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The Hulunbuir region, known for its diverse terrain and rich wildlife, is a hotspot for various natural epidemic diseases. Between 2021 and 2023, we collected 885 wild rodent samples from this area, representing three families, seven genera, and eleven species. Metagenomic analysis identified three complete nucleic acid sequences from the S, M, and L segments of the Hantaviridae family, which were closely related to the Khabarovsk virus. The nucleotide coding sequences for S, M, and L (1392 nt, 3465 nt, and 6491 nt, respectively) exhibited similarities of 82.34%, 81.68%, and 81.94% to known sequences, respectively, while protein-level analysis indicated higher similarities of 94.92%, 94.41%, and 95.87%, respectively. Phylogenetic analysis placed these sequences within the same clade as the Khabarovsk, Puumala, Muju, Hokkaido, Topografov, and Tatenalense viruses, all of which are known to cause febrile diseases in humans. Immunofluorescence detection of nucleic acid-positive rodent kidney samples using sera from patients with hemorrhagic fever and renal syndrome confirmed the presence of viral particles. Based on these findings, we propose that this virus represents a new member of the Hantaviridae family, tentatively named the Amugulang virus, after its primary distribution area.
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Background: Breast cancer ranks as one of the most prevalent malignancies among women globally, with increasing incidence rates. Physical activity, particularly exercise, has emerged as a potentially significant modifier of cancer prognosis, influencing tumor biology and patient outcomes. Methods: Using a murine breast cancer model, we established a control and an exercise group, where the latter was subjected to 21 days of voluntary running. RNA Sequencing, bioinformatics analysis, pan-cancer analysis, and cell experiments were performed to validate the underlying mechanisms. Results: We observed that exercise significantly reduced tumor size and weight, without notable changes in body weight, suggesting that physical activity can modulate tumor dynamics. mRNA sequencing post-exercise revealed substantial downregulation of CD300E in the exercise group, accompanied by alterations in critical pathways such as MicroRNAs in cancers and the Calcium signaling pathway. Expanding our analysis to a broader cancer spectrum, CD300E demonstrated significant expression variability across multiple cancer types, with pronounced upregulation in myeloma, ovarian, lung, and colorectal cancers. This upregulation was correlated with poorer prognostic outcomes, emphasizing CD300E's potential role as a prognostic marker and therapeutic target. Moreover, CD300E expression was associated with cancer cell proliferation and apoptosis. Conclusion: The study highlights the dual role of exercise in modulating gene expression relevant to tumor growth and the potential of CD300E as a target in cancer therapeutics. Further research is encouraged to explore the mechanisms by which exercise and CD300E influence cancer progression and to develop targeted strategies that could enhance patient outcomes in clinical settings.
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Regulación Neoplásica de la Expresión Génica , Animales , Femenino , Humanos , Ratones , Apoptosis/genética , Biomarcadores de Tumor/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Neoplasias de la Mama/terapia , Línea Celular Tumoral , Proliferación Celular , Regulación hacia Abajo , Neoplasias/genética , Condicionamiento Físico Animal , Pronóstico , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismoRESUMEN
Background: Breast cancer, one of the most prevalent malignancies among women worldwide, has rising incidence rates. Physical activity, particularly exercise, has emerged as a significant modifier of cancer prognosis, influencing both tumor biology and patient outcomes. Methods: In this study, we utilized a murine breast cancer model, dividing mice into a control group and an exercise group; the latter underwent 21 days of voluntary running. We conducted RNA sequencing, bioinformatics analysis, pan-cancer analysis, and cellular experiments to investigate the underlying mechanisms influenced by exercise. Results: Exercise led to a significant reduction in tumor size and weight. Post-exercise mRNA sequencing indicated a notable upregulation of THSD7B in the exercised mice, with significant alterations observed in pathways such as MicroRNAs in cancers and the Calcium signaling pathway. In a broader cancer context, THSD7B showed considerable expression variability, being significantly downregulated in several cancers, correlating with positive prognostic outcomes in PRAD, LAML, KIRC, and GBM and highlighting its potential role as a prognostic marker and therapeutic target. THSD7B expression was also negatively associated with processes of breast cancer cell proliferation, migration, and invasion. Conclusion: This study underscores the dual role of exercise in modulating gene expression relevant to tumor growth and highlights the potential of THSD7B as a therapeutic target in cancer. Future research should further explore the specific mechanisms by which exercise and THSD7B influence cancer progression and develop immunotherapy-enhanced strategies to change patient outcomes in clinical settings.
