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1.
Int J Mol Sci ; 23(19)2022 Sep 22.
Article in English | MEDLINE | ID: mdl-36232459

ABSTRACT

A great paradigm for foremost food packaging is to use renewable and biodegradable lignocellulose-based materials instead of plastic. Novel packages were successfully prepared from the cellulose paper by coating a mixture of polylactic acid (PLA) with cinnamaldehyde (CIN) as a barrier screen and nano silica-modified stearic acid (SA/SiO2) as a superhydrophobic layer. As comprehensively investigated by various tests, results showed that the as-prepared packages possessed excellent thermal stability attributed to inorganic SiO2 incorporation. The excellent film-forming characteristics of PLA improved the tensile strength of the manufactured papers (104.3 MPa) as compared to the original cellulose papers (70.50 MPa), enhanced by 47.94%. Benefiting from the rough nanostructure which was surface-modified by low surface energy SA, the contact angle of the composite papers attained 156.3°, owning superhydrophobic performance for various liquids. Moreover, the composite papers showed excellent gas, moisture, and oil bacteria barrier property as a result of the reinforcement by the functional coatings. The Cobb300s and WVP of the composite papers were reduced by 100% and 88.56%, respectively, and their antibacterial efficiency was about 100%. As the novel composite papers have remarkable thermal stability, tensile strength, and barrier property, they can be exploited as a potential candidate for eco-friendly, renewable, and biodegradable cellulose paper-based composites for the substitute of petroleum-derived packages.


Subject(s)
Food Packaging , Petroleum , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Cellulose/chemistry , Hydrophobic and Hydrophilic Interactions , Plastics , Polyesters , Silicon Dioxide
2.
J Clin Lab Anal ; 36(7): e24508, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35689566

ABSTRACT

BACKGROUND: Arctigenin (ATG) is the active ingredient of the Chinese herbal medicine Arctium lappa, with anti-inflammatory and antioxidant effects. Excessive inflammation and cell apoptosis are important causes of intervertebral disc degeneration (IDD). Hence, this study probed into the possible role of ATG in IDD. METHODS: Interleukin (IL)-1ß (10 ng/ml) was adopted to induce human nucleus pulposus cells (HNPCs) as a cell model for IDD. The effects of different concentrations of ATG (0, 2, 5, 10, 20, 50 µmol/L) on the viability of HNPCs and effects of ATG (10, 50 µmol/L) on the viability of IL-1ß-induced HNPCs were detected by cell counting kit-8 (CCK-8). After IL-1ß-induced HNPCs were transfected with miR-483-3p inhibitor and/or treated with ATG, cell viability and apoptosis were determined by CCK-8 and flow cytometry; the expressions of miR-483-3p, extracellular matrix (ECM)-related genes, and inflammation-related genes were measured by quantitative real time polymerase chain reaction (qRT-PCR), and expressions of ECM/apoptosis/NF-κB pathway-related proteins were quantified by Western blot. RESULTS: ATG had no significant effect on the viability of HNPCs but could promote the viability of IL-1ß-induced HNPCs. ATG inhibited apoptosis, ECM degradation, inflammation, and activation of NF-κB pathway in HNPCs induced by IL-1ß, but promoted the expression of miR-483-3p. MiR-483-3p inhibitor reversed the above-mentioned regulatory effects of ATG. CONCLUSION: Arctigenin suppresses apoptosis, ECM degradation, inflammation, and NF-κB pathway activation in HNPCs by up-regulating miR-483-3p.


Subject(s)
Furans , Intervertebral Disc Degeneration , Lignans , MicroRNAs , Nucleus Pulposus , Apoptosis/genetics , Cells, Cultured , Extracellular Matrix/metabolism , Extracellular Matrix Proteins/genetics , Furans/pharmacology , Humans , Inflammation/genetics , Inflammation/metabolism , Intervertebral Disc Degeneration/drug therapy , Intervertebral Disc Degeneration/genetics , Lignans/pharmacology , MicroRNAs/genetics , MicroRNAs/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Nucleus Pulposus/metabolism
3.
Brachytherapy ; 19(1): 81-89, 2020.
Article in English | MEDLINE | ID: mdl-31653566

