[Efficacy and Safety of Plerixafor Combined with G-CSF for Autologous Peripheral Blood Hematopoietic Stem Cell Mobilization in Lymphoma Patients].

OBJECTIVE: To investigate the efficacy and safety of plerixafor combined with granulocyte colony-stimulating factor (G-CSF) in mobilizing peripheral blood hematopoietic stem cells in patients with lymphoma. METHODS: The clinical data of lymphoma patients who received autologous hematopoietic stem cell mobilization using plerixafor combined with G-CSF from January 2019 to December 2021 were retrospectively analyzed. The patients received 3 kinds of mobilization regimens: front-line steady-state mobilization, preemptive intervention, and recuse mobilization. The acquisition success rate, excellent rate of collection, and incidence of treatment-related adverse reaction were counted. The influence of sex, age, disease remission status, bone marrow involvement at diagnosis, chemotherapy lines, number of chemotherapy, platelet count and number of CD34+ cells on the day before acquisition in peripheral blood on the collection results were analyzed to identify the risk factors associated with poor stem cell collection. RESULTS: A total of 43 patients with lymphoma were enrolled, including 7 cases who received front-line steady-state mobilization, 19 cases who received preemptive intervention, and 17 cases who received recuse mobilization. The overall acquisition success rate was 58.1% (25/43) after use of plerixafor combined with G-CSF, and acquisition success rate of front-line steady-state mobilization, preemptive intervention, and recuse mobilization was 100%, 57.9%(11/19), and 41.2%(7/17), respectively. The excellent rate of collection was 18.6%(8/43). A total of 15 patients experienced mild to moderate treatment-related adverse reactions. The number of CD34+ cells < 5 cells/µl in peripheral blood on the day before collection was an independent risk factor affecting stem cell collection. CONCLUSIONS: Plerixafor combined with G-CSF is a safe and effective mobilization regimen for patients with lymphoma. The number of CD34+ cells in peripheral blood on the day before collection is an predictable index for the evaluation of stem cell collection.

[Effects of intensive management on abundance and composition of soil N2-fixing bacteria in Phyllostachys heterocycla stands].

Denaturing gradient-gel electrophoresis and real-time quantitative PCR (qPCR) were employed to determine the effects of intensive management on soil N2-fixing bacteria in a moso bamboo (Phyllostachys heterocycla) plantation. Soil samples were collected from the moso bamboo stands receiving 0 (CK), 10, 15, 20, and 25 years of intensive management. It was found that intensive management caused a strong decrease in soil pH but a general increase in soil available nutrients. The structure of the N2-fixing bacterial communities in the soils having received 10 and 25 years of intensive management were quite similar to that from the CK; however, those from 15 and 20 years of intensification differed from the CK. With increasing time of intensive management, the abundance and diversity of the nifH gene at first decreased and then increased, with the minimum values being observed after 15 years of intensive management, indicating the eventual resiliency of N2-fixing bacteria to disturbance induced by intensive management. Redundancy analysis indicated that soil available potassium, available nitrogen, nitrite nitrogen, and ammonium nitrogen were more closely related to the changes of N2-fixing bacterial community structure compared with the other soil indices measured. In conclusion, the soil N2-fixing bacterial community was negatively affected by intensive management in the short term, but could recover in the long term.

Metformin displays anti-myeloma activity and synergistic effect with dexamethasone in in vitro and in vivo xenograft models.

