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1.
Sci Total Environ ; 923: 171541, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38453084

RESUMO

Asian over-land aerosols are complexities due to a mixture of anthropogenic air pollutants and natural dust. The accuracy of the aerosol optical thickness (AOT) retrieved from the satellite is crucial to their application in the aerosol data assimilation system. Fusion of AOTs with high spatiotemporal resolution from next-generation geostationary satellites such as Fengyun-4B (FY-4B) and Himawari-9, provides a new high-quality dataset capturing the aerosol spatiotemporal variability for data assimilation. This study develops a complete fusion algorithm to estimate the optimal AOT over-land in Asia from September 2022 to August 2023 at 10 km × 10 km resolution with high efficiency. The data fusion involves four steps: (1) investigating the spatiotemporal variability of FY-4B AOT within the past 1 h and 12 km radius calculation domain; (2) utilizing the aerosol spatiotemporal variability characteristics to estimate FY-4B pure and hourly merged AOTs; (3) performing bias corrections for FY-4B and Himwari-9 hourly merged AOT for different observation times and seasons considering pixel-level errors for each satellite; (4) fusing the bias-corrected FY-4B and Himawari-9 hourly merged AOT based on maximum-likelihood estimation (MLE) method. Compared to the original FY-4B AOT, validation with AERONET observation confirms that the root mean square error (RMSE) of hourly merged FY-4B AOT decreases by around 40.6 % and the correlation coefficient (CORR) increases by about 27.8 %. Compared to FY-4B and Himawari-9 merged AOT, the fused AOT significantly decreases (increases) RMSE (CORR) by around 24.7 % (7.3 %) and 20.2 % (5.6 %). In addition, fused AOT is double the number of single-sensor merged AOT. Fusion aerosol map accurately describes the spatial and temporal variations in Asian regions controlled by air pollution and dust storms. Further studies are required for other landscapes with different satellite combinations to promote the application in the data assimilation system.

3.
Ann Transl Med ; 10(2): 38, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35282082

RESUMO

Background: Our research team has identified a biological active component, panaxadiol saponins component (PDS-C) isolated from total saponins of panax ginseng as a potential targeted drug for treating hemocytopenia. PDS-C possesses dual activities, namely that of promoting hematopoiesis and regulating immune function. Our study is to observe effects of PDS-C on promoting hematopoiesis in normal and aplastic anemia (AA) mice, furthermore, to explore its possible mechanism. Methods: Bone marrow nucleated cells were cultured for colony forming assay of CFU-GM, CFU-E and CFU-MK in the presence of PDS-C at different concentration. The proliferation and differentiation-related genes expression profile was analyzed with DNA membrane microarray. The mRNA expression levels and protein phosphorylated state of GATA-1, GATA-2 transcription factors and AKT-1, MAPK14 protein kinases were detected by RT-qPCR and Western blot, the DNA binding activity and components of GATA-DNA complex were analyzed by EMSA and antibody gel supershift assay. Results: In response to PDS-C at 10, 25 and 50 mg/L, the bone marrow colony numbers of CFU-GM, CFU-E and CFU-MK increased significantly by 25.7%±3.1% to 42.4%±4.5% respectively in normal mice, and 29.7%±3.7% to 53.2%±7.1% in AA mice. The gene microarray profile initiated by PDS-C provided the up-regulated genes by more than 3 times, which can be classified into 11 categories according to their functions, including GATA-1, GATA-2, and AKT-1, MAPK14. The mRNA expression levels of GATA-1, GATA-2 were consistent with their gene microarray profile in PDS-C treated erythroid and megakaryocytic hematopoietic cells. Meanwhile, PDS-C could not only up-regulate expression levels of GATA-1, GATA-2 proteins, but also enhance phosphorylated activity state. Furthermore, PDS-C obviously enhanced binding activity of GATA protein with DNA in erythroid and megakaryocytic cells, and the main composition of GATA-DNA complex was GATA-2 and GATA-1. Conclusions: PDS-C displays the role to promote proliferation and induce differentiation for hematopoietic cells. Its action mechanism may involve in GATA-1, GATA-2 transcription factors, including up-regulating mRNA and protein expression, enhancing DNA binding activity, phosphorylated functional activity and up-regulating AKT-1, MAPK14 protein kinases as the upstream signaling molecule for activation GATA-1, GATA-2 respectively in hematopoietic cells.

