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In this study, the spatiotemporal variation in carbon stocks under different vegetation coverages during the period 2001-2020 in Henan Province, China, was evaluated using the InVEST, PLUS, and geodetector models. The vegetation coverage patterns and corresponding carbon stocks were predicted under the three scenarios of natural trend, ecological degradation, and ecological restoration, the results show that: (1) Henan Province's vegetation area has been trending downward over the last 20 years, with losses in grassland, savanna, and farming vegetation. (2) The carbon stocks exhibited a low distribution pattern in the central and eastern parts of the country and a high distribution pattern in the northwest and southeast. The growth of total carbon stocks slowed. (3) The overall carbon stores exhibited a declining pattern throughout the three ecological scenarios. The carbon stocks experienced the least decline under the ecological restoration scenario. (4) Evapotranspiration, altitude, slope, distance from farmland and temperature were the main factors affecting the spatial differentiation of regional carbon stocks. Our results can aid in the sustainable management of ecosystems in Henan Province and provides a scientific basis and research concepts for the effective increase in carbon stocks. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Peripheral nerve block can provide effective postoperative analgesia to patients undergoing total hip arthroplasty (THA). This study aimed to compare ultrasound-guided pericapsular nerve group (PENG) block against anterior quadratus lumborum (AQL) block for pain management in primary THA. PATIENTS AND METHODS: In this prospective, double-blind, randomized controlled trial, 90 patients undergoing primary THA under general anesthesia were randomly allocated to receive ultrasound-guided PENG block + sham AQL block ("PENG group") or ultrasound-guided AQL block + sham PENG block ("AQL" group). The primary outcome was the highest pain score on a visual analogue scale while the patient was in the recovery room. Secondary outcomes included pain scores after transfer out of the recovery room, morphine consumption, quadricep strength, duration of hospitalization, pain level one year after surgery, and incidence of complications. RESULTS: Patients in the PENG group reported significantly lower maximum pain scores in the recovery room (31.3±9.1 vs. 37.3±7.4, p=0.001), as well as significantly lower pain scores at rest at 3 h after surgery and during motion at 3 and 6 h after surgery. The two groups did not differ significantly in postoperative morphine consumption, length of hospitalization, pain level at one year after surgery, or incidence of complications. Neither block significantly weakened the quadriceps. CONCLUSIONS: PENG block may provide slightly more effective postoperative analgesia than AQL block during the early recovery period after primary THA. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The aim of our study was to determine whether abnormal hyperplasia of chondrocytes occurs in glucocorticoid-induced osteonecrosis of the femoral head (GC-ONFH) using a well-established rat model. MATERIALS AND METHODS: Rats were injected with lipopolysaccharide and methylprednisolone to induce GC-ONFH, while control animals were injected with saline (12 animals per group). Establishment of the disease model was confirmed using micro-computed tomography and hematoxylin-eosin (HE) staining of femoral head tissue sections. Chondrocyte hyperplasia was detected using HE staining and semi-quantitated using toluidine blue and saffron O staining. Expression of the autophagy marker LC3B was assessed in cartilage tissues of femoral head using immunohistochemistry. RESULTS: G C-ONFH a nimals s howed s ignificantly greater area of abnormal chondrocyte hyperplasia in femoral head tissue sections than control animals. They also showed significantly higher expression of LC3B in articular cartilage of the femoral head. CONCLUSIONS: GC-ONFH may be associated with abnormal chondrocyte hyperplasia in articular surface cartilage, which may be related to glucocorticoid-induced overactivation of autophagy. [ABSTRACT FROM AUTHOR]