Epidemiologic studies and meta-analyses have suggested that patients with type 2 diabetes mellitus (T2DM) have a higher incidence of malignancies, including myeloma. Metformin is a widely prescribed antidiabetic drug. Recently, researchers have shown that metformin has direct anticancer activity against many tumor cell lines, mainly through activating AMP-activated protein kinase (AMPK) or reducing the blood insulin level. In the present study, we investigated whether metformin exerts an anti-myeloma effect in in vitro and in vivo xenograft models and explored the underlying mechanism. We found that metformin can inhibit proliferation of MM cells by inducing apoptosis and cell cycle arrest in the G0/G1 phase. Western blot showed that metformin activated caspase 3, caspase 9, PARP-1, Bak, and p21 and inactivated Mcl-1, HIAP-1, cyclin D1, CDK4, and CDK6. Metformin inhibited the expression of insulin growth factor-I receptor (IGF-IR), and phosphatidyl inositol 3-kinase (PI3K), protein kinase B (PKB/AKT) and the downstream mammalian target of rapamycin (mTOR). IGF-I blocked metformin-induced MM cell apoptosis and reactivation of the PI3K/AKT/mTOR signaling pathway. Metformin also demonstrated synergistic activity with dexamethasone but not bortezomib to eradicate MM cells in vitro and in vivo, especially in MM.1S cells. We conclude that metformin inhibits MM cell proliferation through the IGF-1R/PI3K/AKT/mTOR signaling pathway. Metformin and dexamethasone combination therapy may be an option for MM treatment.

[Effects of bamboo charcoal on the growth of Trifolium repens and soil bacterial community structure].

The effects of addition rates (0, 3% and 9%) and particle sizes (0.05, 0.05-1.0 and 1.0-2.0 mm) of bamboo charcoal on the growth of Trifolium repens and soil microbial community structure were investigated. The results showed that bamboo charcoal addition greatly promoted the early growth of T. repens, with the 9% charcoal addition rate being slightly better than the 3% charcoal addition rate. The effects of different particle sizes of bamboo charcoal on the growth of T. repens were not different significantly. Growth promotion declined with time during 120 days after sowing, and disappeared completely after 5 months. DGGE analysis of the bacterial 16S rDNA V3 fragment indicated that bamboo charcoal altered the soil bacterial community structure. The amount and Shannon diversity index of bacteria in the bamboo charcoal addition treatments increased compared with CK. The quantitative analysis showed that the amount of bacteria in the treatment with bamboo charcoal of fine particle (D < 0.05 mm) at the 9% addition rate was significantly higher than in the other treatments. The fine bamboo charcoal had a great effect on soil bacteria amount compared with the charcoal of other sizes at the same addition rate.

Fibroblast activation protein protects bortezomib-induced apoptosis in multiple myeloma cells through ß-catenin signaling pathway.

Multiple myeloma (MM) is a malignant plasma cells proliferative disease. The intricate cross-talk of myeloma cells with bone marrow microenvironment plays an important role in facilitating growth and survival of myeloma cells. Bone marrow mesenchymal stem cells (BMMSCs) are important cells in MM microenvironment. In solid tumors, BMMSCs can be educated by tumor cells to become cancer-associated fibroblasts (CAFs) with high expression of fibroblast activation protein (FAP). FAP was reported to be involved in drug resistance, tumorigenesis, neoplastic progression, angiogenesis, invasion, and metastasis of tumor cells. However, the expression and the role of FAP in MM bone marrow microenvironment are still less known. The present study is aimed to investigate the expression of FAP, the role of FAP, and its relevant signaling pathway in regulating apoptosis induced by bortezomib in MM cells. In this study, our data illustrated that the expression levels of FAP were not different between the cultured BMMSCs isolated from MM patients and normal donors. The expression levels of FAP can be increased by tumor cells conditioned medium (TCCM) stimulation or coculture with RPMI8226 cells. FAP has important role in BMMSCs mediated protecting MM cell lines from apoptosis induced by bortezomib. Further study showed that this process may likely through ß-catenin signaling pathway in vitro. The activation of ß-catenin in MM cell lines was dependent on direct contact with BMMSCs other than separated by transwell or additional condition medium from BMMSCs and cytokines.

Poly[µ(2)-aqua-diaqua-(µ(8)-3-nitro-benzene-1,2-dicarboxylato)(µ(6)-3-nitro-benzene-1,2-dicarboxylato)tetra-sodium].

In the title layered coordination polymer, [Na(4)(C(8)H(3)NO(6))(2)(H(2)O)(3)](n), the doubly deprotonated 3-nitro-benzene-1,2-dicarboxyl-ate ligands exhibit µ(8)- and µ(6)-coordination modes to the sodium ions, generating sheets lying parallel to (001). The coordination environments of the sodium ions are distorted octa-hedral, distorted trigonal-bipyramidal and moncapped trigonal-prismatic. One of the nitro groups is disordered over two sets of sites with site-occupancy factors 0.580 (8):0.419 (2). A network of O-H⋯O and O-H⋯N hydrogen bonds helps to establish the packing.

