Protective role and mechanism of human umbilical cord mesenchymal stem cell-derived exosome in renal ischemia-reperfusion injury / 器官移植

Objective To investigate the protective effect of human umbilical cord mesenchymal stem cell-derived exosome (hucMSC-Exo) on renal ischemia-reperfusion injury (IRI), and to clarify the critical role and regulating mechanism of transient receptor potential canonical (TRPC) 6/poly adenosine-diphosphate-ribose polymerase (PARP) 1 signaling pathway during this process. Methods The hucMSC-Exo was extracted by ultracentrifugation, and identified by transmission electron microscope (TEM), nanoparticle tracing analysis and Western blot. SD rats were randomly divided into the sham operation group (group S), sham operation+TRPC6 inhibitor SKF96365 group (group SS), renal IRI group (group IRI), exosome treatment group (group EXO) and exosome +TRPC6 inhibitor SKF96365 group (group ES), with 6 rats in each group. Serum creatinine and blood urea nitrogen levels were detected. Pathological changes of renal tissues were observed by hematoxylin-eosin (HE) staining and Paller score was calculated. The expression levels of key molecules of necroptosis in rat renal tissues, including receptor-interacting protein kinase (RIPK)1, RIPK3 and mixed-lineage kinase domain-like protein (MLKL), TRPC6 and PARP1, were detected by Western blot. Results Typical saucer-like structure was observed under TEM. Nanoparticle tracing analysis showed that the average diameter of the extracted substance was 125.9 nm. Western blot revealed that the surface markers of CD9, CD63 and CD81 were positively expressed, confirmed that the extracted substance was exosome. Compared with group S, the serum creatinine and blood urea nitrogen levels were up-regulated, the pathological damage of renal tissues was worsened, Paller score was elevated, the relative expression levels of TRPC6 and PARP1 proteins were down-regulated, and the relative expression levels of RIPK1, RIPK3 and MLKL proteins were up-regulated in group IRI (all P < 0.05). Compared with group IRI, the serum creatinine and blood urea nitrogen levels were down-regulated, the pathological damage of renal tissues was mitigated, Paller score was decreased, the relative expression levels of TRPC6 and PARP1 proteins were up-regulated, and the relative expression levels of RIPK1, RIPK3 and MLKL proteins were down-regulated in group EXO (all P < 0.05). Compared with group EXO, the serum creatinine and blood urea nitrogen levels were up-regulated, the pathological damage of renal tissues was aggravated, Paller score was increased, the relative expression levels of TRPC6 and PARP1 proteins were down-regulated, and the relative expression levels of RIPK1, RIPK3 and MLKL proteins were up-regulated in group ES (all P < 0.05). Conclusions hucMSC-Exo may alleviate the necroptosis induced by renal IRI in rat models, which is related to the activation of TRPC6/PARP1 signaling pathway.

Inhibitory Effect of Paeonol on Vascular Smooth Muscle Cell Senescence by Reducing SIRT6/PARP1-mediated DNA Damage / 中国实验方剂学杂志

