Jatonik polyherbal mixture induced rat liver MMPT pore opening in normal Wistar rat: In vitro and in vivo studies / 中草药·英文版

OBJECTIVE@#To assess acute toxicity, the in vitro and in vivo effects of methanol and ethyl acetate extracts (JME and JEE) of Jatonik polyherbal mixture on some mitochondria-related parameters and their effect on the activity of some liver enzymes.@*METHODS@#Acute toxicity of JME and JEE was determined using Lorke's method. In vitro and in vivo opening of the mitochondrial membrane permeability transition pore (MMPT pore) was spectrophotometrically assayed. Production of malondialdehyde (MDA) as an index of lipid peroxidation and the activity of mitochondrial ATPase was evaluated in vitro and in vivo and the effect of JME and JEE on the activity of liver enzymes such as alkaline phosphatase (ALP), aspartate and alanine aminotransferase (AST and ALT) and gamma-glutamyl transferase (GGT) was also investigated.@*RESULTS@#JME had an LD50 of 3 808 mg/kg b.w whereas JEE had an LD50 greater than 5 000 mg/kg b.w. of rats. After the rats have been fed with both extracts, a photomicrograph of a piece of liver tissue showed no apparent symptoms of toxicity. From the in vitro and in vivo studies, both extracts prompted intact mitochondria to open their MMPT pores. When compared to the control, lipid peroxide product release and ATPase activity were significantly increased (P < 0.05) in vitro and in vivo. The activities of AST, ALT, and GGT were all reduced at 50 mg/kg when treated with JME, but the activity of AST was considerably enhanced when treated with JEE (P < 0.05). The results revealed that both JME and JEE of the Jatonik polyherbal mixture had low toxicity, profound MMPTpore induction, and enhanced ATPase activity, but an increased MDA production.@*CONCLUSION@#Jatonik extracts may be a promising target for drug development in diseases where there is dysregulation of apoptosis, however, further studies are needed to better clarify the molecular mechanism involved in these phenomena.

Estudio Piloto Comparativo entre la Identificación Dactiloscópica y Poroscópica con Fines de Identificación Humana en el Perú / Comparative Pilot Study Between Dactyloscopy Identification and Poroscopy for Human Identification Purposes in Peru

La dactiloscopía o papiloscopía corresponde al estudio científico de las impresiones digitales, palmares y plantares, que tiene por finalidad la identificación infalible o indubitada del individuo. Existen tres niveles para identificar con mayor certeza nivel 1 (tipo o patrón dactilar), el nivel 2 (minucias o puntos característicos) y el nivel 3 (poroscopia y crestoscopia). Por ello, es necesario analizar las características de las impresiones dactilares directas y las huellas dactilares directas con la finalidad de verificar la presencia de puntos y poros característicos para mejorar el proceso de identificación humana. Se analizaron 80 muestras (54 mujeres y 26 hom- bres). A partir de ellos, se capturaron 800 impresiones y 800 huellas dactilares directas con tampón dactilar y polvo black. En huellas con tampón se identificaron 71.25 % y 1.25 % con 14 y 6 Puntos Característicos respectivamente y en grupos de poros el 84 % y 35 % para un grupo de 1 y grupos de 7 y 8 poros respectivamente. Con polvo black solo se identificaron Puntos Característico y no Poros. La cantidad de poros en hombres fue mayor igual a 10 (LR= 2.08) y en mujeres menor igual a 6 (LR= 1.93). Los grupos de poros fueron para hombres menores o iguales a 12 poros (LR= 1.04) y mayores o iguales a grupos de 13 poros (LR=1.28) para mujeres. Se consiguieron identificar grupos de poros con tampón dactilar pero no con polvos químicos lo que podría emplearse para implementar un protocolo para el uso del nivel 3 de identificación.