4.
Theor Appl Climatol ; 147(3-4): 1675-1684, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35095143

RESUMO

The deep blue sky is an indicator of a lower concentration of aerosols and a cloudless sky. With increasing human emissions, a trend towards days with fewer deep blue skies might indicate a decline in a good living environment for humans. This study investigates the long-term changes of the deep blue sky in China from 1980 to 2018. Due to a lack of direct measurements, we use atmospheric visibility and low cloud cover to classify blue sky days into three grades: light blue day, medium blue day, and deep blue day. Climatologically, annual deep blue days increase from southeast China to northwest China, with the maximum number in Xinjiang and eastern Inner Mongolia and the minimum number in western Qinghai and southern Hebei. From 1980 to 2018, annual deep blue days show a prominent decreasing trend in most of China, with area-mean annual deep blue days decreasing by -0.48 days per year (d/y) in China, and the variation becomes more obvious after 2013. The maximum decreasing trend is observed in eastern China. The most prominent decreases of deep blue days are seen in winter. Both air pollution and the change in meteorological conditions contribute to the decrease of wintertime deep blue days in China. Specifically, the decrease in surface wind speed hinders the cleaning of air by winds, the increase in surface air temperature, and decrease in relative humidity is favorable for low cloud increase, and the increasing emission of pollution reduces atmospheric visibility. Supplementary Information: The online version contains supplementary material available at 10.1007/s00704-021-03898-1.

5.
Chin J Integr Med ; 27(2): 131-136, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32418174

RESUMO

OBJECTIVE: To explore the clinical effect and adverse reactions of Strychnos nux-vomica in bortezomib-induced peripheral neuropathy (BIPN) of patients with multiple myeloma (MM). METHODS: A total of 19 MM patients with BIPN were enrolled and Nux Vomica Capsule (NVC, 0.4 g, thrice daily) were orally administrated for 30 days. Comparative analysis on parameters between pre- and post-therapy, including peripheral neuropathy (PN) grade, neurotoxicity score, Chinese medicine (CM) syndrome score, total neuropathy score (TNS), coagulation function, and serum nerve growth factor (NGF) levels were conducted. The adverse events were monitored. RESULTS: In BIPN of MM patients who received NVC, PN grade was lowered, neurotoxicity score was obviously decreased (P⩽0.01), and both CM syndrome score and TNS were remarkably decreased (P<0.01). After the therapy, activated partial thromboplastin time was prolonged (P<0.01) and fibrinogen was declined (P<0.05), showing improvement in the hypercoagulable state of patients. No significant difference of NGF recovery degrees was detected between pre- and post-therapy (P>0.05). No evident adverse reactions were observed during the course of treatment. CONCLUSION: Strychnos nux-vomica L. has significantly effect with a good safety in treatment of BIPN in MM patients.


Assuntos
Mieloma Múltiplo , Doenças do Sistema Nervoso Periférico , Strychnos nux-vomica , Bortezomib/efeitos adversos , Humanos , Mieloma Múltiplo/tratamento farmacológico , Doenças do Sistema Nervoso Periférico/induzido quimicamente , Doenças do Sistema Nervoso Periférico/tratamento farmacológico , Sementes
6.
Sci Total Environ ; 763: 143003, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33168256