ObjectiveTo investigate whether the effects of paeonol （Pae） on angiotensin Ⅱ （AngⅡ）-induced senescence in vascular smooth muscle cells （VSMCs） were related to angiotensinogen of silencing regulatory information factor 6 （SIRT6）/adenosine diphosphate ribose polymerase 1 （PARP1） signaling pathway in VSMCs. MethodThe model of VSMC-stress aging induced by AngⅡ （100 nmol·L-1） was established. The rats were divided into normal group， model group， low， medium， and high-concentration Pae groups （30， 60， 120 μmol·L-1）. The positive rate of cell senescence was detected by SA-β-Gal staining， the ability of cell proliferation was detected by the cell counting kit-8 （CCK-8） method， the expression of SIRT6， PARP1， p16， p21， p53， proliferating cell nuclear antigen （PCNA）， deoxyribonucleic acid （DNA）-damaged protein γ-H2AX was detected by Western blot， and VSMC proliferation was detected by EdU staining. The silenced VSMCs were prepared by siRNA-SIRT6 transfection， and the protein expressions of SIRT6， PARP1， p16， and γ-H2AX in VSMCs silenced by SIRT6 were observed. ResultThe results of SA-β-Gal staining showed that the senescence positive rate of SA-β-Gal staining in the model group was higher than that in the normal group （P<0.01）， and the positive rate of SA-β-Gal staining in the Pae group was significantly lower than that in the model group （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the expression of PCNA， SIRT6， and PARP1 in the model group was down-regulated， and the expression of aging-related proteins p16， p21， p53， and γ-H2AX was up-regulated in the model group （P<0.05， P<0.01）. Compared with the model group， Pae promoted the protein expression of PCNA， SIRT6， and PARP1 and inhibited the protein expression of p16， p21， p53， and γ-H2AX in a dose-dependent manner （P<0.05， P<0.01）. The results of EdU staining showed that the number of EdU positive cells in the model group was lower than that in the normal group （P<0.01）， and the number of EdU positive cells in Pae groups was significantly higher than that in the model group （P<0.05， P<0.01）. After SIRT6 silencing， the effects of Pae on promoting SIRT6 and PARP1 and inhibiting P16 were reversed （P<0.05， P<0.01）. In addition， the addition of SIRT6 inhibitor （IN-1） promoted the occurrence of cell senescence induced by AngⅡ （P<0.05， P<0.01）. ConclusionPae can effectively inhibit the aging of VSMCs， and its mechanism may be related to the regulation of SIRT6/PARP1 signal pathway.

Effect of poly adenosine diphosphate-ribose polymerase-1 inhibitor fluzoparib on the proliferation and migration of pancreatic cancer cell PANC1 / 中华胰腺病杂志

Objective:To investigate the effects of poly adenosine diphosphate ribose polymerase-1(PARP-1) inhibitor fluzoparib on proliferation, apoptosis and migration of pancreatic cancer PANC1 cells.Methods:PANC1 cells cultured in conventional culture medium were used as control group, and PANC1 cells cultured in the medium containing fluzoparib were used as fluzoparib group. The effects of fluzoparib with different concentrations on the proliferation of PANC1 cells were detected by CCK8 method, and the half inhibitory concentration (IC 50) of fluzoparib on PANC1 cells was calculated. The effect of fluzoparib on apoptosis and cell cycle of PANC1 cells was detected by flow cytometry, and the migration ability of PANC1 cells was detected by cell scratch test and Transwell chamber. Results:Compared with control group, with the increase of fluzoparib concentration and the prolongation of the action time, the cell proliferation activity of PANC1 in fluzoparib group was significantly decreased, and the differences were statistically significant (all P values <0.05). IC 50 of fluzoparib on PANC1 cells cultured for 24 h was 0.03 mmol/L. After 24 h culture, the IC 50 apoptosis rate of fluzoparib group was (32.19±2.48)%, and the apoptosis rate of control group was (21.99±6.30)%. The former was greatly higher than the latter, and the difference was statistically significant ( P<0.05). The proportion of cells in G 2/M phase was (16.28±0.62)% in the fluzoparib group and (11.64±0.88)% in the control group, and the difference between the two groups was statistically significant ( P<0.05). The migration rates of PANC1 cells in IC 50 fluzoparib group in 12 h and 24 h culture were (2.59±1.46)% and (19.76±7.84)%; and those in control group were (27.08±2.17)% and (45.92±3.61)%, respectively. The number of transmembrane cells was (348±19) cells/10 visual field in the fluzoparib group and (587±14) cells/10 visual field in the control group. The migration ability of PANC1 cells in fluzoparib group was significantly lower than that in control group ( P<0.05). Conclusions:Fluzoparib can inhibit the proliferation and migration of PANC1 cells and promote the apoptosis of PANC1 in vitro, which may be an effective drug for the treatment of pancreatic cancer.

Targeting whole body metabolism and mitochondrial bioenergetics in the drug development for Alzheimer's disease

Aging is by far the most prominent risk factor for Alzheimer's disease (AD), and both aging and AD are associated with apparent metabolic alterations. As developing effective therapeutic interventions to treat AD is clearly in urgent need, the impact of modulating whole-body and intracellular metabolism in preclinical models and in human patients, on disease pathogenesis, have been explored. There is also an increasing awareness of differential risk and potential targeting strategies related to biological sex, microbiome, and circadian regulation. As a major part of intracellular metabolism, mitochondrial bioenergetics, mitochondrial quality-control mechanisms, and mitochondria-linked inflammatory responses have been considered for AD therapeutic interventions. This review summarizes and highlights these efforts.