ABSTRACT

OBJECTIVES: The objective of this study was to evaluate the efficacy and safety of CT-guided radioactive 125I seed implantation as a salvage treatment for locally recurrent head and neck soft tissue sarcoma (HNSTS) after surgery and external beam radiotherapy. METHODS AND MATERIALS: From December 2006 to February 2018, 25 patients with locally recurrent HNSTS after surgery and external beam radiotherapy were enrolled. All the patients successfully underwent CT-guided 125I seed implantation. The primary end points included the objective response rate (ORR) and local progression-free survival (LPFS). The secondary end points were survival (OS) and safety profiles. RESULTS: After 125I seed implantation, the ORR was 76.0%. The 1-, 3-, and 5-year LPFS rates were 65.6%, 34.4%, and 22.9%, respectively, with the median LPFS of 16.0 months. The 1-, 3-, and 5-year OS rates were 70.8%, 46.6%, and 34.0%, respectively, with the median OS of 28.0 months. Furthermore, univariate analyses showed that the recurrent T stage and histological grade were prognostic factors of LPFS, whereas only the histological grade was a predictor of OS. The major adverse events were skin/mucosal toxicities, which were generally of lower grade (≤Grade 2) and were well tolerated. CONCLUSIONS: Radioactive 125I seed implantation could be an effective and safe alternative treatment for locally recurrent HNSTS after failure of surgery and radiotherapy. Recurrent T stage and histological grade were the main factors influencing the efficacy.


Subject(s)
Brachytherapy/methods , Head and Neck Neoplasms/radiotherapy , Neoplasm Recurrence, Local/radiotherapy , Salvage Therapy/methods , Sarcoma/radiotherapy , Soft Tissue Neoplasms/radiotherapy , Adult , Brachytherapy/adverse effects , Female , Head and Neck Neoplasms/pathology , Head and Neck Neoplasms/surgery , Humans , Iodine Radioisotopes , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Progression-Free Survival , Radiotherapy, Adjuvant , Radiotherapy, Image-Guided/adverse effects , Radiotherapy, Image-Guided/methods , Sarcoma/pathology , Sarcoma/surgery , Soft Tissue Neoplasms/pathology , Soft Tissue Neoplasms/surgery , Survival Rate , Tomography, X-Ray Computed , Young Adult
4.
Int J Radiat Oncol Biol Phys ; 103(3): 638-645, 2019 03 01.
Article in English | MEDLINE | ID: mdl-30391521

ABSTRACT

PURPOSE: To analyze the efficacy and safety of radioactive I-125 seed implantation in the treatment of recurrent head and neck tumors after radiation therapy. METHODS AND MATERIALS: The data of 101 patients with recurrent head and neck cancer after radiation therapy who received computed tomography guided radioactive I-125 seed implantation were analyzed. The median previous cumulative external irradiation dose was 66 Gy, and the median dose to 90% of the target volume (D90) after operation was 117 Gy. The short-term efficacy was evaluated by Response Evaluation Criteria in Solid Tumors version 1.1, and the adverse event was evaluated by Common Terminology Criteria for Adverse Events version 4.0. RESULTS: The 5-year local control rate was 26.6%, and the 5-year overall survival rate was 15.5%. Univariate analysis showed that factors related to local control rate included age, pathologic type, implantation site, lesion volume, and D90. The 5-year local control rate was 11.5% (2-year) if D90 was <120 Gy and 44.2% if D90 was ≥120 Gy (P = .001). Multivariate analysis showed that pathologic type, lesion volume, and D90 were independent factors related to local control (P = .002, 0, .014, respectively); Karnofsky performance status and lesion volume were independent factors associated with survival (P = .021 and 0, respectively). For the side effects, there were 26 cases of skin or mucosa ulceration (25.7%), 14 cases of pain (13.9%), and 2 cases of dry mouth (2%). The correlation between toxicity and dose had not been found. CONCLUSIONS: Radioactive I-125 seed implantation in the treatment of recurrent head and neck cancer after radiation therapy showed acceptable efficacy and safety. Nonsquamous carcinoma, small lesion volume, and high dose (D90) were correlated with better local control.


