glioma cell growth, also in a dose-dependent way (P < 0.01). CONCLUSIONS: There is a Th2 preponderance in glioma cells. IFN-gamma and IL-4 McAb could regulate the switch from Th2 to Th0 or Th1, and inhibit the proliferation of glioma cells.

[Differentiation of natural killer cells into two functional subsets: NKh1 and NKh2].

OBJECTIVE: To verify the presence of functional subsets of natural killer cells based on the cytokine production. METHODS: NK cells were purified and cultured in complete RPMI1640 medium in the presence of either IFN gamma + anti-IL-4(classical Th1 polarization) or IL-4 + anti-IFN gamma (classical Th2 polarization) for three days, and then were collected and detected for type I/type II cytokines by RT-PCR method. RESULTS: NK cells were purified from 15 healthy donors, over 70% purity of NK cells were determined by flow cytometry. NK cells in peripheral blood expressed high level of type I cytokines, mainly IFN gamma, but low level of type II cytokines such as IL-10 and IL-13, IL-4 was not produced by NK cells. Cells cultured in IFN gamma + anti-IL-4 condition exhibited significantly increased level of IFN gamma, unchanged IL-2, and decreased type II cytokines. Cells grew in IL-4 + anti-IFN gamma condition exhibited increased IL-10 and IL-13, and decreased IFN gamma expressions. CONCLUSIONS: Based on the cytokine production, NK cells may be divided into two functional subsets in the same manner as that of T lymphocytes(e.g. Th1/Th2): NKh1 and NKh2. The biological characterization and phenotypic marker are under investigate.

Effects of growth hormone and prolactin on hematopoiesis.

The use of the neuroendocrine hormones growth hormone (GH) and prolactin (PRL) in preclinical models, demonstrating promotion of hematopoietic recovery and immune function, offers promise for several clinical situations. These hormones do not appear to produce the same extent of immune/hematopoietic effects when compared to conventional hematopoietic and immune stimulating cytokines (i.e. G-CSF or interleukin-2). However, their pleiotropic effects and limited toxicity after systemic administration makes them attractive to test in myeloablative situations. More work needs to be performed to understand the mechanism(s) of GH and PRL action, particularly with regard to hematopoietic progenitor cell expansion and differentiation both in normal and pathologic situations.

Prolactin exerts hematopoietic growth-promoting effects in vivo and partially counteracts myelosuppression by azidothymidine.

Prolactin (PRL) is a neuroendocrine hormone that influences immune and hematopoietic development. The mechanism of action of this hormone in vivo remains unclear; therefore, we assessed the effects of PRL on hematopoiesis in vivo and in vitro. Normal resting mice were treated with 0, 1, 10, or 100 microg of recombinant human prolactin (rhPRL) for 4 consecutive days and euthanized on the fifth day for analysis of myeloid and erythroid progenitors in the bone marrow and spleen. Both frequencies and absolute numbers of splenic colony-forming unit granulocyte-macrophage (CFU-GM) and burst-forming unit-erythroid (BFU-e) were significantly increased in mice receiving rhPRL compared to the controls that had received saline only. Bone marrow cellularities were not significantly affected by any dose of rhPRL, but the absolute numbers and frequencies of bone marrow CFU-GM and BFU-e were augmented by rhPRL. These results suggest that rhPRL can promote hematopoiesis in vivo. Because rhPRL augments myeloid development in vivo, we examined the potential of the hormone to reverse the anemia and myelosuppression induced by azidothymidine (AZT). Mice were given rhPRL injections concurrent with 2.5 mg/mL AZT in drinking water. rhPRL partially restored hematocrits in the animals after 2 weeks of treatment and increased CFU-GM and BFU-e in both spleens and bone marrow. The experiments with AZT and rhPRL support the conclusion that the hormone increases myeloid and erythroid progenitor numbers in vivo, and they suggest that the hormone is clinically useful in reversing myelosuppression induced by AZT or other myeloablative therapies.

Suppression of graft-versus-host disease and amplification of graft-versus-tumor effects by activated natural killer cells after allogeneic bone marrow transplantation.

Bone marrow transplantation (BMT) is currently used for the treatment of a variety of neoplastic diseases. However, significant obstacles limiting the efficacy of allogeneic BMT are the occurrence of graft-versus-host disease (GvHD) and tumor relapse. Natural killer (NK) cells exert a variety of immunologic and homoeostatic functions. We examined whether adoptive transfer of activated NK cells of donor type would prevent GvHD after allogeneic BMT in mice. Lethally irradiated C57BL/6 (H-2(b)) mice, were transplanted with MHC incompatible BALB/c (H-2(d)) bone marrow cells and spleen cells and rapidly succumbed to acute GvHD. In contrast, mice that also received activated NK cells of donor type exhibited significant increases in survival. In determining the mechanism by which the NK cells prevented GvHD, mice were concurrently treated with a neutralizing antibodies to the immunosuppressive cytokine TGFbeta. Anti-TGFbeta completely abrogated the protective effects of the activated donor NK cells indicating that TGFbeta plays an important role in the prevention of GvHD by NK cells. We then examined whether activated NK cells of donor type after allogeneic BMT would induce graft-versus-tumor (GvT) effects without GvHD in mice bearing a murine colon adenocarcinoma (MCA-38). 10 d after receiving the tumor, in which the mice had demonstrable lung metastases, recipients received an allogeneic BMT with or without activated NK cells. Administration of activated NK cells resulted in significant GvT effects after allogeneic BMT as evidenced by increases in median survival and fewer lung metastasis. No evidence of GVHD was detected compared with recipients receiving spleen cells alone which also developed fewer lung metastases but in which all had succumbed to GVHD. Thus, our findings suggest that adoptive immunotherapy using activated donor NK cells combined with allogeneic BMT inhibits GvHD and promotes GvT in advanced tumor-bearing mice. These results also suggest that GvT and GvHD can be dissociable phenomena.

Recombinant human growth hormone promotes hematopoietic reconstitution after syngeneic bone marrow transplantation in mice.

Recombinant human growth hormone (rhGH) was administered to mice after syngeneic bone marrow transplantation (BMT) to determine its effect on hematopoietic reconstitution. BALB/c mice were given 10 microg intraperitoneal injections of rhGH every other day for a total of 10 injections following syngeneic BMT. Mice that received rhGH exhibited significant increases in total hematopoietic progenitor cell content (colony-forming unit-culture) in both bone marrow and spleen. Erythroid cell progenitor content (burst-forming unit-erythroid) was also significantly increased after rhGH treatment. Analysis of peripheral blood indicated that administration of rhGH resulted in significant increases in the rate of white blood cell and platelet recovery. Granulocyte marker 8C5+ cells were also increased in the bone marrow and spleens of treated mice. Red blood cell, hematocrit, and hemoglobin levels were increased at all time points after rhGH treatment. No significant pathologic effects or weight gain were observed in mice receiving repeated injections of 10 microg rhGH. Thus, rhGH administration after syngeneic BMT promoted multilineage hematopoietic reconstitution and may be of clinical use for accelerating hematopoiesis after autologous BMT.