We have reported that murine bone marrow endothelial cells (mBMECI expressed thymosin [Beta]4 (T[Beta]4). In this report, the expression level of T[Beta]4 in mBMEC in different proliferation state was investigated. RT-PCR was used to detect the expression of T[Beta]4 mRNA and HPLC was used to determine the concentration of T[Beta]4 in bone marrow endothelial cell-conditioned medium (BMEC-CM). BMEC were cultured in three different culture media: IMDM only, routine culture medium, and IMDM + serum-supplemented bFGF. After incubation for 36 h, the cells were collected for the assay. BMEC-CM was collected from the confluent BMEC layer that was incubated for an additional 48 h without added serum or cytokines. The results showed that: (1) the cell number of BMEC in serum-free medium was the least while the cell number in bFGF-supplemented culture medium was the most (P [is less than] 0.05). [[sup.3]H]Thymidine incorporation showed a similar result (P [is less than] 0.05). (2) The relative level of T[Beta]4 mRNA of BMEC in three culture media was 8.16, 13.26, and 17.49%, respectively. The concentration of T[Beta]4 in BMEC-CM which the cells were treated by bFGF was much higher than in routine BMEC-CM (P [is less than] 0.01). (3) The expansion of the hematopoietic high-proliferation potential colony-forming cell (HPP-CFC) was 10.58 [+ or -] 1.32-fold in BMEC-CM from bFGF-treated cells, while in routine BMEC-CM was 6.09 [+ or -] 1.02-fold (P [is less than] 0.011 after 24 h of culturing. T[Beta]4 antiserum partially canceled the expansive effect. These results indicated that the expression level of T[Beta]4 was related to the proliferation state of BMEC. High expression of T[Beta]4 in BMEC may have the role of protecting the primitive hematopoietic cells from extensive differentiation.

Robot's blind tracking of an unknown surface with high speed is a critical problem of force control, to avoid damage to breakable objects, such as a bulb. It must deal with a typical action-based intelligence directly from force sensing to action in robot tracking and understanding of unknown external environments. Bio-inspired by human finger tracking, the realization of high-speed tracking relies on the real-time understanding and prediction of surface tendency, including the geometric feature and roughness. This paper presents an action-based intelligent frame for robot to track and understand an unknown surface with high speed using force sensing and control. The moving frame of force and position parallel control is built to obtain the differential geometric properties of an unknown surface. The motion trajectory is dynamically predicted by force sensing while tracking an unknown surface. A hybrid interpolator is developed to realize the parallel control of force and position. Moreover, a discriminant method for basic surface type recognition is investigated based on the normal curvature and geodesic torsion directly derived from the moving frame. The surface of revolution is further identified in the global understanding of unknown surface. The results of tracking a bulb with a desired contact force at a speed of 25 mm/s show the effectiveness of the proposed intelligent frame. [ABSTRACT FROM PUBLISHER]

For all solutions of a class of second order nonlinear damped differential equations, new oscillation criteria are established. Asympotic behavior for forced equations is also discussed. [ABSTRACT FROM AUTHOR]

A retrospective analysis of the clinical data of seven acute B-lymphoblastic leukemia (B-ALL) patients with TCF3-HLF fusion gene-positive admitted to the First Affiliated Hospital of Soochow University from June 2017 to August 2022 was conducted to summarize their clinical features and prognoses. The seven B-ALL patients comprised four males and three females, with a median age of 18 (11-33) years. Five patients tested positive for CD33 expression, and four patients had a normal karyotype. Two patients had hypercalcemia at the initial diagnosis, and one patient developed hypercalcemia at relapse. Six patients presented with coagulation dysfunction at diagnosis. After induction chemotherapy, five out of seven patients achieved complete remission, of which four subsequently relapsed. Two patients did not achieve remission even after two rounds of induction chemotherapy, with one achieving complete remission after treatment with blinatumomab immunotherapy. Three patients underwent chimeric antigen receptor T cell therapy, whereas three patients subsequently underwent hematopoietic stem cell transplantation. Five patients died, while two patients survived with sustained complete remission. TCF3-HLF-positive B-ALL is rare and has a high relapse rate and poor prognosis.