Mitochondrial complex I as a therapeutic target for Alzheimer's disease

Alzheimer's disease (AD), the most prominent form of dementia in the elderly, has no cure. Strategies focused on the reduction of amyloid beta or hyperphosphorylated Tau protein have largely failed in clinical trials. Novel therapeutic targets and strategies are urgently needed. Emerging data suggest that in response to environmental stress, mitochondria initiate an integrated stress response (ISR) shown to be beneficial for healthy aging and neuroprotection. Here, we review data that implicate mitochondrial electron transport complexes involved in oxidative phosphorylation as a hub for small molecule-targeted therapeutics that could induce beneficial mitochondrial ISR. Specifically, partial inhibition of mitochondrial complex I has been exploited as a novel strategy for multiple human conditions, including AD, with several small molecules being tested in clinical trials. We discuss current understanding of the molecular mechanisms involved in this counterintuitive approach. Since this strategy has also been shown to enhance health and life span, the development of safe and efficacious complex I inhibitors could promote healthy aging, delaying the onset of age-related neurodegenerative diseases.

Progress of drug resistance mechanism of poly adenosine diphosphate ribose polymerase inhibitors and the combined medication in patients with ovarian cancer / 肿瘤研究与临床

Ovarian cancer seriously threats women's health. The emergence of poly adenosine diphosphate ribose polymerase inhibitor (PARPi) has broken the barrier for ovarian cancer treatment. PARPi has been widely used and along with it comes the problem of drug resistance. Fully understanding the drug resistance mechanism of PARPi is expected to be an important way to reverse PARPi resistance. The combination of PARPi and other drugs for ovarian cancer may expand the benefits of patients using PARPi. This article reviews the drug resistance mechanism of PARPi and combined medication.

Research progress of NF-κB signaling pathway in kidney ischemia-reperfusion injury / 器官移植

Kidney ischemia-reperfusion injury (IRI) is the major cause of poor prognosis after kidney transplantation and partial nephrectomy. Besides, it is also a critical pathophysiological process of acute kidney injury. Consequently, the prevention and treatment of kidney IRI are of significance to improve clinical prognosis of recipients undergoing kidney transplantation. However, the mechanism underlying IRI is complex, and the exact mechanism remains elusive. Inflammation, as one of the main pathogenesis of IRI, plays a significant role in IRI-induced kidney injury. Nuclear factor (NF)-κB, as a rapid response transcription factor, has been proven to be involved in the regulation of inflammation during kidney IRI. Therefore, in this article, the structure of NF-κB, the activation pattern of NF-κB signaling pathway, the regulatory mechanisms of NF-κB upstream and downstream signaling pathways in kidney IRI were reviewed, and the role of NF-κB signaling pathway in kidney IRI was investigated, aiming to provide novel clinical ideas for the prevention and treatment of kidney IRI.

Progress of hereditary breast cancer with breast cancer susceptibility gene 1/2 mutations / 肿瘤研究与临床

Hereditary breast cancer refers to malignant tumors caused by pathogenic germline mutations of breast cancer susceptibility genes (BRCA). At present, it is believed that BRCA1/2 genes are most closely related to the development of hereditary breast cancer. Mutation will lead to loss of normal function, instability of genome, and then lead to tumorigenesis. Especially for those with germline mutations, not only the risk of breast cancer will be greatly increased, but also the probability of ovarian cancer and other cancers will be increased. With the emergence and clinical application of poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitors, BRCA1/2 genes have been regarded as new targets for the treatment of breast cancer. This article reviews the latest research of breast cancer with BRCA1/2 gene mutations.