Subject(s)
Head and Neck Neoplasms/diagnostic imaging , Head and Neck Neoplasms/radiotherapy , Iodine Radioisotopes/chemistry , Neoplasm Recurrence, Local , Radiotherapy/methods , Adenocarcinoma/diagnostic imaging , Adenocarcinoma/radiotherapy , Adult , Aged , Aged, 80 and over , Brachytherapy , Carcinoma, Squamous Cell/diagnostic imaging , Carcinoma, Squamous Cell/radiotherapy , Female , Humans , Male , Middle Aged , Multivariate Analysis , Prognosis , Radiotherapy Planning, Computer-Assisted/methods , Retrospective Studies , Sarcoma/diagnostic imaging , Sarcoma/radiotherapy , Tomography, X-Ray Computed , Treatment Outcome , Young Adult
5.
Korean J Intern Med ; 34(5): 1078-1090, 2019 Sep.
Article in English | MEDLINE | ID: mdl-29432674

ABSTRACT

BACKGROUND/AIMS: Evidence suggests that Shen-Kang (SK), a traditional Chinese herbal medicine, protects against various types of renal injury. In this study, we evaluated whether SK treatment confers renoprotection in a rat model of chronic tacrolimus (TAC) nephropathy. METHODS: Rats were treated daily with TAC (1.5mg/kg, subcutaneously) and SK (450 mg/kg, intravenously) for 4 weeks. The effects of SK on TAC-induced renal injury were assessed by measuring renal function, urine albumin excretion, histopathology, inflammatory cell infiltration, expression of profibrotic (transforming growth factor ß1 [TGF-ß1] and TGF-ß inducible gene-h3 [ßig-h3]) and proinflammatory cytokines, oxidative stress, and apoptotic cell death. RESULTS: Administration of SK preserved glomerular integrity (fractional mesangial area and Wilms tumor 1-positive glomeruli), attenuated tubulointerstitial fibrosis, and reduced the number of ectodermal dysplasia 1-positive cells, and this was paralleled by improved urine albumin excretion and renal dysfunction. At the molecular level, SK treatment suppressed expression of TGF-ß1/Smad2/3, ßig-h3, and proinflammatory cytokines. Oxidative stress and apoptotic cell death were significantly decreased with SK treatment, and apoptosis-related genes were regulated toward cell survival (active caspase-3 and the B-cell lymphoma-2/Bcl2-associated X [Bcl-2/Bax] ratio). CONCLUSION: SK protects against TAC-induced renal injury.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Kidney Diseases/prevention & control , Kidney/drug effects , Protective Agents/pharmacology , Tacrolimus , Animals , Apoptosis/drug effects , Apoptosis Regulatory Proteins/metabolism , Cytokines/metabolism , Cytoprotection , Disease Models, Animal , Extracellular Matrix Proteins/metabolism , Kidney/metabolism , Kidney/pathology , Kidney Diseases/chemically induced , Kidney Diseases/metabolism , Kidney Diseases/pathology , Male , Oxidative Stress/drug effects , Rats, Sprague-Dawley , Signal Transduction , Transforming Growth Factor beta/metabolism , Transforming Growth Factor beta1/metabolism
6.
Clin Nutr ; 38(1): 159-164, 2019 02.
Article in English | MEDLINE | ID: mdl-29395372