RESUMO

In this study, simulations of the annual mean aerosol budget, aerosol optical properties, and surface mass concentration in 2006 in China are performed with two aerosol interactive global atmosphere models, namely, the Nonhydrostatic ICosahedral Atmospheric Model (NICAM) coupled with the Spectral Radiation Transport Model for Aerosol Species (SPRINTARS) and the Beijing Climate Center Atmospheric General Circulation Model (BCC_AGCM) coupled with the Canadian Aerosol Module (CAM) online. The observed and simulated aerosol optical depths (AODs) exhibit similar horizontal distributions with large values over eastern and central China, and sulfate aerosols contribute the main differences between the AODs simulated by NICAM and BCC_AGCM. The simulated sulfate and dust surface concentrations are more consistent with observations compared with the simulated carbonaceous surface concentrations, and both models can reproduce the decreasing tendency of the sulfate surface concentration from urban sites to rural sites. However, the dust emission and deposition levels in China simulated by BCC_AGCM are three times as high as those simulated by NICAM, and the major sink processes of the anthropogenic sulfate, black carbon (BC), and organic carbon (OC) aerosols over China are very different between the two models. The emission and deposition results, which are closely related to the model-assumed aerosol particle size distribution, indicate that the current aerosol size distribution used in the two models should be further improved. The differences in dust emission parameterizations also lead significant discrepancies in aerosol cycles and the dust emission scheme is an important factor determining the magnitudes of global and regional dust emission fluxes.

7.
Chin J Integr Med ; 26(5): 324-329, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32350801

RESUMO

Graft-versus-host disease (GVHD) is the most common complication after allogeneic hematopoietic stem cell transplantation, and also an important factor affecting the survival and quality of life in patients after transplantation. Currently, immunosuppressive therapy is commonly used for GVHD, but the curative effect is not ideal. How to effectively prevent and treat GVHD is one of the difficulties to be solved urgently in the field of transplantation. In this paper, we summarize the latest progress in pathogenesis, prevention and treatment of GVHD with Chinese medicine (CM). We hope it will provide ideas and methods for exploring the mechanism and establishing a new comprehensive therapy for GVHD with CM.


Assuntos
Doença Enxerto-Hospedeiro/tratamento farmacológico , Doença Enxerto-Hospedeiro/etiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Medicina Tradicional Chinesa , Aloenxertos , Humanos , Qualidade de Vida
8.
Chin J Integr Med ; 25(12): 902-910, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31802424

RESUMO

OBJECTIVE: To investigate the potential efficacy of panaxadiol saponins component (PDS-C) in the treatment of aplastic anemia (AA) model mice. METHODS: Totally 70 mice were divided into 7 groups as follows: normal, model, low-, medium-, high-dose PDS-C (20, 40, 80 mg/kg, namely L-, M-, H-PDS-C), cyclosporine (40 mg/kg), and andriol (25 mg/kg) groups, respectively. An immune-mediated AA mouse model was established in BALB/c mice by exposing to 5.0 Gy total body irradiation at 1.0 Gy/min, and injecting with lymphocytes from DBA mice. On day 4 after establishment of AA model, all drugs were intragastrically administered daily for 15 days, respectively, while the mice in the normal and model groups were administered with saline solution. After treatment, the peripheral blood counts, bone marrow pathological examination, colony forming assay of bone marrow culture, T lymphocyte subpopulation analysis, as well as T-bet, GATA-3 and FoxP3 proteins were detected by flow cytometry and Western blot. RESULTS: The peripheral blood of white blood cell (WBC), platelet, neutrophil counts and hemoglobin (Hb) concentration were significantly decreased in the model group compared with the normal group (all P<0.01). In response to 3 dose PDS-C treatment, the WBC, platelet, neutrophil counts were significantly increased at a dose-dependent manner compared with the model group (all P<0.01). The myelosuppression status of AA was significantly reduced in M-, H-PDS-C groups, and hematopoietic cell quantity of bone marrow was more abundant than the model group. The colony numbers of myeloid, erythroid and megakaryocytic progenitor cells in the model group were less than those of the normal mice in bone marrow culture, while, PDS-C therapy enhanced proliferation of hematopoietic progenitor cells by significantly increasing colony numbers (all P<0.01). Furthermore, PDS-C therapy increased peripheral blood CD3+ and CD3+CD4+ cells and reduced CD3+CD8+ cells (P<0.05 or P<0.01). Meanwhile, PDS-C treatment at medium- and high doses groups also increased CD4+CD25+FoxP3+ cells, downregulated T-bet protein expression, and upregulated GATA-3 and FoxP3 protein expressions in spleen cells (P<0.05). CONCLUSION: PDS-C possesses dual activities, promoting proliferation hematopoietic progenitor cells and modulating T lymphocyte immune functions in the treatment of AA model mice.