Low-temperature Heat Capacities and Thermodynamic Properties of Crystalline 2-Aminopyridinium Benzoate (C12H12N2O2) (s).

2-Aminopyridinium benzoate was synthesized. Chemical analysis, elemental analysis, and X-ray crystallography were applied to characterize the composition and crystal structure of the compound. The lattice potential energy of the title compound was calculated to be UPOT = 284.297 kJ mol-1. Low-temperature heat capacities of the compound were measured by a precision automatic adiabatic calorimeter over the temperature range from 78 K to 365 K. A polynomial equation of heat capacities against the temperature in the region of 78 K to 365 K was fitted by a least square method. Based on the fitted polynomial equation, the smoothed heat capacities and thermodynamic functions of the compound relative to the standard reference temperature 298.15 K were calculated at intervals of 5 K. According to the synthesis reaction, the standard molar enthalpies of dissolution for the reactants and product in the selected solvent were measured by an isoperibol solution-reaction calorimeter, respectively. Accordingly, the enthalpy change of the synthesis reaction was calculated to be ΔrHom = -(20.016 ± 0.182) kJ mol-1. Finally, the standard molar enthalpy of formation of 2-aminopyridinium benzoate was determined to be ΔfHom = - (365.416 ± 0.961) kJ mol-1 in accordance with Hess law.

[Study on the relationship between the expression of SOCS-1/3 in the peripheral blood mononuclear cells in patients with multiple organ dysfunction syndrome and their prognosis].

OBJECTIVE: To observe the mRNA and protein expression of suppressor of cytokines signaling protein 1/3 (SOCS-1/3) in the peripheral blood mononuclear cells (PBMCs) in patients with multiple organ dysfunction syndrome (MODS), and to explore the relationship between the SOCS expression and the patients' prognosis. METHODS: Peripheral blood specimens of 32 patients with MODS and 24 healthy volunteers (controls) were collected and the PBMCs were isolated by centrifugation and Ficoll-Hypaque sedimentation. The mRNA of SOCS-1/3 was determined by reverse transcription-polymerase chain reaction (RT-PCR). And the protein content of SOCS-1/3 was determined by Western blotting. The relationship between the SOCS expression and the patients' prognosis was analyzed. RESULTS: There was no significant difference in the SOCS-1 mRNA expression between control group and MODS group (0.526+/-0.044 vs. 0.466+/-0.047, P>0.05). The SOCS-1 expression of MODS group was significantly higher than that of control group (0.814+/-0.045 vs. 0.479+/-0.021, P<0.05). In the MODS group, the SOCS-1 mRNA expression and protein content of dead patients were significantly lower than those of survived patients (mRNA 0.487+/-0.032 vs. 0.532+/-0.028, protein 0.787+/-0. 029 vs. 0.838+/-0.040, both P<0.05). There was significant negative correlation between the SOCS-1 mRNA expression and the MODS score (r1=-0.731,P<0.01). There was also significant negative correlation between the SOCS-1 content and the MODS score (r2=-0.526,P<0.01). The SOCS-3 mRNA expression of MODS group was higher than that of control group (0.993+/-0.415 vs. 0.461+/-0.046, P<0.05). The SOCS-3 content in the PBMCs of MODS groups was significant higher than that of control group (0.458+/-0.033 vs. 0.403+/-0. 024, P<0.05). In the MODS group, the SOCS-3 mRNA expression and protein content of dead patients were significant higher than those of survived patients (mRNA 1. 245+/-0.408 vs. 0.805+/-0.326, protein 0.486+/-0.021 vs. 0.425+/-0.016, both P<0.05). There was positive correlation between the SOCS-3 mRNA expression and the MODS score but the correlation was not significant (r=0. 468, P>0.05). And there was significant positive correlation between the SOCS-3 content and the MODS score (r=0.783, P<0.01). CONCLUSION: SOCS-1 may protect the tissue from further damage while SOCS-3 may cause tissue damage indirectly. The detection of SOCS-1/3 may help to predict the prognosis of MODS.

