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Improving cancer immunotherapy via co-delivering checkpoint blockade and thrombospondin-1 downregulator

Acta Pharmaceutica Sinica B 2023年第8期13卷 3503-3517页

作者： Qingqing Xiao Xiaotong Li Chang Liu Yuxin Jiang Yonglong He Wanting Zhang Helena SAzevedo Wei Wu Yuanzheng Xia Wei He School of Pharmacy China Pharmaceutical UniversityNanjing 211198China School of Engineering and Materials Science Institute of BioengineeringQueen Mary University of LondonLondon El 4NSUK School of Pharmacy Fudan UniversityShanghai 201203China Jiangsu Key Laboratory of Bioactive Natural Product Research and State Key Laboratory of Natural Medicines School of Traditional Chinese PharmacyChina Pharmaceutical UniversityNanjing 210009China

The use of checkpoint-blockade antibodies is still restricted in several malignancies due to the modest efficacy,despite considerable success in anti-tumor immunotherapy.The poor response of cancer cells to immune destruction is an essential contributor to the failure of checkpoint therapy.We hypothesized that combining checkpoint therapy with natural-product chemosensitizer could enhance immune response.Herein,a targeted diterpenoid derivative was integrated with the checkpoint blockade(anti-CTLA-4)to improve immunotherapy using thermo sensitive liposomes as carriers.In vivo,the liposomes enabled the co-delivery of the two drug payloads into the tumor.Consequently,the regulatory T cell proliferation was restrained,the cytotoxic T cell infiltration was enhanced,and the profound immunotherapeutic effect was achieved.In addition,the immunotherapeutic effect of another clinically used checkpoint antibody,anti-PD-1,also benefited from the diterpenoid derivative.Of note,our mechanism study revealed that the targeted diterpenoid derivative increased the sensitivity of cancer cells to immune attack via THBS1 downregulation and the resultant destruction of THBS1-CD47 interaction.Collectively,co-delivering THBS1 inhibitor and checkpoint blockade is promising to boost cancer immunotherapy.We first time discovered that THBS1 suppression could strengthen checkpoint therapy.

关键词： Immunotherapy Diterpenoid-based conjugate Checkpoint blockade thrombospondin-1 Co-delivery Liposomes

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Assessment for the minimal invasiveness of laparoscopic liver resection by interleukin-6 and thrombospondin-1

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World Journal of Hepatology 2022年第1期14卷 234-243页

作者： Takayoshi Kaida Hiromitsu Hayashi Hiroki Sato Shotaro Kinoshita Takashi Matsumoto Yuta Shiraishi Yuki Kitano Takaaki Higashi Katsunori Imai Yo-ichi Yamashita Hideo Baba Gastroenterological Surgery Graduate School of Life SciencesKumamoto UniversityKumamoto 860-8556Japan

BACKGROUND Laparoscopic surgery has been introduced as a minimally invasive technique for the treatment of various field.However,there are few reports that have scientifically investigated the minimally invasive nature of laparoscopic liver resection(LLR).AIM To investigate whether LLR is scientifically less invasive than open liver resection.METHODS During December 2011 to April 2015,blood samples were obtained from 30 patients who treated with laparoscopic(n=10,33%)or open(n=20,67%)partial liver resection for liver tumor.The levels of serum interleukin-6(IL-6)and plasma thrombospondin-1(TSP-1)were measured using ELISA kit at four time points including preoperative,immediate after operation,postoperative day 1(POD1)and POD3.Then,we investigated the impact of the operative approaches during partial hepatectomy on the clinical time course including IL-6 and TSP-1.RESULTS Serum level of IL-6 on POD1 in laparoscopic hepatectomy was significantly lower than those in open hepatectomy(8.7 vs 30.3 pg/mL,respectively)(P=0.003).Plasma level of TSP-1 on POD3 in laparoscopic hepatectomy was significantly higher than those in open hepatectomy(1704.0 vs 548.3 ng/mL,respectively)(P=0.009),and have already recovered to preoperative level in laparoscopic approach.In patients with higher IL-6 Levels on POD1,plasma level of TSP-1 on POD3 was significantly lower than those in patients with lower IL-6 Levels on POD1.Multivariate analysis showed that open approach was the only independent factor related to higher level of IL-6 on POD1[odds ratio(OR),7.48;95%confidence interval(CI):1.28-63.3;P=0.02].Furthermore,the higher level of serum IL-6 on POD1 was significantly associated with lower level of plasm TSP-1 on POD3(OR,5.32;95%CI:1.08-32.2;P=0.04)in multivariate analysis.CONCLUSION In partial hepatectomy,laparoscopic approach might be minimally invasive surgery with less IL-6 production compared to open approach.