Ginsenosides in

Ginsenosides are a series of glycosylated triterpenoids which belong to protopanaxadiol (PPD)-, protopanaxatriol (PPT)-, ocotillol (OCT)- and oleanane (OA)-type saponins known as active compounds of

The disruption of protein-protein interactions with co-chaperones and client substrates as a strategy towards Hsp90 inhibition

The 90-kiloDalton (kD) heat shock protein (Hsp90) is a ubiquitous, ATP-dependent molecular chaperone whose primary function is to ensure the proper folding of several hundred client protein substrates. Because many of these clients are overexpressed or become mutated during cancer progression, Hsp90 inhibition has been pursued as a potential strategy for cancer as one can target multiple oncoproteins and signaling pathways simultaneously. The first discovered Hsp90 inhibitors, geldanamycin and radicicol, function by competitively binding to Hsp90's N-terminal binding site and inhibiting its ATPase activity. However, most of these N-terminal inhibitors exhibited detrimental activities during clinical evaluation due to induction of the pro-survival heat shock response as well as poor selectivity amongst the four isoforms. Consequently, alternative approaches to Hsp90 inhibition have been pursued and include C-terminal inhibition, isoform-selective inhibition, and the disruption of Hsp90 protein-protein interactions. Since the Hsp90 protein folding cycle requires the assembly of Hsp90 into a large heteroprotein complex, along with various co-chaperones and immunophilins, the development of small molecules that prevent assembly of the complex offers an alternative method of Hsp90 inhibition.

Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion / 浙江大学学报·医学版

To investigate the effects of salt-inducible kinase 2 (SIK2) on energy metabolism in rats with cerebral ischemia-reperfusion. Adult SD male rats were divided into 5 groups: sham group, ischemia group, reperfusion group, adenovirus no-load group, and SIK2 overexpression group with 5 animals in each group. The middle cerebral artery occlusion (MCAO) was induced with the modified Zea-Longa line thrombus method to establish the cerebral ischemia reperfusion model. Eight days before the MCAO, SIK2 overexpression was induced by injecting 7 μL adenovirus in the right ventricle, then MCAO was performed for followed by reperfusion HE staining was used to observe the pathological changes of cerebral tissue in rats; TTC staining was used to observe the volume of cerebral infarct. The levels of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) in rat brain tissue were detected by ELISA; the levels of SIK2 and hypoxia-inducible factor 1α (HIF-1α) in the rat brain tissues were detected by RT-qPCR and Western blotting. Compared with the sham group, SIK2 level was decreased in the ischemia group, and it was further declined in the reperfusion group (<0.05). Compared with the sham group and ischemic group, the pathological injury in reperfusion group were more severe, and the infarct size was larger; compared with the reperfusion group and adenovirus no-load group, the pathological injury of the SIK2 overexpression group was milder, and the infarct size is less. Compared with the sharn group, HIF-1α was increased in both ischemia group and reperfusion group, especially in ischemia group (all <0.05); HIF-1α level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05). ATP level in ischemia group and reperfusion group was lower than that in the sham group, and the reperfusion group decreased more significantly than the ischemia group (<0.05); ADP content was increased in the ischemia and reperfusion group, and the ADP content in reperfusion group was significantly higher than that in the ischemia group (<0.05). ATP level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05), and ADP was decreased in the SIK2 overexpression group (all <0.05). SIK2 can up-regulate the ATP level and down-regulate the ADP level in rat brain tissue and alleviate cerebral ischemia-reperfusion injury by increase the level of HIF-1α.

ADP-ribosylhydrolases: from DNA damage repair to COVID-19 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Adenosine diphosphate (ADP)-ribosylation is a unique post-translational modification that regulates many biological processes, such as DNA damage repair. During DNA repair, ADP-ribosylation needs to be reversed by ADP-ribosylhydrolases. A group of ADP-ribosylhydrolases have a catalytic domain, namely the macrodomain, which is conserved in evolution from prokaryotes to humans. Not all macrodomains remove ADP-ribosylation. One set of macrodomains loses enzymatic activity and only binds to ADP-ribose (ADPR). Here, we summarize the biological functions of these macrodomains in DNA damage repair and compare the structure of enzymatically active and inactive macrodomains. Moreover, small molecular inhibitors have been developed that target macrodomains to suppress DNA damage repair and tumor growth. Macrodomain proteins are also expressed in pathogens, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, these domains may not be directly involved in DNA damage repair in the hosts or pathogens. Instead, they play key roles in pathogen replication. Thus, by targeting macrodomains it may be possible to treat pathogen-induced diseases, such as coronavirus disease 2019 (COVID-19).

