Exosomes derived from mesenchymal stromal cells exert a therapeutic effect on hypoxia-induced pulmonary hypertension by modulating the YAP1/SPP1 signaling pathway.

OBJECTIVE: Hypoxic pulmonary hypertension (HPH) is a progressive and life-threatening disease characterized by perivascular inflammation, pulmonary vascular remodeling, and occlusion. Mesenchymal stromal cell-derived exosomes (MSC-exo) have emerged as potential therapeutic agents due to their role in cell communication and the transportation of bioactive molecules. In this study, we aimed to investigate the therapeutic effects of MSC-exo against HPH and elucidate the underlying molecular mechanism. METHODS: Exosomes were isolated from conditioned media of human bone mesenchymal stromal cells using ultracentrifugation and characterized through western blotting, transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). An HPH animal model was established in male SD rats, and MSC-exo or phosphate-buffered saline (PBS) were administered via the tail vein for three weeks. Subsequently, right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), and pulmonary vascular remodeling were evaluated. Lung tissues from HPH rats and normal rats underwent high-throughput sequencing and transcriptomic analysis. Gene Ontology (GO) analysis was employed to identify upregulated differentially expressed genes. Additionally, rat pulmonary artery smooth muscle cells (PASMC) exposed to platelet-derived growth factor-BB (PDGF-BB) were used to simulate HPH-related pathological behavior. In vitro cellular models were established to examine the molecular mechanism of MSC-exo in HPH. RESULTS: MSC-exo administration protected rats from hypoxia-induced increases in RVSP, RVHI, and pulmonary vascular remodeling. Additionally, MSC-exo alleviated PDGF-BB-induced proliferation and migration of PASMC. Transcriptomic analysis revealed 267 upregulated genes in lung tissues of HPH rats compared to control rats. Gene Ontology analysis indicated significant differences in pathways associated with Yes Associated Protein 1 (YAP1), a key regulator of cell proliferation and organ size. RT-qPCR and western blot analysis confirmed significantly increased expression of YAP1 in HPH lung tissues and PASMC, which was inhibited by MSC-exo treatment. Furthermore, analysis of datasets demonstrated that Secreted Phosphoprotein 1 (SPP1), also known as Osteopontin (OPN), is a downstream binding protein of YAP1 and can be upregulated by PDGF-BB. MSC-exo treatment reduced the expression of both YAP1 and SPP1. Lentivirus-mediated knockdown of YAP1 inhibited PDGF-BB-induced PASMC proliferation, migration, and SPP1 protein levels. CONCLUSION: Our findings demonstrate that MSC-exo exert a therapeutic effect against hypoxia-induced pulmonary hypertension by modulating the YAP1/SPP1 signaling pathway. The inhibition of YAP1 and downstream SPP1 expression by MSC-exo may contribute to the attenuation of pulmonary vascular remodeling and PASMC proliferation and migration. These results suggest that MSC-exo could serve as a potential therapeutic strategy for the treatment of HPH. Further investigations are warranted to explore the clinical applicability of MSC-exo-based therapies in HPH patients.

Regulating structural dimensionality and emission colors by organic conjugation between SmIII at a fixed distance.

The conjugation of bridging bis(diphenylphosphine oxide) alkane or arene ligands was found to control the structural dimensionality and the emission color of complexes from reactions with SmIII(hfac)3(H2O)2 (hfac- = hexafluoroacetylacetonato) while retaining the SmSm distances. Bis(diphenylphosphine oxide)-1,4-butane (L1) affords a one-dimensional (1D) ribbon {Sm(hfac)3(L1)}∞ (1) that emits red color, while bis(diphenyl-phosphinoyl)-1,4-benzene (L2) results in a two-dimensional (2D) network {Sm(hfac)2(CF3COO)(L2)3}∞ (2) and near-white emission, but bis(diphenyl-phosphinoyl)-9,10-anthracene (L3) forms a zero-dimensional (0D) cyclic structure {Sm(hfac)3(L3)}2 (3) with strong ππ interactions that emit green color. Noticeably, the conjugation change is accompanied by a configurational change of coordination from trans for 1 and 2 to cis for 3. The color change is associated with the superposition of ligand and Sm based electronic band energies and their intensities. Such white light emission by a single compound having contributions from different building components is quite rare.