关键词： Laparoscopic surgery Liver resection Hepatectomy Minimal invasiveness Interleukin-6 thrombospondin-1

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Pterostilbene supresses inflammation-induced melanoma metastasis by impeding neutrophil elastase-mediated thrombospondin-1 degradation

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Chinese Herbal Medicines 2023年第1期15卷 94-101页

作者： Dan Su Shan Xu Kailin Ji Hailing Xu Yan Li Zhisheng Zhang Yuqing Shen Gaoyang Chen Department of Pharmacy Changzhou No.2 People’s HospitalAffiliated Hospital of Nanjing Medical UniversityChangzhou 213004China School of Pharmacy Changzhou UniversityChangzhou 213164China Department of Oncology The Second People’s Hospital of TaizhouTaizhou 225300China

Objective: Chronic inflammation plays a fatal role in tumor metastasis. Pterostilbene(PTE) is a natural dimethylated analogue of resveratrol with anticancer and anti-inflammatory activities. This study aimed to investigate the inhibitory effect of PTE on inflammation-associated metastasis and explore the underlying mechanisms.Methods: Lipopolysaccharide(LPS)-induced lung inflammation and melanoma metastasis models were established in mice. After PTE treatment for four weeks, the organ index, histological changes, proinflammatory cytokines, and the expression and activity of neutrophil elastase(NE), a biomarker of neutrophil influx in the lungs, were analysed. Additionally, direct effects of PTE on NE-induced B16 cell migration were explored in wound healing and Transwell assays, and the expression of thrombospondin-1(TSP-1) and epithelial-mesenchymal transition(EMT) markers were also detected.Results: PTE obviously attenuated the LPS-induced metastasis of circulatory B16 cells to lungs by reducing the number of metastatic nodules on the lung surfaces and the lung weight/body weight ratio. PTE treatment also significantly reduced LPS-activated increase levels of tumor necrosis factor(TNF)-a and interleukin(IL)-6 in the lungs of tumor-bearing mice. In addition, increased expression and enzyme activity of NE and decreased expression of TSP-1 were observed, and these were blocked by PTE. In vitro, PTE at concentrations without cytotoxicity also markedly suppressed NE-triggered B16 cell migration, prevented NE-induced TSP-1 proteolysis and reversed the expression of vimentin, N-cadherin and Ecadherin.Conclusion: PTE could block inflammation-enhanced tumor metastasis, and the underlying mechanism might be associated with the inhibition of NE-mediated TSP-1 degradation.

关键词： inflammation melanoma metastasis neutrophil elastase pterostilbene thrombospondin-1

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Role of Elevated thrombospondin-1 in Kainic Acid-Induced Status Epilepticus

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Neuroscience Bulletin 2020年第3期36卷 263-276页

作者： Yurong Zhang Mengdi Zhang Wei Zhu Xiaohong Pan Qiaoyun Wang Xue Gao Chaoyun Wang Xiuli Zhang Yuxia Liu Shucui Li Hongliu Sun School of Pharmaceutical Sciences Binzhou Medical UniversityYantai 264003China Shandong Academy of Medical Sciences Jinan 250062China

Previous studies have suggested that thrombospondin-1(TSP-1) regulates the transforming growth factor beta 1(TGF-b1)/phosphorylated Smad2/3(p Smad2/3) pathway. Moreover, TSP-1 is closely associated with epilepsy. However, the role of the TSP-1-regulated TGFb1/p Smad2/3 pathway in seizures remains unclear. In this study, changes in this pathway were assessed following kainic acid(KA)-induced status epilepticus(SE) in rats.The results showed that increases in the TSP-1/TGF-b1/p Smad2/3 levels spatially and temporally matched the increases in glial fibrillary acidic protein(GFAP)/chondroitin sulfate(CS56) levels following KA administration.Inhibition of TSP-1 expression by small interfering RNA or inhibition of TGF-b1 activation with a Leu-Ser-Lys-Leu peptide significantly reduced the severity of KA-induced acute seizures. These anti-seizure effects were accompanied by decreased GFAP/CS56 expression and Smad2/3 phosphorylation. Moreover, inhibiting Smad2/3 phosphorylation with ponatinib or SIS3 also significantly reduced seizure severity, alongside reducing GFAP/CS56 immunoreactivity. These results suggest that the TSP-1-regulated TGF-b1/p Smad2/3 pathway plays a key role in KA-induced SE and astrogliosis, and that inhibiting this pathway may be a potential anti-seizure strategy.