Spatial-resolved metabolomics reveals tissue-specific metabolic reprogramming in diabetic nephropathy by using mass spectrometry imaging

Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy (DN) is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies. In the present study, a spatial-resolved metabolomics approach based on air flow-assisted desorption electrospray ionization (AFADESI) and matrix-assisted laser desorption ionization (MALDI) integrated mass spectrometry imaging (MSI) was proposed to investigate tissue-specific metabolic alterations in the kidneys of high-fat diet-fed and streptozotocin (STZ)-treated DN rats and the therapeutic effect of astragaloside IV, a potential anti-diabetic drug, against DN. As a result, a wide range of functional metabolites including sugars, amino acids, nucleotides and their derivatives, fatty acids, phospholipids, sphingolipids, glycerides, carnitine and its derivatives, vitamins, peptides, and metal ions associated with DN were identified and their unique distribution patterns in the rat kidney were visualized with high chemical specificity and high spatial resolution. These region-specific metabolic disturbances were ameliorated by repeated oral administration of astragaloside IV (100 mg/kg) for 12 weeks. This study provided more comprehensive and detailed information about the tissue-specific metabolic reprogramming and molecular pathological signature in the kidney of diabetic rats. These findings highlighted the promising potential of AFADESI and MALDI integrated MSI based metabolomics approach for application in metabolic kidney diseases.

Quality evaluation of Xiaojin Pills based on antiplatelet aggregation biological potency assay / 中草药

Objective: To establish a biological potency assay for Xiaojin Pills against platelet aggregation in vitro, evaluate the quality consistency of Xiaojin Pills, and screen traditioanal Chinese medicines which play the role of promoting blood circulation in Xiaojin Pills. Methods: Xiaojin Pills and ten Chinese medicines [artificial musk, Momordica cochinchinensis, Aconitum kusnezoffii, Liquidambar formosana, Boswellia carterii, Commiphora myrrha, Faeces Trogopterori, Angelica sinensis, Pheretima aspergillum, Fragrant Ink] in its formula were extracted by ultrasound in 40% methanol. The antiplatelet aggregation rate of the extract was measured by platelet aggregation meter. The platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from abdominal aorta of rats. The platelet aggregation was induced by adenosine diphosphate (ADP). With sodium ferulate as a standard reference material, the biological potency of antiplatelet aggregation of Xiaojin Pills was calculated by the simplified probit principle. Results: The results showed that the biological potency of Xiaojin Pills was between 0.598 and 1.338 U/mg among different manufacturers and batches. In Xiaojin Pills group, Pheretima, Faeces Trogopterori, and Momordicae Semen had stronger inhibitory effects on platelet aggregation with inhibition rates of 70.87%, 31.83% and 67.52%, respectively. Conclusion: The quality consistency of Xiaojin Pills from different manufacturers and batches is poor, and Pheretima, Faeces Trogopterori, and Momordicae Semen may be the key drugs for Xiaojin Pills to play the role of promoting blood circulation.

Discussion on medicinal attribution of Descurainia sophia and its nature and flavor splitting components based on typical energy metabolism of heat syndrome animal models / 中草药