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Condicionamiento Físico Animal , Animales , Ratones , Femenino , Pronóstico , Humanos , Regulación Neoplásica de la Expresión Génica , Neoplasias de la Mama/genética , Neoplasias de la Mama/terapia , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/patología , Proliferación Celular , Línea Celular Tumoral , Biomarcadores de Tumor/genéticaRESUMEN
Introduction: Ticks are the most important obligate blood-feeding vectors of human pathogens. With the advance of high-throughput sequencing, more and more bacterial community and virome in tick has been reported, which seems to pose a great threat to people. Methods: A total of 14 skin specimens collected from tick-bite patients with mild to severe symptoms were analyzed through meta-transcriptomic sequencings. Results: Four bacteria genera were both detected in the skins and ticks, including Pseudomonas, Acinetobacter, Corynebacterium and Propionibacterium, and three tick-associated viruses, Jingmen tick virus (JMTV), Bole tick virus 4 (BLTV4) and Deer tick mononegavirales-like virus (DTMV) were identified in the skin samples. Except of known pathogens such as pathogenic rickettsia, Coxiella burnetii and JMTV, we suggest Roseomonas cervicalis and BLTV4 as potential new agents amplified in the skins and then disseminated into the blood. As early as 1 day after a tick-bite, these pathogens can transmit to skins and at most four ones can co-infect in skins. Discussion: Advances in sequencing technologies have revealed that the diversity of tick microbiome and virome goes far beyond our previous understanding. This report not only identifies three new potential pathogens in humans but also shows that the skin barrier is vital in preventing horizontal transmissions of tick-associated bacteria or virus communities to the host. It is the first research on patients' skin infectome after a tick bite and demonstrates that more attention should be paid to the cutaneous response to prevent tick-borne illness.
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Coxiella burnetii , Rickettsia , Mordeduras de Garrapatas , Enfermedades por Picaduras de Garrapatas , Garrapatas , Virus , Animales , Humanos , Garrapatas/microbiología , Piel , Virus/genéticaRESUMEN
The increasing prevalence and expanding distribution of tick-borne viruses globally have raised health concerns, but the full repertoire of the tick virome has not been assessed. We sequenced the meta-transcriptomes of 31 different tick species in the Ixodidae and Argasidae families from across mainland China, and identified 724 RNA viruses with distinctive virome compositions among genera. A total of 1,801 assembled and complete or nearly complete viral genomes revealed an extensive diversity of genome architectures of tick-associated viruses, highlighting ticks as a reservoir of RNA viruses. We examined the phylogenies of different virus families to investigate virome evolution and found that the most diverse tick-associated viruses are positive-strand RNA virus families that demonstrate more ancient divergence than other arboviruses. Tick-specific viruses are often associated with only a few tick species, whereas virus clades that can infect vertebrates are found in a wider range of tick species. We hypothesize that tick viruses can exhibit both 'specialist' and 'generalist' evolutionary trends. We hope that our virome dataset will enable much-needed research on vertebrate-pathogenic tick-associated viruses.
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Virus ARN , Garrapatas , Virus , Animales , Virus ARN/genética , Genoma Viral/genética , ARNRESUMEN
Background: Jingmen tick virus (JMTV) has attracted great attention due to its potential pathogenicity in humans and its transmission by ticks. Dermacentor silvarum (D. silvarum) is one of the dominant tick species in northeastern China, and can transmit many pathogens to humans and animals. However, there have been no report of transmission of JMTV by D. silvarum. Materials and Methods: Ticks were collected from vegetation at the Aershan Port in Inner Mongolia in April 2019. And we do attempt to infect D. silvarum with JMTV by the immersion technique in laboratory conditions. The transmission of JMTV was examined by reverse transcriptase PCR, fluorescence in situ hybridization, and indirect immunofluorescence assay. Statistical analysis was performed using SPSS 24.0. Results: We found that JMTV may only be maintained in the tick without replication, and could not be transmitted to a host following transstadial transmission. Moreover, no virus colonization was found in the midgut or salivary glands of unfed D. silvarum; therefore, D. silvarum may not be susceptible to JMTV infection and therefore unlikely to carry and transmit JMTV. Conclusion: Our study has to some extent filled the knowledge gap regarding the possibility of JMTV transmission by a medically important tick vector, D. silvarum.