Objective: To compare the patient-reported outcomes and short-term clinical outcomes between robotic-assisted and laparoscopic-assisted radical gastrectomy for locally advanced gastric cancer. Methods: This single-center prospective randomized controlled trial was conducted in the Department of Gastrointestinal Surgery,Affiliated Hospital of Qingdao University from October 2020 to August 2022. Patients with locally advanced gastric cancer who were to undergo radical gastrectomy were selected and randomly divided into two groups according to 1∶1, and received robotic surgery and laparoscopic surgery, respectively. Patient-reported outcomes and short-term clinical outcomes (including postoperative complications, surgical quality and postoperative short-term recovery) were compared between the two groups by t test, Mann-Whitney U test, repeated ANOVA, generalized estimating equation, χ 2 test and Fisher's exact test. Results: A total of 237 patients were enrolled for modified intention-to-treat analysis (120 patients in the robotic group, 117 patients in the laparoscopic group). There were 180 males and 59 females, aged (63.0±10.2) years (range: 30 to 85 years). The incidence of postoperative complications was similar between the robotic group and laparoscopic group (16.7% (20/120) vs. 15.4% (18/117), χ 2 =0.072, P =0.788). The robotic group had higher patient-reported outcomes scores in general health status, emotional, and social domains compared to the laparoscopic group, differences in time effect, intervention effect, and interaction effect were statistically significant (general health status: χ 2 value were 275.68, 3.91, 6.38, P value were <0.01, 0.048, 0.041; emotional: χ 2 value were 77.79, 6.04, 6.15, P value were <0.01, 0.014, 0.046; social: χ 2 value were 148.00, 7.57, 5.98, P value were <0.01, 0.006, 0.048). However, the financial burden of the robotic group was higher, the differences in time effect, intervention effect and interaction effect were statistically significant ( χ 2 value were 156.24, 4.08, 36.56, P value were<0.01, 0.043,<0.01). Conclusion: Compared to the laparoscopic group, the robotic group could more effectively relieve postoperative negative emotions and improve recovery of social function in patients.

Objective: To investigate the clinical and laboratorial characteristics of patients with myelodysplastic syndrome (MDS) and erythroid hyperplasia. Methods: MDS patients whose bone marrow was hypercellular with erythroid lineage more than 50% and blasts account for less than 20% of non-erythroid cells were enrolled in this study. The ratio of mature erythrocytes to nucleated erythrocytes was no more than 20, namely MDS patients with erythroid hyperplasia(MDS-E). The retrospective analysis comprised 102 patients with MDS-E from the First Affiliated Hospital of Suzhou University. Clinical characteristics, karyotype, and the prognostic significance of erythroid hyperplasia were evaluated. Results: A total of 48 MDS-E patients (47.1%) presented a variety of cytogenetic abnormalities. The most frequently involved chromosomes were chromosome 8 (39.5% of all abnormal karyotypes), chromosome 7 (22.9%), followed by chromosome 5 (18.8%), chromosome 1 (16.7%) and chromosome 20 (16.7%). Hemoglobin (Hb) level affected the prognosis by survival analysis. The overall survival (OS) of MDS-E patients with Hb equal or more than 70 g/L was longer than that of patients less than 70 g/L ( P <0.001). Allogeneic hematopoietic stem cell transplantation (HSCT) significantly improved the OS compared with best supportive care ( P <0.001) and chemotherapy ( P <0.001). The extent of erythroid hyperplasia in bone marrow did not impact on prognosis ( P =0.187). Conclusions: Compared with previous reports of MDS patients, MDS-E patients have higher level of erythroid hyperplasia, more common erythroid dyshematopoiesis, more frequent 8 and 1 chromosome abnormalities. The degree of erythroid hyperplasia is not correlated with prognosis. Allogeneic hematopoietic stem cell transplantation improves the prognosis.