SUMMARY: Dactyloscopy or papiloscopy corresponds to the scientific study of digital, palmar and plantar impressions, whose purpose is the infallible or indubitable identification of a subject. There are three levels to identify with greater certainty level 1 (type or fingerprint pattern), level 2 (minutiae or characteristic points) and level 3 (poroscopy and crestoscopy). Therefore, it is necessary to analyze the characteristics of direct fingerprints and direct fingerprints in order to verify the presence of characteristic points and pores to improve the human identification process. 80 samples (54 women and 26 men) were analyzed. Of these, 800 impressions and 800 direct fingerprints with fingerprint buffer and black powder were captured. In footprints with buffer, 71.25 % and 1.25 % were identified with 14 and 6 Characteristic Points respectively and in groups of pores 84 % and 35 % for a group of 1 and groups of 7 and 8 pores respectively. With black powder, only Characteristic Points and no Pores were identified. The number of pores in men was greater than 10 (LR= 2.08) and in women less than 6 (LR= 1.93). The groups of pores were less than or equal to 12 pores (LR= 1.04) for men and greater than or equal to groups of 13 pores (LR=1.28) for women. It was possible to identify groups of pores with a fingerprint buffer but not with chemical powders, which could be used to implement a protocol for the use of level 3 identification.

Muscone inhibits opening of mPTP to alleviate OGD/R-induced injury of HT22 cells / 中国中药杂志

This study aims to investigate the mechanism of muscone in inhibiting the opening of mitochondrial permeability transition pore(mPTP) to alleviate the oxygen and glucose deprivation/reoxygenation(OGD/R)-induced injury of mouse hippocampal neurons(HT22). An in vitro model of HT22 cells injured by OGD/R was established. CCK-8 assay was employed to examine the viability of HT22 cells, fluorescence microscopy to measure the mitochondrial membrane potential, the content of reactive oxygen species(ROS), and the opening of mPTP in HT22 cells. Enzyme-linked immunosorbent assay was employed to determine the level of ATP and the content of cytochrome C(Cyt C) in mitochondria of HT22 cells. Flow cytometry was employed to determine the Ca~(2+) content and apoptosis of HT22 cells. The expression of Bcl-2(B-cell lymphoma-2) and Bcl-2-associated X protein(Bax) was measured by Western blot. Molecular docking and Western blot were employed to examine the binding between muscone and methyl ethyl ketone(MEK) after pronase hydrolysis of HT22 cell proteins. After the HT22 cells were treated with U0126, an inhibitor of MEK, the expression levels of MEK, p-ERK, and CypD were measured by Western blot. The results showed that compared with the OGD/R model group, muscone significantly increased the viability, mitochondrial ATP activity, and mitochondrial membrane potential, lowered the levels of ROS, Cyt C, and Ca~(2+), and reduced mPTP opening to inhibit the apoptosis of HT22 cells. In addition, muscone up-regulated the expression of MEK, p-ERK, and down-regulated that of CypD. Molecular docking showed strong binding activity between muscone and MEK. In conclusion, muscone inhibits the opening of mPTP to inhibit apoptosis, thus exerting a protective effect on OGD/R-injured HT22 cells, which is associated with the activation of MEK/ERK/CypD signaling pathway.

Advances on pathogenesis of acquired peritoneal ultrafiltration failure in peritoneal dialysis / 中华肾脏病杂志

Peritoneal ultrafiltration failure is a common reason for peritoneal dialysis (PD) withdrawal as well as mortality in PD patients. Based on the three-pore system, inter-cellular small pores and trans-cellular ultra-small pores (aquaporin-1) are mainly responsible for water transfer across the peritoneum. Both small and ultra-small pores-dependent water (free water) transport decline accompanied with time on PD, with more significant decrease in free water, resulting in peritoneal ultrafiltration failure. The reduction of free water transport is associated with fast peritoneal solute transfer, reduced crystalloid osmotic gradient due to increased interstitial glucose absorption, and declined osmotic conductance to glucose resulted from impaired aquaporin-1 function and peritoneal interstitial fibrosis. The decline of small pore-based water is mainly because of fast loss of crystalloid osmotic gradient, decrease of hydrostatic pressure mediated by peritoneal vasculopathy, as well as reduced absolute number of small pores. The current review discusses the advance on pathogenesis of acquired peritoneal ultrafiltration failure in long-term PD.