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Dermacentor , Animales , China/epidemiología , Dermacentor/genética , Vectores de Enfermedades , Humanos , Hibridación Fluorescente in Situ/veterinariaRESUMEN
PURPOSE: The association between the process of postoperative pneumonia and lung cancer recurrence remains elusive in lung cancer surgery. Herein, the association between postoperative pneumonia and lung cancer recurrence was investigated, emphasizing the warning role of postoperative specific pneumonia in primary lung cancer resection patients. METHODS: The occurrence of postoperative pneumonia was assessed in 4-6 months (PPFS), 7-12 months (PPST), and lung cancer recurrence within 1 year (LRO) in 332 patients. The primary outcome was the development of PPST and LRO according to PPFS occurrence. The relevant risk factors of PPFS, PPST, and LRO were identified through multivariable regression analysis. RESULTS: During follow-up, 151 (45.48%) participants experienced PPFS. Irrespective of the existing postoperative pneumonia in 1-3 months (PPOT), PPFS significantly increased the risk of PPST (P < 0.01) and LRO (P < 0.01), and persistent PPST further increased the risk of LRO (P < 0.001). The generalized estimating equation identified chemotherapy as an independent risk factor for PPFS and PPST. CONCLUSION: PPFS was associated with the increased risk of PPST and LRO. Postoperative pulmonary inflammation assessed 4 months post-surgery also significantly influenced LRO development, indicating a need for close follow-up of lung inflammatory conditions to improve patient outcomes.
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Among arthropod vectors, ticks transmit the most diverse human and animal pathogens, leading to an increasing number of new challenges worldwide. Here we sequenced and assembled high-quality genomes of six ixodid tick species and further resequenced 678 tick specimens to understand three key aspects of ticks: genetic diversity, population structure, and pathogen distribution. We explored the genetic basis common to ticks, including heme and hemoglobin digestion, iron metabolism, and reactive oxygen species, and unveiled for the first time that genetic structure and pathogen composition in different tick species are mainly shaped by ecological and geographic factors. We further identified species-specific determinants associated with different host ranges, life cycles, and distributions. The findings of this study are an invaluable resource for research and control of ticks and tick-borne diseases.
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Variación Genética/genética , Enfermedades por Picaduras de Garrapatas/microbiología , Garrapatas/genética , Animales , Línea Celular , Vectores de Enfermedades , Especificidad del Huésped/genéticaRESUMEN
BACKGROUND: Fructus Psoraleae is the seed of Psoralea corylifolia Linn. Fructus Psoraleae has been shown to be effective in treating some skin diseases, such as vitiligo. As a main ingredient in five types of herbs in the Qubaibabuqi tablet formula, Fructus Psoraleae plays an important role in the treatment of vitiligo. Fructus Psoraleae has potential hepatotoxicity, thus Qubaibabuqi tablets also have potential liver toxicity. CASE PRESENTATION: A 53-year-old woman who was diagnosed with vitiligo in September 2017 was treated with Qubaibabuqi tablets. After approximately 7 months of treatment, the patient developed a severe, diffuse yellow staining of the skin and sclera in March 2018. On admission, she was diagnosed with acute cholestatic hepatitis associated with Fructus Psoraleae. Despite receiving active treatment, her condition rapidly deteriorated and she died 5 days later due to acute liver failure and multiple organ dysfunction. To the best of our knowledge, there are only six reported cases of liver injury associated with Fructus Psoraleae described in the English language literature; however, cases of acute liver failure associated with the use of Fructus Psoraleae have not been described. CONCLUSION: As a main ingredient in the Qubaibabuqi tablet formula, Fructus Psoraleae has potential hepatotoxicity. This potentially fatal adverse effect should be considered when physicians prescribe Qubaibabuqi tablets.