Objective: To investigate EVI1 expression and its associated clinical and cytogenetic characteristics in 447 acute myeloid leukemia (AML) patients. Methods: EVI1 expressions were measured in 447 AML cases from Jan. 2007 to Apr. 2015 to couple with clinical, cytogenetic and mutations' characteristics to summarize the features of AMLs with high EVI1 expression. Results: 17.9% of AML were high EVI1 expression (EVI1 + ), and the remainder low EVI1 expression (EVI1 - ). No significant differences between the two groups in terms of age, sex, hemoglobin level, white blood cell count and platelet count were observed. More M 0 , M 5 and M 6 subtypes were observed in EVI1 + group ( P = 0.027, 0.004 and 0.011, respectively). Cytogenetic abnormalities of 11q15, 11q23/MLL, 3q26, -7/7q- and t (9;11) were observed more frequently in EVI1 + group ( P <0.001, <0.001, <0.001, <0.001, =0.014, respectively). Normal karyotype, inv (16), t (8;21) were observed more frequent in EVI1 - group ( P =0.001, 0.009, 0.002, respectively). EVI1 + was more observed in high risk cytogenetics. Mutation of NPM1 was more observed in EVI1 - group ( P <0.001). Remission rate in EVI1 + group was significantly lower than EVI1 - group ( P <0.001). Leukemia-free survival was improved in EVI1 + AML patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Conclusions: High EVI1 expression was more observed in FAB subgroup M 5 , harbored more cytogenetic abnormalities of 11p15, 11q23/MLL, 3q26 rearrangement, -7/7q- and t (9;11). Remission rate of high EVI1 expression AML was lower, which could be improved by allo-HSCT.

The complete mitochondrial genome of the Southern catfish (Silurus meridionalis) and the Chinese catfish (S. asotus), was determined using the long and accurate polymerase chain reaction (LA-PCR) method. The mitochondrial DNA nucleotide sequences of S. meridionalis and S. asotus were compared with those of 47 other catfish species in the same order. The total length of mitochondrial DNA for S. meridionalis and S. asotus was 16,526 and 16,525 bp, respectively, and included 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a non-coding control region. This mitochondrial gene arrangement is identical to that observed in other Siluriformes. To determine the relative phylogenetic positions of S. meridionalis and S. asotus, and to discover phylogenetic relationships among 24 families of Siluriformes, analyses were conducted, based on mitochondrial DNA, 12S ribosomal RNA, 16S ribosomal RNA, and 13 protein-coding gene sequence data sets. Phylogenetic analyses were congruent with a basal split of the order into Clupeiformes, Characiformes, Cypriniformes, and Siluriformes, and supported a closer relationship of the Southern catfish (family Siluridae) and the Chinese catfish (family Siluridae) to Pimelodidae than to Bagridae. We concluded that these two species are part of a molecular clade that is different from that proposed in recent studies, in which Amblycipitidae appears as a sister group. Our results showed Amblycipitidae appearing as the most basal extant, and Bagridae appearing as a sister group of Cranoglanididae and Pangasiidae. The Siluriformes showed close phylogenetic relationship to the Characiformes.

Overcoming immune tolerance of the growth factors associated with tumor growth should be a useful approach to cancer therapy by active immunity. We used vascular endothelial growth factor (VEGF) as a model antigen to explore the feasibility of the immunogene tumor therapy with a vaccine based on a single xenogeneic homologous gene, targeting the growth factors associated with angiogenesis. To test this concept, we constructed a plasmid DNA encoding Xenopus homologous VEGF (XVEGF-p) and control vectors. We found that immunogene tumor therapy with a vaccine based on XVEGF was effective at both protective and therapeutic antitumor immunity in several tumor models in mice. VEGF-specific autoantibodies in sera of mice immunized with XVEGF-p could be found in Western blotting analysis and ELISA assay. The purified immunoglobulins were effective at the inhibition of VEGF-mediated endothelial cell proliferation in vitro, and at antitumor activity and the inhibition of angiogenesis by adoptive transfer in vivo. The elevation of VEGF in the sera of the tumor-bearing mice could be abrogated with XVEGF-p immunization. The antitumor activity and production of VEGF-specific autoantibodies, significantly elevated IgG1 and IgG2b, could be abrogated by the depletion of CD4(+) T lymphocytes. The observations may provide a vaccine strategy for cancer therapy through the induction of autoimmunity against the growth factors associated with tumor growth in a cross reaction with single xenogeneic homologous gene and may be of importance in the further exploration of the applications of other xenogeneic homologous genes identified in human and other animal genome sequence projects in cancer therapy.