Assuntos
Anemia Aplástica/tratamento farmacológico , Ginsenosídeos/farmacologia , Hematopoese/efeitos dos fármacos , Panax , Saponinas/farmacologia , Linfócitos T/efeitos dos fármacos , Anemia Aplástica/sangue , Animais , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos BALB C
9.
Chin J Integr Med ; 25(7): 483-489, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31278626

RESUMO

Chronic primary immune thrombocytopenia (CITP) is the most common acquired autoimmune disease that seriously threaten the physical and mental health of patients. Compared with Western medicine treatment, the intervention and treatment of Chinese medicine (CM) has shown certain therapeutic advantages. This paper reviewed the new pathogenesis progress on T cell immune abnormality in CITP, and CM studies on interferes effects of cellular immune regulation of CITP in recent years. Qi deficiency failing to control blood and internal obstruction of blood stasis are the two common types of CM syndromes in CITP patients, the corresponding treatments include invigorating Pi (Spleen), supplementing qi, activating blood, as well as tonifying qi and activating yang, regulating Gan (Liver) to invigorate Pi. The authors also mentioned the problems in the research field of CM for CTIP treatment, and put forward new ideas for the research in the future.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Imunidade Celular , Púrpura Trombocitopênica Idiopática/tratamento farmacológico , Púrpura Trombocitopênica Idiopática/imunologia , Pesquisa , Hemorragia/tratamento farmacológico , Humanos
10.
Chin J Integr Med ; 23(5): 370-375, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27465424

RESUMO

OBJECTIVE: To investigate the potential effect of pure total flavonoids from Citrus paradisi Macfad peel (PTFC) on the proliferation of human myeloid leukemia cells Kasumi-1, HL-60 and K562, and the underlying mechanisms. METHODS: PTFC was extracted from Citrus paradisi Macfad peel and was identified by high performance liquid chromatography. The effect of PTFC on the proliferation and apoptosis of leukemia cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, fluorescent microscopy and flow cytometry, respectively. The effect of PTFC on the expression levels of apoptosis-related regulators was determined by Western blot assay. RESULTS: Treatment with PTFC inhibited leukemia cell proliferation in a dose- and time-dependent manner and triggered Kasumi-1 cell apoptosis. Treatment with PTFC significantly increased the levels of activated poly adenosine diphosphate-ribosepolymerase and caspase-3/-9, but reduced the levels of Mcl-1 expression in Kasumi-1 cells. However, PTFC did not obviously induce HL-60 cell apoptosis. CONCLUSION: PTFC inhibited leukemia cell proliferation and induced their apoptosis by modulating apoptosisrelated regulator expression in leukemia cells in vitro.