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Medicamentos Herbarios Chinos/efectos adversos , Fallo Hepático Agudo/inducido químicamente , Psoralea , Adulto , Colecistitis Aguda/inducido químicamente , Medicamentos Herbarios Chinos/uso terapéutico , Resultado Fatal , Femenino , Humanos , Masculino , Persona de Mediana Edad , Vitíligo/tratamiento farmacológico , Adulto JovenRESUMEN
Aging leads to skeletal muscle atrophy (i.e., sarcopenia), and muscle fiber loss is a critical component of this process. The mechanisms underlying these age-related changes, however, remain unclear. We show here that mTORC1 signaling is activated in a subset of skeletal muscle fibers in aging mouse and human, colocalized with fiber damage. Activation of mTORC1 in TSC1 knockout mouse muscle fibers increases the content of morphologically abnormal mitochondria and causes progressive oxidative stress, fiber damage, and fiber loss over the lifespan. Transcriptomic profiling reveals that mTORC1's activation increases the expression of growth differentiation factors (GDF3, 5, and 15), and of genes involved in mitochondrial oxidative stress and catabolism. We show that increased GDF15 is sufficient to induce oxidative stress and catabolic changes, and that mTORC1 increases the expression of GDF15 via phosphorylation of STAT3. Inhibition of mTORC1 in aging mouse decreases the expression of GDFs and STAT3's phosphorylation in skeletal muscle, reducing oxidative stress and muscle fiber damage and loss. Thus, chronically increased mTORC1 activity contributes to age-related muscle atrophy, and GDF signaling is a proposed mechanism.
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Envejecimiento/metabolismo , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patología , Estrés Oxidativo , Animales , Células Cultivadas , Humanos , Diana Mecanicista del Complejo 1 de la Rapamicina/antagonistas & inhibidores , Ratones , Ratones Noqueados , Ratones Transgénicos , Proteína 1 del Complejo de la Esclerosis Tuberosa/deficiencia , Proteína 1 del Complejo de la Esclerosis Tuberosa/metabolismoRESUMEN
The complete mitochondrial genome of Amblyomma geoemydae is reported for the first time in this study. Its entire mitogenome is 14,780 bp in length, contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and two non-coding regions. The phylogenetic analysis by Maximum-likelihood method show that A. geoemydae and the others of genus Amblyomma are in the same clade, indicating that A. geoemydae belongs to the genus Amblyomma.
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Ustilaginoidea virens is an important fungus that causes rice false smut disease. This disease significantly reduces both grain yield and quality. Various methods have been developed for the detection of U. virens but most of these methods need sophisticated equipment such as a thermal cycler. Here, we present a loop-mediated isothermal amplification (LAMP) assay for the specific detection of U. virens. This assay used a specific region of the UvG-ß1 gene (212-bp region) to design six LAMP primers. The LAMP assay was optimized by the combination of rapidity, simplicity, and high sensitivity for the detection of about 1 pg of target genomic DNA in the reaction whereas, with polymerase chain reaction (PCR), there was no amplification of DNA with concentrations less than 1 ng. Among the genomic DNA of 22 fungus species and two strains of U. virens, only the tube containing the DNA of U. virens changed to yellowish green with SYBR Green I. The color change was indicative of DNA amplification. No DNA was amplified from either the other 22 fungus species or the negative control. Moreover, 20 spikelets and 22 rice seed samples were used for the detection of rice false smut via LAMP. The results were comparable with conventional PCR. We conclude that gene UvG-ß1 coupled with LAMP assay, can be used for the detection and identification of U. virens gene via LAMP.
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Hypocreales/genética , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Cartilla de ADN/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa , Semillas/microbiología , Factores de TiempoRESUMEN
α-MnO2 is a promising material for ozone catalytic decomposition and the oxygen vacancy is often regarded as the active site for ozone adsorption and decomposition. Here, α-MnO2 nanowire with tunable K+ concentration was prepared through a hydrothermal process in KOH solution. High concentration K+ in the tunnel can expand crystal cell and break the charge balance, leading to a lower average oxidation state (AOS) of Mn, which means abundant oxygen vacancy. DFT calculation has also proven that the samples with higher K+ concentration exhibit lower formation energy for oxygen vacancy. Due to the enormous active oxygen vacancies existing in the α-MnO2 nanowire, the lifetime of the catalyst (corresponding to 100% ozone removal rate, 25 °C) is increased from 3 to 15 h. The FT-IR results confirmed that the accumulation of intermediate oxygen species on the catalyst surface is the main reason why it is deactivated after long time reaction. In this work, the performance of the catalyst has been improved because the abundant active oxygen vacancies are fabricated by the electrostatic interaction between oxygen atoms inside the tunnels and the introduced K+, which offers us a new perspective to design a high efficiency catalyst and may promote manganese oxide for practical ozone elimination.