In the present study, the effects of murine bone marrow endothelial cell conditioned medium (ECM) combined with flt3 ligand (FL) or/and thrombopoietin (TPO) on the proliferation of HPP-CFC and CFU-GM were investigated. Both ECM and the concentrated retentate of ECM (MW>10 kD) promoted the growth of CFU-GM and HPP-CFC, and this promoting effect was further enhanced by addition of FL or TPO. Using reverse transcriptase-polymerase chain reaction (RT-PCR) technique, the expression of FL and TPO mRNA was not found in murine bone marrow endothelial cells.

The title compound, 2,2'-(2,4,8,10-tetrathiaspiro[5.5]undecane-3,9-diylidene)bis(propanedinitrile), C(13)H(8)N(4)S(4), has been designed and synthesized for use as a potential new organic molecular electronic material. The spiro-annulated structure has twofold symmetry and is formed by two equal push-pull ethylene units, with the cycloalkylthio groups as electron donors and the cyano groups as electron acceptors. The intermolecular S.N non-bonded separation within a layer in the lattice is 3.296 (6) A, indicating a strong intermolecular interaction between the cyano groups and the S atoms, while the S atoms in two neighbouring molecules have a shortest S.S contact of 3.449 (3) A. In addition, attractive C-H.N and C-H.S interactions bridge adjacent molecules either within a layer or between layers. In short, these four types of intermolecular interactions combine to form an extended three-dimensional network in the lattice, resulting in a highly ordered array of molecular packing.

Aim: To study the effects of cimetidine (Cim) on the production of interferon gamma (IFN gamma) and tumor necrosis factor alpha (TNF alpha) by splenocytes in immune-derived aplastic anemic (AA) mice., Methods: Aplastic anemic mice model was constructed first, and then the splenocytes were induced to secrete IFN gamma and TNF alpha. Concentration of IFN gamma was assayed using sandwich ELISA, while that of TNF alpha was measured with L929 cytotoxicity methods., Results: (1) Concentrations of IFN gamma and TNF alpha secreted by splenocytes from AA mice were (137 +/- 36) ng/L and (6 +/- 3) microg/L, respectively, much more than the irradiated and the control mice. (2) Treatment with Cim 10 micromol/L reduced the concentrations of IFN gamma and TNF alpha to (14 +/- 8) ng/L and (2.7 +/- 0.6) microg/L, respectively., Conclusion: Cim could effectively reduce the production of IFN gamma and TNF alpha from splenocytes of AA mice.

The effects of > 10 kD component of serum-free conditioned medium from murine bone marrow-derived endothelial cell line on the formation of CFU-GM and HPP-CFC were investigated. We found that > 10 kD component alone could stimulate CFU-GM colony formation in dose-dependent. Above- 10 kD component supplemented with GM-CSF not only resulted in significant increase of CFU-GM as compared with > 10 kD or GM-CSF alone, but also could stimulate HPP-CFC formation. The effect of > 10 kD component on the formation of HPP-CFC was also in dose-dependent. The component above- 10 kD combined with IL-3, IL-6, EPO, SCF or GM-CSF further enhanced HPP-CFC colony formation. The results suggest that > 10 kD component may be used as the substitutes of hematopoietic growth factors to stimulate the formation of CFU-GM and HPP-CFC.