关键词： Astrogliosis Status epilepticus Ponatinib thrombospondin-1

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Correlation between the expression of thrombospondin-1 and neovascularization in mucoepidermoid carcinoma

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Chinese Medical Journal 2008年第19期121卷 1875-1881页

作者： YANG Sen WANG Xiao-yi GUO Li-juan TANG Xiu-fa GAO Qing-hong XUAN Ming ZHANG Qiang TAN Ke WEN Yu-ming WANG Chang-mei XU Rui-an CAO Yu-tang State Key Laboratory of Oral Diseases Sichuan University Chengdu Sichuan 610041 China Department of Head and Neck Tumor Surgery West China College of Stomatology Sichuan University Chengdu Sichuan 610041 China Department of Health Statistics West China College of Public Health Sichuan University Chengdu Sichuan 610041 China

Background Researchers have recently demonstrated that thrombospondin-1 （TSP-1） has an important function in regulating neovascularization. Whether it inhibits or accelerates neovascularization, however, is still controversial. We found few reports about the correlation between TSP-1 and vascularization in mucoepidermoid carcinoma. In this research, the distribution and expression of TSP-1 in mucoepidermoid carcinoma were investigated. We also analyzed （1） the correlation between the expression of TSP-1 and microvessel density （MVD）, as an indicator of neovascularization activity, and （2） the effect of TSP-1 on neovascularization and tumor growth in the subcutaneous xenotransplanted model of mucoepidermoid carcinoma. Method （1） The sites and intensity of expression of TSP-1 and the MVD were analyzed in 45 cases of mucoepidermoid carcinoma after surgery by the method of streptavidin-peroxidase （SP） immunohistochemistry; and （2） recombinant human thrombospondin-1 （rhTSP-1） was injected twice a week for five consecutive weeks around the tumor in the subcutaneous xenotransplanted tumor model of mucoepidermoid carcinoma in nude mice. Each week, the tumor size was measured, in order to draw the growth curve of the xenotransplanted tumor model of mucoepidermoid carcinoma, and MVD was measured. Results （1） The positive expression of TSP-1 protein was 57.78% （26/45）. Most positive staining for TSP-1 was found in the cytoplasm of the cancer cells, while some staining occurred in the extracellular matrix. The mean MVD in 45 cases of m ucoepidermoid carcinoma was 58.17±19.77 per 100 visual fields. Tumors with a high expression of TSP-1 showed a low MVD value, and the TSP-1 immunocompetence and microvessel density showed a significant negative correlation （rs=-0.947, P 〈0.001）. （2） The xenotransplanted tumors with the injection doses of 1.25, 0.75 and 0.25 ug/ml respectively were 36.97%, 53.36% and 73.61% of the size of the control group （（451±92）, （651±113）, （898±86） and （1220±157） mm^3 respectively, F=-53.167, P 〈0.001）, an

关键词： mucoepidermoid carcinoma thrombospondin-1 recombinant human thrombospondin-1 microvessel density

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Protection of erythropoietin on experimental spinal cord injury by reducing the expression of thrombospondin-1 and transforming growth factor-β

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Chinese Medical Journal 2009年第14期122卷 1631-1635页

作者： FANG Xiang-qian FANG Mei FAN Shun-wu GU Chuan-long Department of Orthopaedics Sir Run Run Shaw Hospital School of Medicine Zhejiang University Hangzhou Zhejiang 310016 China Sir Run Run Shaw Hospital School of Medicine Zhejiang University Hangzhou Zhejiang 310016 China Department of Cell Biology and Anatomy School of Medicine Zhejiang University Hangzhou Zhejiang 310058 China