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Ozono , Catálisis , Oxidación-Reducción , Oxígeno , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
OBJECTIVE: To evaluate the application of small intestine double stoma and succus entericus reinfusion in the patients with severe intra-abdominal infection. METHODS: Ten patients with high intestinal perforation from February 2005 to November 2014 were enrolled in the study. All the cases received emergency operation. Small bowel with intestinal perforation was resected, and double stoma was applied in the proximal and distal small intestine. When abdominal infection under control, total enteral nutrition was successfully administered from nasogastric tube. The succus entericus from the proximal intestine was collected and transfused back to the distal intestine. Stool was collected and fecal nitrogen, fat and carbohydrate contents were determined. Related serum protein levels were measured. RESULTS: As compared to pre-reinfusion, the absorption rate of carbohydrate [(90.9±7.8)% vs. (82.7±15.2)%], fat [(87.6±6.4)% vs. (59.1±10.8)%], and nitrogen [(82.4±9.8)% vs. (67.2±15.4)%] increased after succus entericus reinfusion (P<0.05). The serum protein levels increased significantly as well[fibronectin: (285.6±3.6) vs. (157.0±22.6) mg/L, P<0.01; transferrin: (4.86±0.21) vs. (3.60±0.25) g/L, P<0.05; pre-albumin: (291.3±112.5) vs. (199.1±53.3) mg/L, P<0.05]. CONCLUSION: Small intestine double stoma and succus entericus reinfusion are effective in improving the absorption of carbohydrate, fat and nitrogen in the patients with severe intra-abdominal infection.
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Intestino Delgado , Infecciones Intraabdominales , Nutrición Enteral , Humanos , Perforación Intestinal , Secreciones Intestinales , Estomas QuirúrgicosRESUMEN
OBJECTIVE: The objective was to investigate whether the combination of simvastatin and aspirin treatment prolongs the survival time of the heart allograft in rat and its underlying mechanism. METHODS: Heterotopic heart transplantation was performed using Wistar rats as donors and Sprague-Dawley (SD) rats as recipients. The SD rats were randomly divided into five groups (n=20/group): sham, HT (heart transplantation), HT+simvastatin (HT+S), HT+aspirin (HT+A), and HT+aspirin+simvastatin (HT+A+S). After transplantation, at 3, 7, 10, 15, 20, 30, and 40 days, the endothelial nitric oxide synthase (eNOS) expression was assessed by immunohistological staining; nitric oxide (NO) levels were analyzed by Griess assay; the activation of CD4(+)CD25(+) T regulatory lymphocytes (Tregs) was analyzed by flow cytometry; and pathological changes in the graft heart were determined by histology. RESULTS: Combined treatment of hearts with simvastatin and aspirin significantly prolonged the mean survival time of heart allografts [8 ± 1.2 days (n=18), 20 ± 3.4 days (n=19), 21 ± 2.8 days (n=19), and 39 ± 5.3 days (n=19) for HT, HT+S, HT+A, and HT+A+S group, respectively; HT vs. HT+A+S, P<.001; HT vs. HT+S or HT+A, P<.05]. In addition, CONCLUSIONS: Simvastatin given in combination with aspirin delayed the development of pathological changes in the myocardium, reduced vascular damage and prolonged the survival time of cardiac allograft. The underlying mechanism is linked with CD4(+)CD25(+)-Treg-cell-induced immune tolerance and enhanced vascular endothelial cell protection.