Discussion on Coronary Microcirculation Disorder after Myocardial Ischemia Reperfusion Based on “Collaterals-Sweat Pore Qi and Fluid” Theory / 中医杂志

Coronary microcirculation disorder after myocardial ischemia reperfusion （MIR） is a prominent problem in the treatment of coronary heart disease. According to the physiological commonality between “collaterals-sweat pore qi and fluid” and coronary microcirculation， and the evolution of the course of MIR， it is believed that “heart collateral stasis obstruction， sweat pore constraint and block” is the cause of coronary microcirculation disorder. The evolution of the pathogenesis can be divided into three periods. During the myocardial ischemia period， the pathogenesis is heart collaterals obstruction and sweat pores empty， while during the ischemia reperfusion period， it is internal formulation of deficiency wind， spasms of collaterals or slight heart collaterals obstruction； in the coronary microcirculation disorder period， sweat pores constraint and block， constraint transforming into heat， qi and fluid failing to diffuse are the pathogenesis. The corresponding treatment principle is assisting dredge with supplementation， and supplementing deficiency to dispel stasis； treating wind and blood simultaneously， and extinguishing wind to arrest convulsion； clearing heat and cooling blood， and diffusing qi and unblocking qi and fluid. Moreover， it is recommended to treat the heart and lungs simultaneously， and regulate the heart and liver at the same time.

Meridian Tropism and Compatibility of Chinese Medicine from Theory of Brain Sweat Pore / 中国实验方剂学杂志

Since LIU Hejian proposed the concept of sweat pore， the theory of sweat pore has experienced accelerated development. Especially with the advances in modern human anatomy and physiology， the microscopic anatomy of sweat pore begins to focus on the intercellular space， ion channels and other membranous space with channels， pores， doors， etc.， which exert the functions of exchanging fluid， information， and energy inside and outside blood vessels and discharging metabolic wastes so as to maintain the normal operation of organs. Therefore， sweat pore is the structural basis for the movement of Qi and the central link of Qi-fluid exchange in the body. The brain， as the house of original spirit， is in charge of the spirit of five Zang-organs. The brain sweat pore is pivotal for the circulation of Qi， blood， and fluid in the brain， and it is the structural basis for the normal physiological functions of the brain. The dysfunction of the brain sweat pore will cause the stagnation of Qi and the abnormal transport of blood and fluid. It will cause the abnormal exchange of Qi， liquid and other material and information， which fail to nourish the original spirt and cause the loss of vital activity， eventually leading to consciousness and emotion disorders. The treatment should focus on opening the brain sweat pore， smoothing the exchange of Qi and fluid inside and outside the pore， and restoring the Qi movement， so as to cure encephalopathy. At present， western medicine treatment of encephalopathy needs to solve the problem of drug efflux from the blood-brain barrier and improve the effective concentration of drugs into the brain. The structure and function of brain sweat pore is similar to those of the blood-brain barrier. The aromatic resuscitative medicines and wind-extinguishing medicines can open the brain sweat pore. When being combined with other medicines， they can lead the medicine to enter the brain to restore the Qi movement of the brain sweat pore and enhance the therapeutic effect. Liver-pacifying wind-extinguishing medicines， insect medicines， tonifying medicines， heat-clearing toxin-removing medicines， and damp-draining medicines can treat pathological factors such as wind， phlegm， stasis， deficiency， toxin， and dampness， respectively. These medicines， combined with the medicines with the tropism to brain meridians， can open the brain sweat pore and guide the medicine into the brain to enhance the effective concentration of the medicine， thereby enhancing the efficacy against encephalopathy.

Protective effect and mechanism of mesenchymal stem cell-derived extracellular vesicle on radiation-induced liver injury / 器官移植