Background Erythropoietin （EPO） functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production and neuron protection. This study aimed to determine the neuron protective effect of erythropoietin on experimental rats enduring spinal cord injury （SCI） by assessing thrombospondin-1 （TSP-1） level and transforming growth factor-β （TGF-β） in the development of a rat model of SCI. Methods Sixty Sprague-Dawley rats were randomly assigned to three groups： sham operation control group, SCI group and EPO treatment group. By using a weight-drop contusion SCI model, the rats in the SCI group and EPO treatment group were sacrificed at 24 hours and 7 days subsequently. The Basso, Beattie, and Bresnahan （BBB） scores were examined for locomotor function. Pathological changes were observed after HE staining. The expressions of thrombospondin-2 （TSP-1） and TGF-β were determined by immunohistochemical staining and Western blotting. Results Slighter locomotor dysfunction was discovered and it was recovered abruptly as higher BBB scores were found in the EPO treatment group than in the SCI group （P 〈0.01）. Pathologically, progressive disruption of the dorsal white matter and regeneration of a few neurons were also observed in SCI rats. TSP-1 and TGF-β expression increased at 24 hours and 7 days after SCI in the injured segment, and it was higher in the SCI group than in the EPO treatment group. Spinal cord samples from the animals demonstrated a TSP-1 optical density of 112.2±6.8 and TSP-1 positive cells of 5.7±1.3 respectively. After injury, the TSP-1 optical density and cell number increased to 287.2±14.3/mm^2 and 23.2±2.6/mm^2 at 24 hours and to 232.1±13.2/mm^2 and 15.2±2.3/mm^2 at 7 days respectively. When EPO treated rats compared with the SCI rats, the TSP-1 optical density and cell number decreased to 213.1 ±11.6/mm^2 and 11.9±1.6/mm^2 at 24 hours and to 189.9±10.5/mm^2 and 9.3±1.5/mm^2 at 7 days, respectively （P 〈0.01）. In the SCI rats, the TGF-β optical density and positive neuron nu

关键词： thrombospondin-1 transforming growthfactor-β spinal cord injury erythropoietin

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thrombospondin-1 Levels in Patients with Coronary Heart Disease

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World Journal of Cardiovascular Diseases 2021年第6期11卷 277-291页

作者： Mohamed Elnoamany Ashraf Dawood Nahed Mohamed Momtaz Waleed Abdou Cardiology Department Faculty of Medicine Menoufia University Menoufia Egypt

Background: thrombospondin-1 (TSP1) is associated with atherosclerosis in animals with diabetes mellitus (DM), but the precise role of TSP-1 in human atherosclerosis remains unknown. Objectives: To investigate serum thrombospondin1 level in patients with coronary artery disease with and without type 2 DM and its relationship to coronary artery scoring systems. Methods: The study comprised 180 patients recruited from those underwent coronary angiography for suspected coronary artery disease (CAD) was approved by Institutional Review Board and Institutional Ethical Committee for Human Research of menoufia university hospital. They were divided according to presence of CAD and type 2 DM into 4 groups: Group I (n = 44 patients): Non diabetic subjects without CAD, Group II (n = 40 patients): Diabetic patients without CAD, Group III (n = 49 patients): Non diabetic patients with CAD and Group IV (n = 47 patients): Diabetic patients with CAD. Serum level of TSP-1 was measured in all groups and coronary artery scoring analysis was done. Results: Serum TSP-1 levels were higher in patients with CAD and DM than in other groups (P 0.01) while the levels of serum TSP-1 in diabetic patients without CAD and non-diabetic patients with CAD didn’t show significant difference. Plasma TSP-1 levels were higher in patients with DM than those in patients without DM (582.95 ± 55.70 ng/mL vs. 516.91 ± 64.56 ng/ml, P

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Generation of trans-arachidonic acid under nitrative stress is associated with upregulation of thromponsdin-1 in diabetic rats

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Chinese Medical Journal 2011年第12期124卷 1885-1889页

作者： XU Lin LIU Yu-min YAO Zhu-jun XUN Guo-liang YU Wen-juan LIU Kun SUN Xiao-dong QIU Yun-ping WU Hai-xiang XU Xun Department of Ophthalmology First People's Hospital ofShanghai Shanghai Jiao Tong University Shanghai 200080 China Shanghai Key Laboratory of Ocular Fundus Diseases Science andTechnology Commission of Shanghai Municipality Shanghai200080 China Instrumental Analysis Center Shanghai Jiao Tong University Shanghai200240 China School of PharmacyShanghai Jiao Tong University Shanghai200240 China State Key Laboratory of Bioorganic and Natural ProductsChemistry Shanghai Institute of Organic Chemistry ChineseAcademy of Sciences Shanghai 200032 China