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Aspirina/administración & dosificación , Células Endoteliales/efectos de los fármacos , Supervivencia de Injerto/efectos de los fármacos , Trasplante de Corazón/métodos , Simvastatina/administración & dosificación , Linfocitos T Reguladores/inmunología , Aloinjertos/inmunología , Animales , Modelos Animales de Enfermedad , Citometría de Flujo , Rechazo de Injerto/inmunología , Supervivencia de Injerto/inmunología , Inmunohistoquímica , Ratas , Ratas Sprague-Dawley , Ratas WistarRESUMEN
OBJECTIVES: To investigate expression of pentraxin 3, long (PTX3) in patients with acute coronary syndrome (ACS) and its correlation with matrix metalloproteinase-9 (MMP-9) and C-reactive protein (CRP) levels. METHODS: Patients with ACS were randomly assigned to the ACS group (subdivided into unstable angina pectoris [UAP] and acute myocardial infarction [AMI]). Healthy participants and patients with stable angina pectoris (SAP) were enrolled as controls. Mononuclear cell PTX3 expression, and serum MMP-9 and CRP levels, were measured by enzyme-linked immunosorbent assay. RESULTS: The ACS group comprised 200 patients (80 in the UAP subgroup; 120 in the AMI subgroup). The control group comprised 130 participants (80 healthy volunteers and 50 patients with SAP). PTX3 expression was significantly higher in the ACS group compared with controls (3.64 ± 0.49 versus 1.85 ± 0.65 ng/ml), and significantly higher in the AMI compared with the UAP subgroup (5.44 ± 0.47 versus 3.39 ± 0.59 ng/ml). Serum MMP-9 and CRP levels were significantly higher in the ACS group compared with controls (48.55 ± 14.22 versus 23.14 ± 0.62 ng/ml; 4.88 ± 1.76 versus 1.26 ± 0.19 ng/ml, respectively), and significantly higher in the AMI compared with the UAP subgroup (58.13 ± 7.24 versus 31.77 ± 3.61 ng/ml; 5.80 ± 1.46 versus 3.27 ± 0.83 ng/ml, respectively). PTX3 expression, and MMP-9 and CRP levels in the SAP subgroup, were not significantly different from the healthy participants. PTX3 expression positively correlated with MMP-9 and CRP levels. CONCLUSIONS: In patients with ACS, peripheral blood mononuclear cell PTX3 expression, and serum MMP-9 and CRP levels, were significantly enhanced compared with controls; in addition, PTX3 expression positively correlated with MMP-9 and CRP levels. PTX3 may be involved in ACS pathogenesis.
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Síndrome Coronario Agudo/genética , Angina Estable/genética , Angina Inestable/genética , Proteína C-Reactiva/genética , Metaloproteinasa 9 de la Matriz/genética , Componente Amiloide P Sérico/genética , Síndrome Coronario Agudo/sangre , Síndrome Coronario Agudo/diagnóstico , Síndrome Coronario Agudo/patología , Anciano , Angina Estable/sangre , Angina Estable/diagnóstico , Angina Estable/patología , Angina Inestable/sangre , Angina Inestable/diagnóstico , Angina Inestable/patología , Proteína C-Reactiva/metabolismo , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Femenino , Regulación de la Expresión Génica , Humanos , Leucocitos Mononucleares/metabolismo , Leucocitos Mononucleares/patología , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Persona de Mediana Edad , Pronóstico , Componente Amiloide P Sérico/metabolismoRESUMEN
BACKGROUND: The spleen is an active lymphoid organ. The effect of splenectomy on the immune response remains unclear. This study investigated whether splenectomy can induce immune tolerance and has a beneficial role in cardiac allograft. METHODS: Wistar rats were used for heart donors. The Sprague-Dawley (SD) rats designated as the recipients of heart transplantation (HT) were randomly assigned into four groups: sham, splenectomy, HT, splenectomy + HT. The survival of transplanted hearts was assessed by daily checking of abdominal palpation. At various time points after transplantation, the transplanted hearts were collected and histologically examined; the level of CD4+CD25+ T regulatory lymphocytes (Tregs) and rate of lymphocyte apoptosis (annexin-v+ PI+ cells) in the blood were analyzed by using flow cytometric method. RESULTS: 1) Splenectomy significantly prolonged the mean survival time of heart allografts (7 ± 1.1 days and 27 ± 1.5 days for HT and splenectomy + HT, respectively; n = 12-14/group, HT vs. splenectomy + HT, p < 0.001); 2) Splenectomy delayed pathological changes (inflammatory cell infiltration, myocardial damage) of the transplanted hearts in splenectomy + HT rats; 3) The level of CD4+CD25+ Tregs in the blood of splenectomized rats was significantly increased within 7 days (2.4 ± 0.5%, 4.9 ± 1.3% and 5.3 ± 1.0% for sham, splenectomy and splenectomy + HT, respectively; n = 15/group, sham vs. splenectomy or splenectomy + HT, p < 0.05) after splenectomy surgery and gradually decreased to baseline level; 4) Splenectomy increased the rate of lymphocyte apoptosis (day 7: 0.3 ± 0.05%, 3.9 ± 0.9% and 4.1 ± 0.9% for sham, splenectomy and splenectomy + HT, respectively; n = 15/group, sham vs. splenectomy or splenectomy + HT, p < 0.05) in a pattern similar to the change of the CD4+CD25+ Tregs in the blood. CONCLUSIONS: Splenectomy inhibits the development of pathology and prolongs the survival time of cardiac allograft. The responsible mechanism is associated with induction of immune tolerance via elevating CD4+CD25+ Tregs and increasing lymphocyte apoptosis.