Assuntos
Apoptose/efeitos dos fármacos , Citrus paradisi/química , Flavonoides/uso terapêutico , Leucemia/tratamento farmacológico , Leucemia/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Flavanonas/farmacologia , Flavanonas/uso terapêutico , Flavonoides/farmacologia , Hesperidina/análogos & derivados , Hesperidina/farmacologia , Hesperidina/uso terapêutico , Humanos , Linfócitos/efeitos dos fármacos
11.
Environ Pollut ; 195: 319-29, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25017412

RESUMO

A new global aerosol assimilation system adopting a more complex icosahedral grid configuration is developed. Sensitivity tests for the assimilation system are performed utilizing satellite retrieved aerosol optical depth (AOD) from the Moderate Resolution Imaging Spectroradiometer (MODIS), and the results over Eastern Asia are analyzed. The assimilated results are validated through independent Aerosol Robotic Network (AERONET) observations. Our results reveal that the ensemble and local patch sizes have little effect on the assimilation performance, whereas the ensemble perturbation method has the largest effect. Assimilation leads to significantly positive effect on the simulated AOD field, improving agreement with all of the 12 AERONET sites over the Eastern Asia based on both the correlation coefficient and the root mean square difference (assimilation efficiency). Meanwhile, better agreement of the Ångström Exponent (AE) field is achieved for 8 of the 12 sites due to the assimilation of AOD only.


Assuntos
Aerossóis/análise , Poluentes Atmosféricos/análise , Monitoramento Ambiental/métodos , Modelos Químicos , Imagens de Satélites , Poluição do Ar/estatística & dados numéricos , Ásia
12.
Zhonghua Xue Ye Xue Za Zhi ; 34(4): 337-40, 2013 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-23668208

RESUMO

OBJECTIVE: To explore the proliferation inhibition and apoptosis effects of polysaccharides extracts from Hedyotis diffusa (PEHD) on multiple myeloma (MM) cell line RPMI 8226 cells in vitro, so as to provide experimental theory for the clinical application in the treatment of MM. METHODS: MTT assay was used to examine the effects of PEHD on cell growth. The apoptotic cells were analyzed by flow cytometry with AnnexinⅤ/PI staining. Hoechst staining was used to observe the morphological changes of RPMI 8226 cell apoptosis. The expression levels of caspase-3,-8,-9, PARP, nucleoprotein NF-κB protein and other channel protein were assayed by Western blotting method. RESULTS: The growth of RPMI 8226 cells were suppressed after treatment with PEHD, the highest inhibition rate reached to 92.3%, the results in the doses from 1 to 4 mg/ml showed a dose-and-time-dependent manner. The proportion of apoptotic cells in 1, 2 and 3 mg/ml PEHD treatment groups for 24 h were 22.52%, 62.31% and 69.94%, respectively, and significantly higher than that of control 8.93%. After treated with PEHD, apoptotic body appeared in RPMI 8226 cells nucleus and the number of apoptotic body increased in a dose-dependent manner. With the increasing of PEHD concentration, the expression of caspase-8,-9,-3 and PARP protein increased. The expression of Mcl-1, Bcl-xl, Bid and Bim protein decreased gradually, but the expression of Bax, Bak and Bad protein increased, and the expression of p-AKT protein (60 kDa) and NF-κB obviously decreased. CONCLUSION: PEHD could inhibited the growth of RPMI 8226 cells and displayed a dose-and-time-dependent manner, its mechanism may involve cell apoptosis induction, which was associated with the activation of caspase-8, caspase-9, and caspase-3 protein and the down-regulation of p-AKT and NF-κB protein expression.


Assuntos
Apoptose/efeitos dos fármacos , Hedyotis/química , Mieloma Múltiplo/patologia , Polissacarídeos/farmacologia , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Mieloma Múltiplo/metabolismo , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo
13.
Zhongguo Zhen Jiu ; 28(9): 673-4, 2008 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-18822985

RESUMO

OBJECTIVE: To compare therapeutic effects of acupuncture on complicated facial paralysis in the patient with HIV/AIDS and in the patient with no HIV/AIDS. METHODS: The observation group of 31 cases of facial paralysis with positive HIV/AIDS and the control group of 30 cases of facial paralysis with negative HIV/AIDS were treated with acupuncture at Yifeng (TE 17), Xiaguan (ST 7), Jiache (ST 6), Taiyang (EX-HN 5), Dicang (ST 4), Yuyao (EX-HN 4), Cuanzhu (BL 2), Yingxiang (LI 20), etc. and with individual treatment according to different periods of disease. The difference of therapeutic effects between the two groups were observed. RESULTS: The effective rate was 83.9% in the observation group and 96.7% in the control group, with no significant difference between the two groups; while the cured rate was 9.7% in the observation group and 73.3% in the control group with a very significant difference between the two groups (P < 0.01). CONCLUSION: Acupuncture with individual treatment has a satisfactory therapeutic effect on complicated facial paralysis in the patient of HIV/AIDS.