Objective: The relevant indicators of energy metabolism in rats with heat syndrome were tested to verify the medicinal properties of Descurainia sophia and its nature and flavor splitting components, in order to explain the cold and heat properties of D. sophia and its splitting components. Methods: Healthy male Sprague-Dawley rats were randomly divided into normal control group (NC), model group (M), water extract of gardenia (DS), flavonoid glycosides composition of D. sophia (FG), flavonoid aglycone composition of D. sophia (FA), oligosaccharide resolution component group (Oli), gardenia polysaccharide decomposition component group (Pol), and D. sophia fatty oil component separation group (FO). The model of heat syndrome was established by intragastric administration of Euthyrox (120 mg/kg). After 15 days of continuous administration, blood was taken from the abdominal aorta and the liver and heart were taken to detect the indicators related to the energy metabolism of the substance. The enzyme expression of glucokinase (GCK) and fructose phosphokinase (PFK-1), phosphoglycerate kinase (PGK), pyruvate kinase (PK), pyruvate dehydrogenase (PDH), acetyl-CoA, citrate synthase (CS), isocitrate dehydrogenase (ICD), alpha-ketoglutarate dehydrogenase (alpha-KGDHC), fumarate (FUM), glycogen phosphorylase (PYGL), glycogen synthase kinase-3 (GSK-3), adipose triglyceride lipase (ATGL), cytochrome C reductase (CCR), cytochrome C oxidase (COX), ATP synthase (ATPs), adenylate kinase (ADK), Na+ K,+-ATPase and the content of adenosine triphosphate (ATP), adenosine diphosphate (ADP), nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH) were determined. Results: Compared with NC group, the weight of M group rats was decreased significantly (P < 0.01). Compared with M group, the body weights of rats in DS, FG, FA, Pol and tFO groups were significantly or highly significantly increased (P < 0.05, 0.01). Compared with NC group, the number of spontaneous activities in M group was increased significantly (P < 0.01) within 5 min, and the number of locomotor activities of each group within 5 min after administration was significantly decreased (P < 0.01). Compared with NC group, the levels of GCK, PFK-1, PK, PGK, PDH, acetyl-CoA, CS, ICD, α-KGDHC, FUM, PYGL, GSK-3, ATGL, ATP, CCR, COX, ATPs, ADK, Na+ K,+-ATP, NADH expression or content in M group of rats were significantly increased (P < 0.05, 0.01), the content of ADP, NAD+ and NAD+/NADH was decreased (P < 0.05, 0.01). After administration, the expression level of material energy metabolism of the rats in each group was significantly or extremely significantly reduced compared with the M group (P < 0.05, 0.01). Conclusion: D. sophia can improve the state of the energy-metabolism related indicators of the rats in the heat syndrome model group. It is verified that D. sophia nature and flavor splitting components show the cold (cool) properties. From the side, it reflects the mechanism of "treating heat with cold drug" is related with the substance energy metabolism.

Characterization of the distinct mechanism of agonist-induced canine platelet activation

Platelet activation has a major role in hemostasis and thrombosis. Various agonists including adenosine diphosphate (ADP) and thrombin interact with G protein-coupled receptors (GPCRs) which transduce signals through various G proteins. Recent studies have elucidated the role of GPCRs and their corresponding G proteins in the regulation of events involved in platelet activation. However, agonist-induced platelet activation in companion animals has not been elucidated. This study was designed to characterize the platelet response to various agonists in dog platelets. We found that 2-methylthio-ADP-induced dog platelet aggregation was blocked in the presence of either P2Y₁ receptor antagonist MRS2179 or P2Y₁₂ receptor antagonist AR-C69931MX, suggesting that co-activation of both the P2Y₁ and P2Y₁₂ receptors is required for ADP-induced platelet aggregation. Thrombin-induced dog platelet aggregation was inhibited in the presence of either AR-C69931MX or the PKC inhibitor GF109203X, suggesting that thrombin requires secreted ADP to induce platelet aggregation in dog platelets. In addition, thrombin-mediated Akt phosphorylation was inhibited in the presence of GF109203X or AR-C69931MX, indicating that thrombin causes Gi stimulation through the P2Y₁₂ receptor by secreted ADP in dog platelets. Unlike human and murine platelets, protease-activated receptor 4 (PAR4)-activating peptide AYPGKF failed to cause dog platelet aggregation. Moreover, PAR1-activating peptide SFLLRN or co-stimulation of SFLLRN and AYPGKF failed to induce dog platelet aggregation. We conclude that ADP induces platelet aggregation through the P2Y₁ and P2Y₁₂ receptors in dogs. Unlike human and murine platelets, selective activation of the PAR4 receptor may be insufficient to cause platelet aggregation in dog platelets.

Expression differences of poly adenosine diphosphate ribose polymerase 1 in the human intracranial ruptured and unruptured aneurysm walls / 中国脑血管病杂志