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Supervivencia de Injerto/inmunología , Trasplante de Corazón , Tolerancia Inmunológica , Inmunidad Celular , Esplenectomía , Linfocitos T Reguladores/inmunología , Aloinjertos , Animales , Modelos Animales de Enfermedad , Ratas , Ratas Sprague-Dawley , Ratas WistarRESUMEN
OBJECTIVE: To investigate the expression of kir2.1 protein in primary cultured sinus node cells and establish a reliable technique to locate, culture and characterize neonatal rat sinus node cells. METHODS: In paraffin sections, the location and morphology of the neonatal rat sinus node cells were observed by HE staining, silver nitrate staining, myelin staining and phosphotungstic acid-hematoxylin (PTAH) staining. Primary cell culture from the neonatal rat sinus node was conducted to observe the spontaneous contraction frequency, cell morphology and kir2.1 protein expression. RESULTS: Combination of the 3 staining methods allowed accurate localization of the sino-atrial nodal (SAN) tissue, and among the cultured cells in the SAN, at least 3 distinct types of cells with spontaneous contraction were observed. The majority of the contracting cells were spindle cells and their construction and impulse frequency indicated their identity as pacemaker cells, while the triangular and irregular cells resembled the atrial muscle cells. A lower expression level of kir2.1 protein was detected in SAN cells than in the atrial and ventricular myocytes of the neonatal rats. CONCLUSION: Combination of silver nitrate staining, myelin staining and PTAH staining identifies the exact location of the sinus node tissue, and cultured sinus node cells have lower expression of kir2.1 protein than the atrial and ventricular myocytes of neonatal rats.
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Técnicas de Cultivo de Célula/métodos , Canales de Potasio de Rectificación Interna/metabolismo , Nodo Sinoatrial/citología , Animales , Células Cultivadas , Ratas , Ratas Sprague-Dawley , Nodo Sinoatrial/metabolismoRESUMEN
BACKGROUND: Most studies about FHIT protein expression were performed in normal tracheal epithelium, precancerous lesions and lung cancer tissues respectively, but not in the course of malignant transformation of lung cancer. The aim of this study is to detect the changes of FHIT protein expression during malignant transformation of immortalized human bronchial epithelial cells (BEAS-2B) induced by tobacco-specific nitrosamine (NNK), and to explore its significance. METHODS: BEAS-2B cells were induced to malignantly transform (BEAS-2B NNK) by 500mg/L NNK, and FHIT protein expression was detected in the different passages of BEAS-2B NNK and BEAS-2B cells by SP immunocytochemistry. RESULTS: Part 1: Model of malignant transformation of BEAS-2B cells induced by NNK was established. (1) The serum resistance was significantly increased in the 5th passage of BEAS-2B NNK cells. (2) The anchorage independent growth (soft agar colony formation) appeared in the 15th passage of BEAS-2B NNK cells. (3) The ultrastructure of the 20th passage of BEAS-2B NNK cells showed obvious heteromorphy characterization. (4) The 25th passage of BEAS-2B NNK cells developed into tumors in nude mice, which were well differentiated squamous cell carcinoma. Part 2: FHIT protein was steadily expressed in the different passages of BEAS-2B cells (P > 0.05). FHIT protein expression was obviously decreased from 5th to 15th passage of BEAS-2B NNK cells, but it was unexpectedly overexpressed in the 25th passage. CONCLUSIONS: (1) The model of malignant transformation of BEAS-2B cells induced by NNK (500mg/L) can be established successfully and may be used for investigation of molecular biological mechanisms of lung cancer, especially for smoking-related cases. (2) Decrease of FHIT protein expression might be an early event, however, its overexpression in the late passages should be further studied.