By using CFU-GM/CFU-L colony assay, NBT/MTT reductant test and DNA fragmentation analysis, we studied the effects of Sophora flavescens (SF) on CFU-GM proliferative ratio in human normal bone marrow/umbilical cord blood and on proliferation, differentiation and apoptosis in human acute myelogenous leukemia HL-60 cells. The results showed that 5, 10, 15, 20 micrograms.microliter-1 of SF significantly inhibited proliferation and induced apoptosis in the HL-60 cells in a dose-dependent fashion. Fifteen micrograms.microliter-1 of SF also induced differentiation in HL-60 cells. Furthermore, cytotoxic activity of SF(5-15 micrograms.microliter-1) was not apparent on human normal hematopoietic progenitors(CFU-GM). The results indicate that an appropriate concentration of SF has a selective antileukemic effect. Thus, these are important impetuses for further research of SF as an anticancer agent.

The breaking of immune tolerance against autologous angiogenic endothelial cells should be a useful approach for cancer therapy. Here we show that immunotherapy of tumors using fixed xenogeneic whole endothelial cells as a vaccine was effective in affording protection from tumor growth, inducing regression of established tumors and prolonging survival of tumor-bearing mice. Furthermore, autoreactive immunity targeting to microvessels in solid tumors was induced and was probably responsible for the anti-tumor activity. These observations may provide a new vaccine strategy for cancer therapy through the induction of an autoimmune response against the tumor endothelium in a cross-reaction.

Recently, cytokines and interleukins such as SCF, GM-CSF, G-CSF, TGF-beta, IL-6, IL-7, IL-8, IL-11 have been reported to be elaborated by endothelial cells. For further study, serum free bone marrow endothelial cell conditioned medium (BMEC-CM) was collected and ultrafiltrated by using a centriprep 10. The concentrated retentate (R-BMEC-CM) contained some substances whose molecular weight was more than 10 000 daltons. The filtrate (F-BMEC-CM) contained some substances whose molecular weight was less than 10 000 daltons. The effects of R-BMEC-CM and F-BMEC-CM on the growth of haematopoietic progenitors and the expression of cytokine and interleukin mRNAs of BMEC were investigated. The results showed that R-BMEC-CM stimulated the growth of CFU-GM, HPP-CFC, BFU-E, CFU-E, and CFU-Meg; while F-BMEC-CM inhibited the growth of these progenitors. Using the method of hybridizing to the Atlas cDNA Array, we were able to detect the presence of mRNAs of cytokines and interleukins in bone marrow endothelial cells. Our finding of the existence of mRNAs of SCF, GM-CSF, IL-6, TGF-beta, IL-1, and IL-11 in these cells was in agreement with the data reported previously. Furthermore, we detected mRNAs of MIP-2, Thymosion-beta4, PDGF, MSP-1, IFN-gamma, IL-13 and inhibin, which are related to haematopoiesis. Among these cytokines and interleukins, SCF, GM-CSF, IL-6, IL-1, and IL-11 are haematopoietic stimulators which may be responsible for the stimulative effects on the growth of haematopoietic progenitors. One of our new findings, the thymosin-beta4, is a small molecular haematopoietic inhibitor. It may be responsible for the inhibitory effect of F-BMEC-CM on haematopoietic progenitors. The presence of mRNAs of BMP, MSP-1, MIP-2, PDGF and IL-13 suggests that bone marrow endothelial cells might elaborate these substances. Their influence on haematopoietic progenitors needs further study., (Copyright 2000 Academic Press.)

Objective: To evaluate the effect of curcumin on the proliferation of hematopoietic progenitor cells or leukemic stem cells., Methods: Mouse bone marrow cells (colony forming unit-granulocyte and macrophage, CFU-GM) and WEHI-3B cells were observed using the colony assays., Results: The curcumin significantly inhibited the proliferation of both CFU-GM and WEHI-3B cells in a dose-dependent manner ranging from 10(-8) to 10(-4) mol.L-1; their IC50S were 1.036 x 10(-5) mol.L-1 and 1.220 x 10(-6) mol.L-1 of curcumin respectively., Conclusion: The proliferation of murine CFU-GM and WEHI-3B cells can be suppressed by curcumin. The inhibitory effect of curcumin on the proliferation of WEHI-3B cells is stronger than that of CFU-GM.