ABSTRACT

OBJECTIVE: Sarcopenia, an age-related decline of muscle mass, strength, and physical function, was associated with falls, frailty, and poor quality of life. The aim of the current study is to examine the effect of nutritional supplement containing whey protein, vitamin D and E on measures of sarcopenia. METHODS: A total of 60 sarcopenic older adult subjects participated in the current randomized, double-blind, placebo-controlled (iso-caloric control product) trial for 6 months. Muscle mass [Relative skeletal mass index (RSMI) measured by bioimpedance analysis (BIA)], muscle strength (handgrip strength), physical function (6-m gait speed, chair stand test, and timed-up-and-go test, TUG), quality of life (measured by Short-Form 36-Item Health Survey, SF-36), and blood biochemical indexes were measured before and after the 6-month intervention. RESULTS: Compared to placebo group, nutritional supplementation improves RSMI (mean difference: 0.18 kg/m2, 95%CI: 0.01-0.35, P = 0.040), handgrip strength (mean difference: 2.68 kg, 95%CI: 0.71-4.65, P = 0.009), SF-36 mental component summary (SF-36 MCS) (mean difference: 11.26, 95%CI: 3.86-18.65, P = 0.004), SF-36 physical component summary (SF-36 PCS) (mean difference: 20.21, 95%CI: 11.30-29.12, P < 0.001), serum IGF-1 (mean difference: 14.34 ng/mL, 95%CI: 2.06-26.73), IL-2 (mean difference: -575.32 pg/mL, 95%CI: -1116.94 âˆ¼ -33.70, P = 0.038), serum vitamin D3 (mean difference: 11.01 ng/mL, 95%CI: 6.44-15,58, P < 0.001), and serum vitamin E (mean difference: 4.17 ng/L, 95%CI: 1.89-6.45, P = 0.001). CONCLUSION: The current study demonstrated that the combined supplementation of whey protein, vitamin D and E can significantly improve RSMI, muscle strength, and anabolic markers such as IGF-I and IL-2 in older adults with sarcopenia. Further larger well-designed studies are warranted to evaluate whether long-term whey protein supplementation can blunt the declines of muscle function and mass in older adults with sarcopenia.


Subject(s)
Dietary Supplements , Muscle Strength/drug effects , Quality of Life , Sarcopenia/diet therapy , Vitamin D/therapeutic use , Vitamin E/therapeutic use , Whey Proteins/therapeutic use , Aged , Aged, 80 and over , Double-Blind Method , Female , Humans , Male , Middle Aged , Muscle, Skeletal/drug effects , Vitamins/therapeutic use
7.
Article in English | WPRIM | ID: wpr-919143

ABSTRACT

BACKGROUND/AIMS@#Evidence suggests that Shen-Kang (SK), a traditional Chinese herbal medicine, protects against various types of renal injury. In this study, we evaluated whether SK treatment confers renoprotection in a rat model of chronic tacrolimus (TAC) nephropathy.@*METHODS@#Rats were treated daily with TAC (1.5mg/kg, subcutaneously) and SK (450 mg/kg, intravenously) for 4 weeks. The effects of SK on TAC-induced renal injury were assessed by measuring renal function, urine albumin excretion, histopathology, inflammatory cell infiltration, expression of profibrotic (transforming growth factor β1 [TGF-β1] and TGF-β inducible gene-h3 [βig-h3]) and proinflammatory cytokines, oxidative stress, and apoptotic cell death.@*RESULTS@#Administration of SK preserved glomerular integrity (fractional mesangial area and Wilms tumor 1-positive glomeruli), attenuated tubulointerstitial fibrosis, and reduced the number of ectodermal dysplasia 1-positive cells, and this was paralleled by improved urine albumin excretion and renal dysfunction. At the molecular level, SK treatment suppressed expression of TGF-β1/Smad2/3, βig-h3, and proinflammatory cytokines. Oxidative stress and apoptotic cell death were significantly decreased with SK treatment, and apoptosis-related genes were regulated toward cell survival (active caspase-3 and the B-cell lymphoma-2/Bcl2-associated X [Bcl-2/Bax] ratio).@*CONCLUSIONS@#SK protects against TAC-induced renal injury.