Objective To evaluate the protective effect and the underlying mechanism of mesenchymal stem cell-derived extracellular vesicle (MSC-EV) on radiation-induced liver injury and liver cell line injury in mouse models. Methods C57BL/6 mice were randomly divided into the blank group, model group and MSC-EV treatment group (treatment group), with 9 mice in each group. AML12 cells were randomly divided into the control group, irradiation group and MSC-EV intervention group (intervention group). Animal and cell models with radiation-induced injury were established by one-time 15 Gy and 6 Gy X-ray irradiation, respectively. At 48 h after irradiation, liver tissues and serum samples of mice were collected and prepared for subsequent experiments. At 15 h post-irradiation, cell experiment was carried out. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and content of malondialdehyde (MDA) in liver tissues and cells were measured. The relative expression levels of interleukin (IL)-1β, IL-6, transforming growth factor (TGF)-β and CXC chemokine ligand (CXCL)10 messenger RNA (mRNA) were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). Liver tissues were prepared for hematoxylin-eosin (HE) staining to calculate liver pathological injury score. The apoptosis of liver tissues and cells was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and propidiumiodide (PI) staining, respectively. The expression levels of glutathione peroxidase 4 (GPX4) and ferroptosis suppressor protein 1 (FSP1) proteins were detected by Western blot. The production level of reactive oxygen species (ROS) was detected by dihydroethidine (DHE) staining. The fluorescence intensity of mitochondrial permeability transition pore (mPTP) was determined. Results Compared with the blank group, serum levels of AST and ALT were up-regulated, and the relative expression levels of IL-1β, TGF-β and CXCL10 mRNA in the mouse liver tissues were up-regulated, and MDA content was increased, liver injury score was elevated, cell apoptosis rate was increased, intracellular ROS level was elevated, and the relative expression levels of GPX4 and FSP1 proteins in the mouse liver tissues were down-regulated in the model group, and the differences were statistically significant (all P<0.05). Compared with the model group, serum levels of AST and ALT were decreased, and the relative expression levels of IL-1β, TGF-β and CXCL10 mRNA in the liver tissues of mice were down-regulated, MDA content was declined, liver injury score was declined, cell apoptosis rate was decreased, intracellular ROS level was decreased, and the relative expression levels of GPX4 and FSP1 proteins in the liver tissues of mice were up-regulated in the treatment group, and the differences were statistically significant (all P<0.05). Compared with the control group, cell apoptosis rate was increased, intracellular ROS level was elevated, the fluorescence intensity of mPTP was weakened, the relative expression levels of IL-1β, TGF-β and IL-6 mRNA were up-regulated, MDA content was increased, and the relative expression levels of GPX4 and FSP1 proteins were down-regulated in the irradiation group, and the differences were statistically significant (all P<0.05). Compared with the irradiation group, cell apoptosis rate was declined, intracellular ROS level was decreased, the fluorescence intensity of mPTP was strengthened, the relative expression levels of IL-1β, TGF-β and IL-6 mRNA were down-regulated, MDA content was decreased and the relative expression levels of GPX4 and FSP1 proteins were up-regulated in the intervention group, and the differences were statistically significant (all P<0.05). Conclusions MSC-EV may effectively alleviate radiation-induced liver injury by reducing ferroptosis of liver cells, enhancing antioxidant level and decreasing the production of lipid peroxide, thereby effectively alleviating radiation-induced liver injury.

Role of CypD-mediated mPTP in melatonin improving cognitive impairment induced by repeated exposure to sevoflurane / 中国药理学通报

Aim To investigate the involvement of cy-clophilin D ( CypD ) -mediated mitochondrial permeability transition pore ( mPTP) in the neuroprotective effects of melatonin on cognitive impairment induced by repeated exposure to sevoflurane in newborn animals. Methods Mice were randomly assigned into control group, sevoflurane ( Sevo) group, and melatonin pre-treatment + sevoflurane ( Sevo + Mel) group. JC-1 kit was used to assess the mitochondrial membrane potential ( MMP) ; Western blot analysis was used to evaluate the protein expressions of CypD, postsynaptic density protein 95 ( PSD95 ), and Synapsin-1; and behavioral test were employed to measure cognitive function. Results The MMP level in the Sevo group was significantly reduced compared to the control group (P < 0. 01 ), the expression of CypD increased (P <0. 05), whereas the expression of PSD95 and Synapsin-1 decreased ( P < 0. 01) . Furthermore, the new object recognition index and spatial memory ability both exhibited a significant decline (P < 0. 01, P < 0. 05). However, when compared to the Sevo group, Sevo + Mel group could raise the MMP level (P <0. 01), increase the expression of synaptic proteins ( P < 0. 05 ), decrease the expression of CypD (P <0. 01) and elevate the new object recognition index and the spatial memory capacity ( P < 0. 01 ). Conclusions Melatonin could ameliorate cognitive impairment induced by repeated exposure to sevoflurane in newborn mice, and the underlying mechanism may be attributed to the inhibition of mPTP mediated by CypD and the promotion of synaptic protein synthesis.