Objective To investigate the expression differences of poly adenosine diphosphate ribose polymerase 1 (PARP-1) in the human intracranial ruptured and unruptured aneurysm walls. Methods Consecutive patients with intracranial aneurysm treated with craniotomy clipping surgery in Drum Tower Hospital, Nanjing University School of Medicine (n = 1) and General Hospital of Eastern War Zone (n = 12) from January 2014 to September 2018 were enrolled retrospectively. Seven of them were ruptured aneurysms and 6 were unruptured aneurysms. The expression of PARP-1 in aneurysm walls was detected by immunohistochemistry. Results Immunohistochemistry staining showed that the positive expression of PARP-1 was brown-yellow and mainly located in nuclei; in the unruptured aneurysm walls,the number of PARP-1 positive cells was small, the distribution was sparse,and it expressed in the whole layer of the aneurysm wall;in the ruptured aneurysm walls,PARP-1 positive cells were densely distributed in the outer membrane of the vessel walls. Statistical analysis showed that the positive expression of PARP-1 in the ruptured aneurysm walls was higher than that in the unruptured aneurysm walls. The difference was statistically significant(160 ± 19 vs. 56 ± 14,1 = 4. 291, P < 0. 01) . Conclusion The expression of PARP-1 in the ruptured aneurysm walls is significantly higher than that in the unruptured aneurysms,which may play a role in the rupture of intracranial aneurysms.

Effect of electroacupuncture pretreatment on adenine nucleotides in myocardial tissues of rats with myocardial ischemia-reperfusion injury detected by high performance liquid chromatography (HPLC) / 针灸推拿医学（英文版）

Objective:To observe the effect of electroacupuncture (EA) pretreatment on adenine nucleotides in the myocardial tissues of the myocardial ischemia-reperfusion injury (MIRI) rats, and to explore the mechanism of EA pretreatment on myocardial prevention and protection in MIRI rats. Methods:Forty SPF male Sprague-Dawley (SD) rats were randomly divided into 5 groups: a blank group, a sham operation group, a model group, an EA at Neiguan (PC 6) group and an EA at Hegu (LI 4) group, with 8 rats in each group. Rats in the blank group only received binding to the rat plate, 30 min/time, once a day for 7 d; on the 7th day, rats in the sham operation group were subjected to threading for 40 min at the left anterior descending coronary artery without ligation, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day, rats in the model group were subjected to threading at the left anterior descending coronary artery with ligation, for 40 min before the blood flow was restored, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day of pretreatment with EA at Neiguan (PC 6) or Hegu (LI 4) for 30 min per day (once a day for 7 d), rats in the EA at Neiguan (PC 6) group and EA at Hegu (LI 4) group were subjected to modeling and sample collection same as in the model group. The left ventricular myocardium of the lower left anterior descending coronary artery was collected from rats in all 5 groups. Hematoxylin-eosin (HE) staining and transmission electron microscope (TEM) were used to observe the changes in myocardial pathological morphology. The change in the adenine nucleotide level of myocardial tissue was measured by high performance liquid chromatography (HPLC). Results:The HE staining and ultrastructure showed that the myocardial injury was severer in the model group compared with the sham operation group. Compared with the model group, the myocardial injury in the EA at Neiguan (PC 6) and the EA at Hegu (LI 4) groups was mild or hardly any. The adenine nucleotide levels in the sham operation group and the model group were all decreased compared with the blank group (allP<0.05); compared with the sham operation group, the adenine nucleotide level of the model group was also decreased, but the difference was not statistically significant (P>0.05); compared with the model group, the adenine nucleotide level in the EA at Neiguan (PC 6) group was increased (P<0.05), and the adenine nucleotide level in the EA at Hegu (LI 4) group was significantly increased (P<0.01). The adenine nucleotide level in the EA at Hegu (LI 4) group was higher than that in the EA at Neiguan (PC 6) group, but the difference was not statistically significant (P>0.05). Compared with the EA at Neiguan (PC 6) group, the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in the EA at Hegu (LI 4) group were significantly increased (allP<0.01). Conclusion:Both EA at Neiguan (PC 6) and Hegu (LI 4) can alleviate the pathological damage to myocardium in MIRI rats, and increase the adenine nucleotide level in myocardial tissues, and thus protect MIRI rats. EA at Hegu (LI 4) has a better protective effect than Neiguan (PC 6).