8.
J Cancer Res Ther ; 14(7): 1492-1496, 2018.
Article in English | MEDLINE | ID: mdl-30589028

ABSTRACT

BACKGROUND: I-125 seed implantation has been widely applied in the local treatment of advanced malignant tumor. It has the advantages of providing a high dose of treatment to the target sites and low dose to normal tissues. It has been mostly applied as palliative treatment for recurrences in advanced malignant tumor (except for prostate cancer), suppressing tumor development and improving the quality of life of patients. OBJECTIVE: The objective of this study was to investigate changes in quality of life for patients with advanced malignant tumor after receiving I-125 seed implantation using a three-dimensional (3D)-printed individualized template and computed tomography (CT) guidance. MATERIALS AND METHODS: In this prospective study, convenience sampling was applied for patients with advanced tumors attending a tertiary hospital. The European Organization for Research on Treatment of Cancer Quality of Life Questionnaire-C30 was involved to assess quality of life. Patients completed the questionnaire before and 24 h after seed implantation. The questionnaire of 1 and 3 months after seed implantation was completed by telephonic follow-up. RESULTS: A total of 42 patients were included (24 males and 18 females), with an average age of 58.86 ± 14.13 years (ranged 25-91 years). The average scale score after seed implantation was higher than that of before implantation. The order was the average scale score 1 month after seed implantation >3 months after seed implantation >24 h after seed implantation. CONCLUSION: The results suggested that the quality of life could be improved with I-125 seed implantation using a 3D-printed individualized template under CT guidance in patients with the advanced malignant tumor.


Subject(s)
Brachytherapy , Iodine Radioisotopes , Neoplasms/diagnosis , Neoplasms/radiotherapy , Quality of Life , Radiotherapy Planning, Computer-Assisted , Radiotherapy, Image-Guided , Tomography, X-Ray Computed , Adult , Aged , Aged, 80 and over , Brachytherapy/methods , Female , Humans , Male , Middle Aged , Neoplasm Staging , Neoplasms/mortality , Radiotherapy, Image-Guided/methods , Treatment Outcome
9.
J Orthop Surg Res ; 10: 103, 2015 Jul 03.
Article in English | MEDLINE | ID: mdl-26138716

ABSTRACT

BACKGROUND: Elcatonin (ECT) is used to prevent and treat osteoporosis. However, little is known about its effect on the disuse osteoporosis (DOP). The aim of this study is to evaluate the effect of ECT on DOP caused by fracture fixation. METHODS: Forty-five male Sprague-Dawley (SD) rats, aged 6 weeks, were randomly allocated into three groups: the control group without surgery and elcatonin treatment (CTR, n = 15), the surgery group without elcatonin treatment (SUR, n = 15), and the surgery group which received elcatonin subcutaneously (SUR + ECT, n = 15). Surgery was produced by cutting the midshaft of the right femur transversely, fixing with stainless intramedullary needle, and immobilizing the right leg. All the proximal tibias from the random five rats in each group were harvested and investigated by evaluating bone mineral density (BMD), X-ray images, and histological staining respectively at the 4th, 8th, and 12th weeks after surgery. RESULTS: Both of the SUR and SUR + ECT groups obviously exhibited lower BMD values compared to the CTR group; however, the SUR + ECT group showed significantly higher BMD values (p < 0.001, p < 0.05, and p < 0.05) than the SUR group at each time point after surgery. Moreover, similar changes were observed between these groups when examining the radiographs and hematoxylin and eosin (HE) staining. CONCLUSIONS: Elcatonin attenuates disuse osteoporosis after fractures in rats, which may provide a new avenue to prevent and treat disuse osteoporosis after surgery in clinic.


Subject(s)
Calcitonin/analogs & derivatives , Fracture Fixation/adverse effects , Osteoporosis/prevention & control , Animals , Bone Density , Bone and Bones/diagnostic imaging , Bone and Bones/pathology , Calcitonin/therapeutic use , Drug Evaluation, Preclinical , Male , Osteoporosis/etiology , Osteoporosis/pathology , Radiography , Random Allocation , Rats, Sprague-Dawley
10.
Int J Med Mushrooms ; 13(5): 441-8, 2011.
Article in English | MEDLINE | ID: mdl-22324409

ABSTRACT

The immunomodulatory effect of GLIS (Lingzhi or Reishi medicinal mushroom Ganoderma lucidum immunomodulating substance) on macrophages has been investigated as part of ongoing research into the anticancer properties of this mushroom. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated BMMs were enlarged and formed pseudopodia. Exposure of BMMs to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1ß, IL-6, IL-12p35, IL-12p40, IL-18, and TNF-α gene expression and levels of TNF-α, IL-1ß, and IL-12 secretion. Our data indicate that GLIS activates the immune system by modulating cytokine production.