Serum-free murine bone marrow endothelial cell conditioned medium(mBMEC-cm) was collected. The mBMEC-cm was ultrafiltered in Centriprep. The concentrated retentive substance of mBMEC-cm (MW > 10,000) and filtrate of mBMEC-cm(MW < 10,000) were obtained. The effects of mBMEC-cm on the growth of CFU-E and BFU-E were investigated. The results showed: 1. The retentive substance of mBMEC-cm significantly enhanced the growth of CFU-E and BFU-E. The effects of mBMEC-cm(2%-6%, V/V) on the growth of CFU-E and BFU-E were dose-dependent. 2. The filtrate of mBMEC-cm markedly inhibited the growth of CFU-E and BFU-E. The effects of the filtrate of mBMEC-cm on the growth of CFU-E and BFU-E were also dose-dependent. The mechanism of regulation will be studied further.

In this study, the effects of bone marrow endothelial cell conditioned medium (BMEC-CM) on hematopoietic progenitors and the expression of cytokines mRNA of BMEC were investigated. BMEC-CM was ultrafiltrated in Centriprep-10, an ultrafiltering concentrator. The retentate (MW>10 kD) and filtrate (MW<10 kD) were obtained. The effects of different components of BMEC-CM on the murine hematopoietic progenitors were examined. The results showed that the retentate of BMEC-CM (MW>10 kD) significantly promoted the growth of CFU-GM, CFU-E, BFU-E, CFU-Meg and HPP-CFC, while the growth of CFU-GM, CFU-E, BFU-E, CFU-Meg and HPP-CFC were markedly inhibited by the filtrate of BMEC-CM (MW<10 kD). The effects of BMEC-CM were dose-dependent. The presence of mRNAs for GM-CSF, TGF-beta, bone morphogenetic protein 2A, basic fibroblast growth factor receptor, endothelin-2, macrophage-stimulating 1 (MSP-1), thymosin-beta10, connective tissue growth factor, macrophage inflammatory protein 2 alpha (MIP-2alpha), IL-6, IL-11, IL-13 and IFN-gamma mRNA were successfully detected by Atlas Arrays.

In this study, we obtained the serum-free conditioned medium from subcultures of murine bone-marrow-derived endothelial cell line which has been established recently by ourselves. And then, molecular weight (MW) > 10 kD, 3-10 kD, 1-3 kD, 0.5-1 kD and < 0.05 kD components were sifted out from the conditioned medium (mBMEC-CM) by means of serial ultrafiltration. Assays of granulocyte-macrophage colony-forming cell (CFU-GM) were performed to examine the effects of mBMEC-CM and its ultrafiltration-prepared components. It was not observed that mBMEC-CM and MW 3-10 kD component had any significant influence on the growth of CFU-GM. However, MW > 10 kD and 0.5-1 kD components enhanced the proliferation of CFU-GM, whereas MW 1-3 kD and < 0.5 kD ones inhibited it. All these four components exerted their effects on the growth of CFU-GM in dose-dependent way. Our observations suggest that under the conditions of in vitro, murine bone marrow endothelial cells produce several active components which have promoting or inhibitory effects on the growth of CFU-GM.

Stromal cells are believed to regulate hematopoiesis through direct cell-cell contact interactions and the release of growth factors. Many questions remain, however, about their lineage derivation and functional heterogeneity. We have previously shown that the adherent nontransformed, nonimmortalized murine bone marrow stromal cell population consists of three cell types which could be grown separately in vitro. Based on the phenotype characterization and expression of surface antigens, we proposed a classification listing for murine bone marrow stromal cells as macrophages, endothelial-like cells and myofibroblasts that display smooth muscle-like characteristics in culture. The present study describes the ability of each of these freshly isolated separated murine stromal cell populations to support the growth of primitive hematopoietic stem cells previously characterized as highly enriched in long-term repopulating cells (LTRC). Of the three stromal cell types tested only the myofibroblasts were capable of support for multilineage hematopoiesis derived in vitro from LTRC in a cloning ring culture system. Endothelial-like cells had an inhibitory effect on the proliferation of LTRC and their descendant cells that was induced by exogenous growth factors. This inhibitory activity was present in a low molecular weight filtrate of endothelial-like cells culture medium. This suggests an essential role for marrow stroma myofibroblasts in the support of proliferation of hematopoietic cells at the stage of early divisions of primitive hematopoietic stem cells and endothelial-like cells as negative regulators of this proliferation.