Association between poly (ADP-ribose) polymerase-1 polymorphism and obesity and non-alcoholic fatty liver disease among children and adolescents / 中国学校卫生

Objective@#To study the relationship between rs1136410 polymorphism in poly (ADP-ribose) polymerase-1 gene (PARP-1) with obesity and non-alcoholic fatty liver disease (NAFLD) in children and adolescents, and to provide a reference for demonstrating the mechanism of obesity and NAFLD of children and adolescents and making the early prevention strategies.@*Methods@#In total, 2 030 children and adolescents aged 7-18 years old were recruited. Anthropometric measurements were performed. Liver B-ultrasound detection were performed in a subgroup. The matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TDF MS) was used for genotyping the rs1136410 polymorphism. Logistic regression was used to analyze the relationship between rs1136410 polymorphism and obesity or NAFLD in children and adolescents.@*Results@#Totally, 1 325 subjects were in non-obese group and 705 subjects were in obese group. The frequency of G allele and A allele were 43.86% and 56.14%. After adjusting for gender, age, and study population, the risk of obesity increased by 1.17 times for each additional A allele of rs1136410 polymorphism(OR=1.17, 95%CI=1.02-1.33, P=0.03). And the risk of NAFLD increased by 1.43 times for each additional A allele of rs1136410 polymorphism(OR=1.43, 95%CI=1.11-1.85, P=0.01). After further adjustment for BMI, rs1136410 polymorphism was not associated with NAFLD(P=0.70).@*Conclusion@#The rs1136410 polymorphism in PARP-1 gene is associated with obesity and NAFLD in children and adolescents. The effect of the gene polymorphism on NAFLD is mediated by BMI.

Analysis of the efficacy of poly ADP?ribose polymerases inhibitor combined with chemotherapeutics in the treatment of triple negative breast cancer / 中国综合临床

Objective To investigate the effects of poly adenosine diphosphate ribose polymerase (PARP) inhibitor AG014699 on the proliferation of triple negative breast cancer (TNBC) cell line MDA?MB?231.Methods Cell proliferation and cytotoxicity test kit ( CCK?8) was used to detect the proliferation of MDA?MB?231 cells in different concentrations of AG014699 (0.1,1.0,10.0,20.0 and 40.0 mmol／L), DTX (10－9,10－8,10－7,10－6 and 10－5 mol／L) and CBP (10－6,10－5,10－4 and 10－3 mol／L).Flow cytometry was used to detect cell apoptosis and cell cycle distribution.Results The effects of AG01469 at different concentrations (0.1,1.0,10.0,20.0 and 40.0 μmol／L) on proliferation activity of MDA?MB?231 cells were (94.83 ± 3.93)%, ( 79.42 ± 5.52)%, ( 63.75 ± 4.34)%, ( 38.97 ± 8.42)%, ( 29.70 ± 3.35 )%, with statistically significant differences (F＝75.54,P＜0.01,different concentrations pairwise comparison: all P ＜0.05). The efficacy of AG014699 in combination with DTX at different concentrations (( 69.77 ±17.94)%,(58.34± 2.59)%,( 52.81 ± 2.01)%, ( 41.23 ± 3.38)%, ( 24.82 ± 0.73)%) was compared with that of single DTX (( 81.24 ± 11.91)%, ( 85.74 ± 3.10)%, ( 72.74 ± 4.66)%, ( 55.18 ± 3.19)%, (45.95±3.82)%).The differences were statistically significant (t values were －0.923,－11.748,－6.802,－5.199,－9.410,respectively,with P＞0.05 at 10－9 concentration and P＜0.01 at all other concentrations ). The efficacy of AG014699 combined with CBP ((78.33± 2.89)%,( 60.44± 1.95)%,( 50.55± 3.07)%, (12.07± 1.63)%) and single CBP (( 90.00 ± 6.18)%, ( 87.87 ± 2.30)%,( 76.82 ± 3.37)%,( 40.71 ±1.68)%) was compared,and the cell activity was significantly reduced,indicating statistically significant differences ( t values were －1.935,－15.756,－9.981,－21.192, respectively, and P＞0.05 at 10－6 concentration,P＜0.05 at all the other concentrations ).The q value was＞1.15 when AG014699 was combined with 10－3 mmol／L CBP, which showed synergistic effect.When combined with other effective concentrations of DTX or CBP,the q value was between 0.85 and 1.15,showing additive effect.Conclusion PARP inhibitor AG014699 assisted DTX or CBP can inhibit the proliferation of TNBC cell line MDA?MB?231.By means of simple addition or systematic effect,it can inhibit the triple negative breast cancer.