Assuntos
Síndrome da Imunodeficiência Adquirida/complicações , Terapia por Acupuntura , Paralisia Facial/terapia , Adolescente , Adulto , Feminino , Humanos , Masculino , Medicina Tradicional Chinesa
14.
Di Yi Jun Yi Da Xue Xue Bao ; 25(11): 1348-51, 2005 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-16305953

RESUMO

OBJECTIVE: To construct eukaryotic expression vectors of two mutants of hypoxia-inducible factor-1 (HIF-1alpha) and study their expressions in human microvascular endothelial cells (HMVECs). METHODS: Site-directed mutagenesis was performed to induce the mutation of the codons for the residue Pro564 (ccc) in HIF-1alpha into gcc (Ala) in pcDNA3.1(+)-HIF-1alphato obtain single-site-mutated vector pcDNA3.1(+)-HIF-1alpha-564Ala, which was then subjected to a second site-directed mutagenesis to convert the codons for Asn803 into that of Ala (gct) to acquire double-site-mutated pcDNA3.1(+)-HIF-1alpha-564Ala-803Ala. After lipofectin-mediated transient transformation of HMVECs with the 3 recombinant plasmids including the two plasmids containing the mutations and the one without mutation, respectively, the expression levels of HIF-1alpha mRNA and protein were determined using RT-PCR, immunofluorescent staining and Western blotting. RESULTS: DNA sequence analysis demonstrated success of the two-step mutagenesis and the two plasmids of pcDNA3.1+-HIF-1alpha-564Ala and pcDNA3.1(+)-HIF-1alpha-564Ala- 803Ala were obtained, both of which could produce HIF-1alpha protein resistant to oxidation degradation in HMVECs as compared with the non-mutated one. CONCLUSION: The recombinant plasmids pcDNA3.1(+)-HIF-1alpha-564Ala and pcDNA3.1(+)-HIF-1alpha- 564Ala-803Ala have been successfully constructed with efficient expressions in HMVECs.


Assuntos
Endotélio Vascular/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/biossíntese , Mutação Puntual , Clonagem Molecular , Endotélio Vascular/citologia , Células Eucarióticas/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Mutagênese Sítio-Dirigida , Neovascularização Fisiológica , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA
15.
Di Yi Jun Yi Da Xue Xue Bao ; 23(11): 1134-6, 2003 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-14625170

RESUMO

OBJECTIVE: To construct the eukaryotic expression vector for human hypoxia-inducible factor-1alpha (HIF-1alpha) gene and examine its expression. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on the total RNA extracted from HT29 cells to obtain the cDNA of HIF-1alpha, which was inserted into T vector pUC18. DNA sequencing was performed before the amplified products were cloned into the eukaryotic expression vector pcDNA3.1+ identified by endonuclease digestion. This recombinant vector was transfected into HEK293 cells by means of liposome and its expression examined. RESULTS: The amplified products were confirmed as the cDNA of HIF-1alpha by DNA sequencing, and pcDNA3.1+ -HIF-1alpha obtained was verified by endonuclease digestion, being capable of expression in HEK293 cells. CONCLUSION: We have successfully constructed the eukaryotic expression vector for HIF-1alpha, which can be expressed in HEK293 cells.


Assuntos
DNA Complementar/genética , Vetores Genéticos , Fatores de Transcrição/genética , Clonagem Molecular , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA
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