Background Trans-arachidonic acids （TAAs）, newly discovered markers of nitrative stress and the major products of nitrogen dioxide （NO2）-mediated isomerization of arachidonic acid （AA）, represent a new mechanism of NO2-induced toxicity. It has been reported that TAAs were generated in oxygen-induced microvascular degeneration model and TAAs were also generated in a diabetic retinopathy （DR） model. In this study, we examined high glucose-induced nitrative stress damage and TAAs levels and explored the possible mechanisms for DR caused by reactive nitrogen species. Methods Diabetic rats were induced by intraperitoneal injection of streptozotocin （BTZ） at 60 mg/kg. Bovine retinal capillary endothelial cells （BRECs） were selectively cultured and incubated with normal or high glucose. The serum TAAs and AA in diabetic rats were measured by the gas chromatography and mass spectrometry （GC/MS） method. The ratio of peak area of TAAs to AA with selected ion of 79 was estimated by a group t-test. thrombospondin-1 （TSP-1） in the rat retinas and BRECs extracts were examined by Western blotting. The phosphorylation of extracellular signal-regulated kinase 1/2 （ERK1/2） protein was examined by Western blotting in BRECs incubated with high glucose. Results The TAAs to AA ratio （TAAs/AA） was significantly increased in the serum at 8, 12 and 16 weeks after BTZ injection （P 〈0.05）, with no noticeable change found at 2 or 4 weeks （P 〉0.05）. Expression of TSP-1 in the retina of diabetic rats was progressively elevated according to the duration of diabetes. TSP-1 expression was increased in BRECs incubated with high glucose all 48 hours. Moreover, high glucose also increased ERK1/2 expression, which peaked at 30 minutes and then decreased in the following 48 hours. Conclusion An elevation of TAAs/AA is associated with high glucose-induced nitrative stress, which probably involves upregulation of TSP-1 through activating ERK1/2.

关键词： trans-arachidonic acids nitrative stress diabetic retinopathy thrombospondin-1 nitrogen dioxide

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Inhibition of microvascular endothelial cell apoptosis by angiopoietin-1 and the involvement of cytochrome C

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Chinese Medical Journal 2006年第9期119卷 725-730页

作者： SHI Lian-guo ZHANG Guo-ping JIN Hui-ming Department of Physiology ＆ Pathophysiology Shanghai Medical College Fudan University Shanghai 200032 China

Background Angiopoietin-1 （Ang-1） is an endothelial-specific growth factor that can promote angiogenesis. Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murine- cerebral-derived microvascular endothelial cells （bEnd.3） induced by serum-free culture,we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C （Cyt C）. Methods The cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang-1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA. Results The apoptotic rate of bEnd.3 cells after stimulation with Ang-1 （100 ng/L） in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide （PI） and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3 cell apoptosis was strengthened with the increase in concentration （0-400 ng/ml）. Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1 significantly decreased CytC content in cytoplasm and increase that in mitochondria. Conclusions Ang-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed.

关键词： ANGIOGENESIS EXPRESSION thrombospondin-1

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Renal Protective Activity of Hsian-tsao Extracts in Diabetic Rats

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Biomedical and Environmental Sciences 2008年第3期21卷 222-227页

作者： MIN YANG ZHENG-PING XU CAI-JU XU JIA MENG GANG-QIANG DING XIAO-MING ZHANG YAN WENG Zhejiang University School of Medicine Hangzhou 310058 Zhejiang China Zhejiang Provincial Center for Disease Control and Prevention Hangzhou 310009 Zhejiang China

Objective To investigate the renal protective activity of Hsian-tsao Mesona procumbens Hemsl. water extracts in diabetic rats. Methods Thirty Sprague-dawley female rats were randomly divided into three groups （n=10 each）, ＂control group＂ with intraperitoneal saline injection, ＂diabetic group＂ with 60 mg of intraperitoneal streptozotocin injection per kg of body weight and ＂Hsian-tsao group＂ with intragastric administration of Hsian-tsao extraction everyday for 4 weeks after intraperitoneal streptozotocin injection. The body weight and blood sugar were measured before and after model induction in the three groups. thrombospondin-1 （TSP-1） expressions in the kidney were monitored by immunohistochemistry. Kidney ultrastructural changes were also analyzed by using transmission electron microscopy. Results Before diabetic model induction, there were no significant differences among the three groups in body weight and blood sugar. Four weeks after the induction of diabetes, the differences became statistically significant. Electron microscopy also revealed disruption of the foot processes of the podocytes and other damages in diabetic group. These damages were significantly less severe in Hsian-tsao group when compared with the diabetic group. TSP-1 expressions in the kidney were significantly increased in both the diabetic group and Hsian-tsao group, but it was relatively lower in Hsian-tsao group than in diabetic group. Conclusion Our results showed that Hsian-tsao treatment in the diabetic rats effectively prevented the pathological alterations in the kidney and decreased the TSP- 1 expression. It was suggested that Hsian-tsao had protective effect on the kidneys of the diabetic rats.

关键词： Rat Mesona procumbens Hemsl. Diabetic nephropathy thrombospondin-1

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