Sensitized near infrared emission through supramolecular d → f energy transfer within an ionic Ru(ii)-Er(iii) pair.

The newly synthesized ionic triple salt Ru-Er, {[RuII(bpy)2(dbim)][ErIII(hfac)4][CF3COO]·H2O} (bpy = 2,2'-bipyridine; hfac- = hexafluoroacetylacetonate; dbim = 2,2'-dibenzimidazole) exhibits near-infrared (NIR) emission at 1535 nm by intermolecular Ru → Er (d → f) energy transfer across supramolecular interactions when pumped within the Ru(ii) 3MLCT band. It is the first such observation for a transition metal-lanthanide ionic pair.

[Inhibitory effect of lentivirus-mediated hTERTp-TK combined with hTERTp-tumstatin on human hepatocarcinoma HepG2 cells].

OBJECTIVE: To observe targeted expression of recombinant lentivirus-mediated (Lv)-hTERTp-TK and Lv-hTERTp-tumstatin in HepG2 cells, and explore the inhibitory effect of their combination on HepG2 cells both in vitro and in vivo. METHODS: Lv-hTERTp-TK and Lv-hTERTptumstatin were used to infect HepG2 and L02 cells at different MOIs. Transfection efficiency was observed by fluorescence microscopy. Expression of TK and tumstatin mRNA was detected by reverse-transcriptase PCR. Proliferation and apoptosis were detected by MTT and flow cytometry, respectively. The HepG2 cells were examined by real time-PCR and western blotting to determine expression level of bcl-2 and VEGF mRNA and protein.A murine hepatocellular carcinoma model was established by injecting 1 * 10(7) HepG2 cells into 30 BALB/c nude mice. The modeled mice were randomly divided into a control group, mock group, Lv-hTERTp-tumstatin group, Lv-hTERTp-TK group, and combination group for four weeks of injections at regular intervals of PBS, Lv-hTERTp-null, Lv-hTERTp-tumstatin, Lv-hTERTp-TK, and Lv-hTERTp-tumstatin plus Lv-hTERTp-TK, respectively.Changes in tumor volume and weight, and cell morphology of tumor and major organs, were assessed by hematoxylin-eosin staining.Microvascular density of tumor tissue and cell apoptosis were assessed by immunohistochemical and TUNEL staining, respectively. RESULTS: The Lv-infected HepG2 cells, and not the Lv-infected L02 cells, expressed TK and tumstatin. Lv-hTERTp-TK and Lv-hTERTp-tumstatin, alone or in combination, inhibited proliferation and increased apoptosis of the HepG2 cells, but the combination was more effective than either alone (P less than 0.05). None of the treatments affected proliferation or apoptosis of the L02 cells (P more than 0.05). The combination also led to a greater reduction of bcl-2 and VEGF than either alone (all, P less than 0.05). Tumor growth was significantly inhibited by the combination (P less than 0.05). In vivo, the combination treatment induced the greatest amount of apoptosis of the HepG2 cells. Cell morphology of major organs such as liver, spleen and kidney were similar to the control group. The combination also produced the most significant effect on tumor microvascular density (P less than 0.05) and the highest apoptosis index (P less than 0.05). CONCLUSION: The HTERT promoter can induce targeted expression of TK and tumstatin in HepG2 cells. Lv-hTERTp-TK combined with Lv-hTERTp-tumstatin can significantly inhibit proliferation and induce apoptosis of human HepG2 cells in vitro and in vivo, which may be related to down-regulation ofbcl-2 and VEGF.

Use of zona pellucida-bound sperm for intracytoplasmic sperm injection produces higher embryo quality and implantation than conventional intracytoplasmic sperm injection.

This goal of this study was to compare the outcomes of conventional intracytoplasmic sperm injection (ICSI; control group, n = 53 couples) and a modified ICSI technique using zona pellucida (ZP)-bound sperm for injection of oocytes (test group, n = 53 couples). The proportion of high-quality embryos (grades 1 and 2) and implantation rate were significantly higher in the test group than in the control group, but the difference in fetal heart pregnancy rate was not significant despite seven more pregnancies being obtained in the test group (26 pregnancies) versus the control group (19 pregnancies) following fresh embryo transfers.

[Drug-eluting stent implantation for unprotected left main coronary artery disease: immediate and long-term outcomes].