Isolation, characterization and modulatory potentials of β-stigmasterol, ergosterol and xylopic acid from Anchomanes difformis on mitochondrial permeability transition pore in vitro / 中草药·英文版

OBJECTIVE@#Secondary metabolites and polyphenolic compounds from medicinal plants have been demonstrated to have multiple biological functions with promising research and development prospects. This study examined the effect of β-stigmasterol (with ergosterol) and xylopic acid isolated from Anchomanes difformis on liver mitochondrial permeability transition pore (mPTP).@*METHODS@#The compounds were isolated by vacuum liquid chromatography. Mitochondrial swelling was assessed as changes in absorbance under succinate-energized conditions.@*RESULTS@#1H and 13C NMR spectroscopic elucidation of the isolates affirmed the presence of β-stigmasterol with ergosterol (1:0.3) and xylopic acid. The isolates reversed the increase in lipid peroxidation and inhibited the opening of mitochondrial permeability transition pores caused by calcium and glucose. Pharmacological inhibition of mPTP offers a promising therapeutic target for the treatment of mitochondrial-associated disorders.@*CONCLUSION@#Reduction in the activity of calcium ATPase and the expression of Caspase-3 and -9 were observed, suggesting that they could play a role in protecting physicochemical properties of membrane bilayers from free radical-induced severe cellular damage and be useful in the management of diseases where much apoptosis occurs.

Research progress in infection-associated virulence factors of Staphylococcus aureus / 中华临床感染病杂志

Staphylococcus aureus is an important pathogen of human infectious diseases, which can cause skin and soft tissue infections, endocarditis, necrotizing pneumonia, myelitis and other serious infectious diseases. With the use of antibiotics, Staphylococcus aureus is evolving to develop drug resistance; at the same time it produces a variety of virulence factors to attack the host. This article will review the recent advances of Staphylococcus aureus virulence factors associated with the three stages of infection and introduce the detection methods of virulence factors briefly.

Study on the mechanism of Wuzi-Yanzong-Wan-medicated serum interfering with the mitochondrial permeability transition pore in the GC-2 cell induced by atractyloside / 中国天然药物

Wuzi-Yanzong-Wan (WZYZW) is a classic prescription for male infertility. Our previous investigation has demonstrated that it can inhibit sperm apoptosis via affecting mitochondria, but the underlying mechanisms are unclear. The purpose of the present study was to explore the actions of WZYZW on mitochondrial permeability transition pore (mPTP) in mouse spermatocyte cell line (GC-2 cells) opened by atractyloside (ATR). At first, WZYZW-medicated serum was prepared from rats following oral administration of WZYZW for 7 days. GC-2 cells were divided into control group, model group, positive group, as well as 5%, 10%, 15% WZYZW-medicated serum group. Cyclosporine A (CsA) was used as a positive control. 50 μmol·L-1 ATR was added after drugs incubation. Cell viability was assessed using CCK-8. Apoptosis was detected using flow cytometry and TUNEL method. The opening of mPTP and mitochondrial membrane potential (MMP) were detected by Calcein AM and JC-1 fluorescent probe respectively. The mRNA and protein levels of voltage-dependent anion channel 1 (VDAC1), cyclophilin D (CypD), adenine nucleotide translocator (ANT), cytochrome C (Cyt C), caspase 3, 9 were detected by RT-PCR (real time quantity PCR) and Western blotting respectively. The results demonstrated that mPTP of GC-2 cells was opened after 24 hours of ATR treatment, resulting in decreased MMP and increased apoptosis. Pre-protection with WZYZ-medicated serum and CsA inhibited the opening of mPTP of GC-2 cells induced by ATR associated with increased MMP and decreased apoptosis. Moreover, the results of RT-qPCR and WB suggested that WZYZW-medicated serum could significantly reduce the mRNA and protein levels of VDAC1 and CypD, Caspase-3, 9 and CytC, as well as a increased ratio of Bcl/Bax. However, ANT was not significantly affected. Therefore, these findings indicated that WZYZW inhibited mitochondrial mediated apoptosis by attenuating the opening of mPTP in GC-2 cells. WZYZW-medicated serum inhibited the expressions of VDAC1 and CypD and increased the expression of Bcl-2, which affected the opening of mPTP and exerted protective and anti-apoptotic effects on GC-2 cell induced by ATR.