Subject(s)
Bone Marrow Cells/drug effects , Proteoglycans/chemistry , Proteoglycans/pharmacology , Reishi/chemistry , Animals , Cytokines/genetics , Cytokines/metabolism , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Gene Expression Regulation/immunology , Immunomodulation , Mice , Mice, Inbred BALB C , Respiratory Burst/drug effects , Reverse Transcriptase Polymerase Chain Reaction , Specific Pathogen-Free Organisms
11.
Zhonghua Yi Xue Za Zhi ; 88(24): 1711-4, 2008 Jun 24.
Article in Chinese | MEDLINE | ID: mdl-19024545

ABSTRACT

OBJECTIVE: To investigate the neuroprotective effect of epigallocatechin gallate(EGCG), the main extract from green tea, on the oxidative-stress-injured retinal ganglion cells. METHODS: Rat retinal ganglion cells (RGC-5) were cultured into 3 groups (normal control; H2O2; H2O2 + EGCG or Trolox or NU1025). In-situ TUNEL was used to detect the apoptosis of the RGC-5 cells. Dihydroethidium (DHE) assay was used to observe the intracellular ROS generation. The activation of nuclear enzyme, PARP-1 was quantitatively detected by Western blot and the cell viability was measured by MT method. RESULTS: Hydrogen peroxide reduced RGC-5 cell viability in a time-concentration-dependent manner. The treatment of 500 micromol/L H2O2 for 24 hours reduced RGC-5 cell viability by about 50% of control. Hydrogen peraoxide caused apoptosis of the RGC-5 cell, obviously increased intracellular ROS generation and up-regulated the PARP-1 expression. The pretreatment with EGCG was able to markedly reduce the number of apoptotic cells, attenuate intracellular ROS generation. Furthermore, MTT assay showed that the pretreatment with EGCG (50 micromol/L) increased the most cell viability to 87% of control, but pretreatment with Trolox (100 micromol/L) and NU1025 (100 micromol/L, a PARP-1 inhibitor) recovered the most cell viability to 62% and 71% of control respectively. CONCLUSION: EGCG is able to effectively protect retinal ganglion cell against oxidative-stressed injury and can be used as a very potential neuroprotective drug.


Subject(s)
Catechin/analogs & derivatives , Neuroprotective Agents/pharmacology , Oxidative Stress/drug effects , Retinal Ganglion Cells/drug effects , Animals , Apoptosis/drug effects , Catechin/pharmacology , Cell Survival/drug effects , Cells, Cultured , Hydrogen Peroxide/pharmacology , In Situ Nick-End Labeling , Rats , Reactive Oxygen Species/metabolism , Retinal Ganglion Cells/cytology , Retinal Ganglion Cells/metabolism , Tea/chemistry
12.
J Ethnopharmacol ; 112(3): 445-50, 2007 Jul 25.
Article in English | MEDLINE | ID: mdl-17524580

ABSTRACT

The immunomodulatory effect of Ganoderma lucidum immunomodulating substance (GLIS) on macrophages has been investigated as part of on-going research into the anti-cancer properties of Ganoderma lucidum. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated RAW264.7 macrophages were enlarged and formed pseudopodia. Exposure of RAW264.7 macrophages to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1beta, IL-12p35 and IL-12p40 gene expression. Our data indicate that GLIS activates the immune system by modulating cytokine production.


Subject(s)
Immunologic Factors/pharmacology , Macrophages/drug effects , Proteoglycans/pharmacology , Reishi/chemistry , Animals , Cell Line , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Gene Expression/drug effects , Immunologic Factors/immunology , Immunologic Factors/isolation & purification , Interleukin-12 Subunit p35/genetics , Interleukin-12 Subunit p40/genetics , Interleukin-1beta/genetics , L Cells , Luminescent Measurements/methods , Luminol/metabolism , Macrophages/cytology , Macrophages/metabolism , Mice , Mice, Inbred BALB C , Nitric Oxide/metabolism , Polysaccharides/pharmacology , Proteoglycans/immunology , Proteoglycans/isolation & purification , RNA, Messenger/genetics , RNA, Messenger/metabolism , Respiratory Burst/drug effects , Respiratory Burst/immunology , Reverse Transcriptase Polymerase Chain Reaction
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