OBJECTIVE: To evaluate the safety and efficacy of drug-eluting stent (DES) implantation in selective patients with left main coronary artery disease. METHOD: From October 2002 to November 2007, 44 consecutive patients underwent percutaneous coronary interventions (PCI) on left main coronary artery lesions, including 5 patients with concurrent left ventricular dysfunction (ejection fraction<40%), 2 with chronic respiratory dysfunction and 5 with chronic renal failure. The findings in coronary angiography, procedural success rate, severe complications and the follow-up results of the patients were analyzed. RESULTS: The immediate procedural success rate was 100% in these patients without any severe complications. No non-fatal acute myocardial infarction or emergency coronary artery bypass grafting (CABG) was performed and death occurred in none of the cases during hospitalization. In the follow-up period for 14.2-/+9.3 (6-65) months after PCI, no subacute or late thromboses were found. One patient died from heart failure 4 months after PCI, and 6 patients (13.6%) experienced recurrent angina. Thirty-seven patients (84.1%) were free of any major cardiovascular events (MACE) after the procedure. A repeat coronary angiography was performed in 35 patients (79.5%) within 6 months after PCI, and 3 (8.6%) of them were confirmed to have restenosis, including 1 patient with distal bifurcation restenosis who were subsequently treated with CABG and two patients with side-branch ostium restenosis managed with cutting balloon dilation. CONCLUSIONS: Implantation of drug-eluting stents is safe and effective for management of left main coronary artery disease with good immediate and long-term outcomes.

[Asialofetuin-hTERT-TK/GCV targeted gene therapy and its bystander effect on HepG2].

OBJECTIVE: To observe the targeted therapeutic effects of plasmid AF-pGL3-hTERT-TK on HepG2 cells. METHODS: HepG2 cells were cultured and pGL3-hTERT-TK and AF-liposome were constructed. HepG2 and L02 cells were transfected with AF-pGL3-hTERT-TK. The growth, apoptosis of the cells and the bystander effects were studied using liquid scintillation analysis and tunnel and flow cytometry. RESULTS: After the suicide gene was inserted into the downstream of hTERT, TK was effectively driven by the hTERT promoter, making the TK highly expressed in the HepG2 cells. The AF made the therapeutic gene enter the HepG2 cells more easily by recognizing and combining the ASGPR receptor protein on the HepG2 cell surfaces and induced their apoptosis and suicide with bystander effect. The apoptosis rate was 85%+/-3% in the HepG2 cells whereas in the normal L02 hepatic cells it was 16%+/-2%. CONCLUSION: AF-pGL3-hTERT-TK can target and attack HepG2 cells and has almost no influence on normal L02 hepatic cells. AF-pGL3-hTERT-TK has a potential in the treatment of hepatocellular carcinomas.

[Effect of puerarin on ADMA-DDAH system in human umbilical vein endothelial cells cultured with oxidized free radical].

OBJECTIVE: To observe the effects of puerarin on activity of dimethylarginine dimethylaminohydrolase (DDAH) in human umbilical vein endothelial cells (HUVECs) cultured with oxidized free radical (OFR), to explore the effect of puerarin on metabolic mechanism of asymmetric dimethylarginine (ADMA). METHODS: HUVECs of the 3rd - 6th passage cultured with modified Jaffe's method were divided into 4 groups, the blank control group cultured with DMEM medium, the OFR group cultured with DMEM medium containing 0.1 mmol of OFR per liter, the puerarin group 1 and 2 cultured with DMEM medium containing 0.1 mmol of OFR per liter as well as 0.5 mg/ml and 1.0 mg/ml of puerarin respectively. After being incubated for 24 h, activity of nitric oxide synthase (NOS), contents of nitric oxide (NO), ADMA, endothelin (ET), and L-citrulline (L-cit) in the supernate were measured, and DDAH protein expression in the lysate was detected by Western blotting. RESULTS: Compared with those in the blank control group, ADMA and ET contents were higher, while the levels of NO and L-cit and the activity of NOS were lower markedly, but the DDAH expression changed insignificantly in the OFR group. These abnormalities were restored significantly in the puerarin groups. CONCLUSION: The increase of ADMA in OFR injured HUVECs was correlated with the reduction of DDAH activity and irrelevant to DDAH expression. Puerarin could promote ADMA metabolism through increasing DDAH activity, and improve NOS activity, thus to reduce the impairing of OFR on endothelial function.