Effect of effective parts of Dracocephalum moldavica on the necroptosis of HBMECs after OGD/R injury / 药学学报

We investigated the ability of Dracocephalum moldavica (EPDM) flavonoids to protect human brain microvascular endothelial cells (HBMECs) from necroptosis induced by ischemia-reperfusion injury. To mimic the process of cerebral ischemia-reperfusion injury, a necroptosis model was established by treatment with the pan-cysteine aspartic acid protease (caspase) inhibitor Z-VAD-FMK combined with oxygen-glucose deprivation/re-oxygenation (OGD/R) injury using HBMECs. Cell proliferation and cytotoxicity (cell counting kit-8, CCK-8) was used to measure cell viability. A Hoechst33342/PI fluorescent double-staining method was exploited to determine the rate of cell necroptosis. A commercial kit was used to detect lactate dehydrogenase in the cell culture supernate. DCFH-DA probes, calcein AM and JC-1 probes were used to measure changes in ROS production, mitochondrial membrane permeability transformation pore (MPTP) opening and mitochondrial membrane potential (MMP), respectively. Enzyme-linked immunosorbent assay (ELISA) kits were chosen to detect the release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). Western blotting was used to detect necroptosis-related proteins. The results show that relative to control group, Z-VAD-FMK combined with OGD/R injury reduced cell viability, increased the necroptosis rate and the levels of LDH and ROS in HBMECs. The MPTP of the model group cells opened and the MMP reduced. TNF-α, IL-1β, and IL-6 levels were significantly elevated. Furthermore, the expression of receptor-interacting protein kinase 3 (RIP3) and mitochondrial phosphoglycerate mutase 5 (PGAM5) was significantly increased, accompanied by an increase of phosphorylated mixed-lineage kinase domain-like protein (p-MLKL)/MLKL. EPDM partially reversed the changes of the above-mentioned factors in HBMECs induced by Z-VAD-FMK plus OGD/R injury. These results indicate that EPDM may protect HBMECs from cerebral ischemia-reperfusion injury by inhibiting the RIP3/MLKL/PGAM5 pathway and MPTP opening to maintain mitochondrial function, thereby providing a scientific basis for the use of EPDM in the treatment of cerebral ischemia-related diseases.

Development and optimization of cosolvent-based blended Sertraline orodispersible films - A step to personalized medicine

Abstract Personalized medicine is gaining importance in pharmacotherapeutics as it allows tailoring the drug treatment to achieve the best patient response. Orodispersible film (ODF) is easy to formulate in hospitals, produces dose flexibility to suit an individual needs, particularly for patients suffer from swallowing issues or prohibited to take fluids. Sertraline Hydrochloride (SRT) was solubilized in several cosolvents, then different SRT ODFs based on five hydrophilic polymers namely; polyvinyl alcohol (PVA), hydroxylethyl cellulose (HEC), hydroxypropyl methylcellulose E5 LV (HPMC E5 LV), sodium alginate (NaAlg) and gelatin at two concentrations (2% and 4%) were developed and characterized. The outcomes were exposed to response surface analysis to obtain the desirability results to obtain the optimized formulation. Blended ODFs were developed from 4% PVA and 2% HEC in different blends and then potassium chloride (KCl) as a pore-forming agent was added to the best formulation to investigate its dissolution enhancement effect. F14 containing 4% PVA: 2% HEC 2:1 with 5% KCl showed best physicochemical properties of suitable pH (5.6), disintegration time (6 sec), good folding endurance which released 91 % SRT after 15 min. SRT ODF is an encouraging delivery system in the course of personalized medicine for the management of depression.

Asymmetric Opening of Mitochondrial Permeability Transition Pore in Mouse Brain Hemispheres: A Link to the Mitochondrial Calcium Uniporter Complex