The passages 0 and 3 purified human bone marrow fibroblast layers (FLs) were established by long-term liquid cultures. After 12-day coculture of stromal. cell-depleted marrow cell suspensions with passage 0 FLs, 68.33 +/- 4.04% of the hemopoietic colonies adhered to FLs was myeloid in nature and the other 31.67 +/- 4.04% was erythroid. There were still CFU-E (colony forming unit-erythroid), BFU-E (burst forming unit- erythroid), and CFU-GM (colony forming unit-granulocyte/macrophage) among the nonadherent cells. The media conditioned by passage 0 (F0-CM) and passage 3(F3-CM) FLs stimulated the growth of myeloid and erythroid (BFU-E) colonies. From these data, it is concluded that both FLs and the media conditioned by fibroblasts can stimulate myeloid and erythroid hemopoiesis.

Tb(III) was used as a fluorescent probe in the study of the calcium-binding sites on Bovine Serum Albumin (BSA). The fluorescence of Tb(III) is enhanced markedly when bound to BSA and nonradiative energy transfer between two fluorescent tryptophan(Trp) residues and Tb(III) bound to calcium-binding sites on BSA occurred. Experimental results show that the major groups in BSA bound to metal ion are the carboxyl side groups of glutamic acid (Glu) and aspartic acid (Asp). The average distance between the bound Tb(III) and the two tryptophan residues in BSA calculated by a Föster dipole-dipole nonradiative energy transfer mechanism is 1.48 nm.

The effects of several growth factors on the proliferation of fibroblastic colony-forming units (CFU-F) were studied. In the present study CFU-F colonies were found to consist of fibroblasts, macrophages, and endothelial cells. Growth factors, including interleukin 3 (IL-3), interleukin 1 alpha (IL-1 alpha), epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and buffalo rat liver cell-conditioned medium (BRL-CM) were tested for stimulation of the proliferation of CFU-F in a standard culture in both 2% and 15% serum. Overall, the colony numbers produced in 15% serum were much higher than in 2% serum with or without growth factors. However, the influence of several growth factors on CFU-F cultured in 2% serum was relatively greater than in 15% serum when compared to controls. The stimulation of CFU-F by FGF only occurred in culture with 15% serum, and the stimulation by PDGF only occurred with 2% serum. Overall, the strongest stimulations were produced by PDGF, IL-3, and BRL-CM. Combining the other growth factors with IL-3, PDGF, or IL-1 alpha enhanced their effects only modestly. The stimulation by growth factors included increases of the cell numbers between and within colonies as well as an increase in the number of colonies. The study produced results that suggest a complex interaction mediated by growth factors between fibroblasts and other stromal cells within the CFU-F colonies and within the bone marrow itself.

In this study it was found that three kinds of cells, fibroblasts, endothelial cells, and macrophages, exist in fibroblastic colony-forming unit (CFU-F) colonies, consistent with the findings of others. However, when low enough bone marrow stromal cell numbers were cultured per dish, pure fibroblast, endothelial, and macrophage colonies were observed. When the cell number cultured in each dish was increased, all colonies were of mixed cell type. This evidence strongly suggests that the macrophages and endothelial cells contained in CFU-F colonies come from locally stimulated growth resulting in contamination and not from a common progenitor with fibroblasts or each other. In some longer term experiments, adipocytes were seen to appear within the colonies of late-passage pure fibroblast cultures.