[Role of Bid protein in the mitochondria and Endoplasmic Reticulum associated apoptotic pathway].

OBJECTIVE: To explore the role of Bid protein in the mitochondria and endoplasmic reticulum (ER) associated apoptotic pathway. METHODS: Apoptosis of MUTZ-1 cells induced by homoharringtonine (HHT) was measured by FACS. Mitochondria and ER associated apoptotic pathway was detected by RT-PCR and Western blotting. And the translocation of Bid protein was measured by laser scanning confocal microscope (LSCM). RESULTS: After exposure of MUTZ-1 to HHT at 0.05 microg/ml for 24 h, typical ER-stress phenomenon induced apoptotic cells and release of Ca2+ from the cytosolic Ca2+ storage and the loss of mitochondrial membrane potential were observed. RT-PCR analysis revealed that mRNAs for ER stress-associated proapoptotic factor were markedly increased at 4 h after 0.05 microg/ml HHT treatment and peaked at 12 h, then decreased steady. Activation of caspase protein was also observed at 8 h. The translocation of Bid protein from ER to mitochondria was observed at 12 h after HHT treatment. CONCLUSION: HHT can induce MUTZ-1 cells apoptosis. The cell death may be likely mediated by the ER stress pathway as well as mitochondrial pathway and Bid protein may be the cross talk of the two apoptotic pathways.

[E1A gene transfection of human undifferentiated thyroid cancer cell line HTC/3 by nanoparticles].

OBJECTIVE: To prepare nanoparticles containing E1A gene and observe the efficiency and feasibility of transfecting E1A gene into human undifferentiated thyroid cancer cell line HTC/3. To examine the sensitivity of transgene cells to X-ray and X-ray-induced apoptosis in those cells. METHODS: Nanoparticle-DNA complex was prepared with PLGA coating adenoviral early expression gene E1A, and the package efficiency, release progress in vitro, and size of the complex were determined. The nanoparticle-DNA was transfected into the HTC/3 cells. Lipofectamine was used to transfect E1A gene as a control. RT-PCR was used to examine E1A gene mRNA expression in the transfected cells. The survival ratio of HTC/3-E1A and control cells, and the growth inhibition ratio induced by different doses of X-ray in HTC/3-E1A cells were examined by MTT assay. The apoptosis in HTC/3-E1A cells induced by 2 Gy X-ray iradiation was examined by flow cytometry and DNA electrophoresis. RESULTS: The package efficiency, release progress in vitro, and size of the nanoparticle-DNA complex were 0.78%, 18 days, and 150-280 nm, respectively when transfected the plasmid at the same level, the nanoparticle group got more positive transgene cell clones than that in lipofectamine group, with a statistically significant difference (P < 0.05). RT-PCR showed that transgenic cells from both nanoparticle-DNA and lipofectamine groups had E1A gene mRNA expression. The HTC/3-E1A cells grew slowly, and their doubling time was prolongated (1.44 times in comparison with that in parental cells). According to IC50, the sensitivity of HTC/3-E1A cells to X-ray was improved 2.9 and 2.8 times, respectively, in comparison with that in HTC/3-Vect and HTC/3 cells. The ratio of subG0/G1 phase of HTC/3-E1A cells was significantly higher than that in HTC/3-Vect and HTC/3 cells (P < 0.01). The ratio of S phase of HTC/3-E1A cells was significantly lower than that in HTC/3-Vect and HTC/3 cells (P < 0.01). A typical DNA ladder pattern of apoptosis in HTC/3-E1A cells was observed by electrophoresis, but not found in HTC/3-Vect and HTC/3 cells. CONCLUSION: A nanoparticle-DNA complex has been successfully prepared, and it may carry a foreign gene into cells. The sensitivity of HTC/3-E1A cells to X-ray is significantly improved. Moreover, apoptosis is induced by x-ray in the E1A gene-transfected cells.