Abstract The prolonged entry of large amounts of calcium into the mitochondria through the mitochondrial calcium uniporter complex (MCUC) may cause the permeability transition pore (mPTP) to open, which contributes to the pathogenesis of several diseases. Tissue-specific differences in mPTP opening due to variable expression of MCUC components may contribute to disease outcomes. We designed this study to determine differential mPTP opening in mitochondria isolated from different regions of mouse brain and kidney and to compare it with the expression of MCUC components. mPTP opening was measured using mitochondria isolated from the left/right brain hemispheres (LH/RH, respectively) and from kidney cortex/medulla, while the expression level of MCUC components was assessed from total cellular RNA. Interestingly, LH mitochondria showed less calcium-induced mPTP opening as compared to RH mitochondria at two different calcium concentrations. Conversely, mPTP opening was similar in the renal cortex and renal medulla mitochondria. However, the kidney mitochondria demonstrated bigger and faster mPTP opening as compared to the brain mitochondria. Furthermore, asymmetric mPTP opening in the LH and RH mitochondria was not associated with the expression of MCUC components. In brief, this study demonstrates thus far unreported asymmetric mPTP opening in mouse brain hemispheres that is not associated with the mRNA levels of MCUC components.

The pterygospinous foramen (Civinini) and the pterygoalar (crotaphitico-buccinatorius): laboratory findings / El foramen pterigoespinoso (Civinini) y el pterigoalar (crotafítico-buccinatorius): hallazgos de laboratorio

SUMMARY: The Pterygospinous foramen and pterygoalar foramen (crotaphitico-buccinatorius) are variant and atavic formations of the skull base and arise respectively from complete or incomplete idiopathic ossifications of the pterygospinous and pterygoalar ligaments. By proximity with areas of relevance for diagnosis and surgery, such as access pathways to the parapharyngeal and retropharyngeal spaces, it is necessary to be aware of these conditions due to the difficulties generated in surgical maneuvers and the promotion of compressive syndromes of mandibular nerve branches. This study was conducted on 45 samples of dry skulls and disarticulated sphenoid bones belonging to the collection of the Federal University of Juiz de Fora, Governador Valadares campus, Minas Gerais, Brazil. Our results indicated a total incidence of complete and incomplete pterygospinous and pterygoalar foramen (crotaphitico- buccinatorius) in 15, 5 % (7 skulls), with a higher incidence for the incomplete form of pterygospinous foramen (Civinini foramen) in 4 skulls (8.8 %), with 3 presenting unilaterally and 3 presenting bilaterally. The pterygoalar foramen (crotaphitico-buccinatorius or Hyrtl) was noted bilaterally in 1 of the skulls (2.2 %). The pterygospinous foramen and pterygoalar foramen are important findings, sometimes incidental, of an area of great anatomical expressiveness and pathological occurrences, besides the indispensable and unclear studies of phylogenetic order.

RESUMEN: El foramen pterigoespinoso y el foramen pterigoalar (crotafítico-buccinatorius) son formaciones variantes y atávicas de la base del cráneo y surgen respectivamente de osificaciones idiopáticas completas o incompletas, de los ligamentos pterigoespinoso y pterigoalar. Debido a la proximidad con áreas de relevancia para el diagnóstico y la cirugía, como las vías de acceso a los espacios parafaríngeo y retrofaríngeo, es necesario conocer estas condiciones por las dificultades que se generan en las maniobras quirúrgicas. Este estudio se realizó en 45 muestras de cráneos secos y huesos esfenoides desarticulados pertenecientes a la colección de la Universidad Federal de Juiz de Fora, campus Governador Valadares, Minas Gerais, Brasil. Nuestros resultados indicaron una incidencia total de foramen pterigoespinoso y pterigoalar completo e incompleto (crotafítico-buccinatorius) en el 15,5 % (7 cráneos), con una mayor incidencia de la forma incompleta de foramen pterigoespinoso (agujero de Civinini) en 4 cráneos (8,8 %), con 3 de presentación unilateral y 3 de presentación bilateral. El foramen pterigoalar (crotaphitico-buccinatorius o Hyrtl) se observó bilateralmente en 1 de los cráneos (2,2 %). El foramen pterigoespinoso y pterigoalar son hallazgos importantes, a veces incidentales, de un área de gran expresividad anatómica y ocurrencias patológicas, además de los estudios indispensables y poco claros de orden filogenético.

Applications of atomic force microscopy in immunology / 医学前沿

Cellular mechanics, a major regulating factor of cellular architecture and biological functions, responds to intrinsic stresses and extrinsic forces exerted by other cells and the extracellular matrix in the microenvironment. Cellular mechanics also acts as a fundamental mediator in complicated immune responses, such as cell migration, immune cell activation, and pathogen clearance. The principle of atomic force microscopy (AFM) and its three running modes are introduced for the mechanical characterization of living cells. The peak force tapping mode provides the most delicate and desirable virtues to collect high-resolution images of morphology and force curves. For a concrete description of AFM capabilities, three AFM applications are discussed. These applications include the dynamic progress of a neutrophil-extracellular-trap release by neutrophils, the immunological functions of macrophages, and the membrane pore formation mediated by perforin, streptolysin O, gasdermin D, or membrane attack complex.

The mechanism of tilianin against ischemia-reperfusion injury of H9c2 cells / 药学学报

Z-VAD-FMK was combined with hypoxia-reoxygenation (H/R) injury to establish a necroptosis model of H9c2 cells to mimic the pathological changes of myocardial ischemia reperfusion injury (MIRI) in vitro and to study the effect and mechanism of tilianin against myocardial ischemia-reperfusion injury. A cell counting kit-8 (CCK-8) was used to detect cell viability, and commercial kits were used to detect lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the cell culture supernatant. Hoechst 33342/PI immunofluorescence staining was used to detect cell death. DCFH-DA, BBcellProbeTMM61, and JC-1 probes were used to detect reactive oxygen species (ROS), mitochondrial permeability transition pore (mPTP), and mitochondrial membrane potential (MMP), respectively. An enzyme-linked immunosorbent assay (ELISA) method was used to detect the release of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). The results show that the cell viability, SOD activity, and MMP of the model group induced by H/R injury decreased, as compared with control group, but the necroptosis rate, LDH level, and ROS release increased significantly. Furthermore, mPTP of the model group cells opened, and TNF-α, IL-1β, and IL-6 levels were significantly higher. Molecular docking modeling showed that tilianin can bind to calmodulin-dependent protein kinase II (CaMKII), and Western blot results showed that compared with control group, the expression levels of p-CaMKII and phospho-mixed lineage kinase domain-like protein increased in the model group, and tilianin could decrease the expression level of these proteins. The above results indicate that tilianin can protect H9c2 cells by inhibiting the phosphorylation of CaMKⅡ at threonine 287, protecting mitochondrial function, and inhibiting the opening of mPTP to prevent necroptosis. This study has value for research on new methods to treat H/R injury.

Kangxian Yixin Prescription Interferes with Mitochondrial Permeability Transition Pore to Inhibit Cardiomyocyte Apoptosis / 中国实验方剂学杂志

Objective:To explore the mechanism of Kangxian Yixin prescription in regulating mitochondrial permeability transition pore（mPTP）and inhibiting cardiomyocyte apoptosis. Method:H9c2 cardiomyocytes were cultured routinely. After 8 h of starvation，the cells were divided into the normal group，model group，Kangxian Yixin prescription（0.25 g·L^-1） group，and cyclosporin A（CsA，10 μmol·L^-1） group and treated with the corresponding drugs for 24 h for follow-up experiments. The H9c2 cardiomyocyte hypertrophy model was induced by norepinephrine（NE），whose optimal concentration was determined by real-time polymerase chain reaction （Real-time PCR）. The degree of mPTP opening was detected by flow cytometry， followed by the measurement of mRNA and protein expression levels of apoptosis-related factors cyclophilin D（Cyp-D），cytochrome C（Cyt-C），and cysteine aspartate-specific protease-3（Caspase-3） after mPTP opening and the quantification of mitochondrial membrane potential. Result:When the concentration of NE was 200 μmol·L^-1， the mRNA expression levels of atrial natriuretic peptide（ANP） and brain natriuretic peptide（BNP） were the highest， implying that it was the optimal concentration to induce H9c2 cell hypertrophy. Compared with the normal group，the model group exhibited excessive opening of mPTP，weakened relative fluorescence intensity in mitochondria， decreased mitochondrial membrane potential（<italic>P</italic><0.05，<italic>P</italic><0.01），and elevated mRNA and protein expression of Cyp-D，Cyt-C，and Caspase-3（<italic>P</italic><0.05）. Compared with the model group，both Kangxian Yixin prescription and CsA inhibited mPTP opening，enhanced the relative fluorescence intensity of mitochondria， increased mitochondrial membrane potential（<italic>P</italic><0.05，<italic>P</italic><0.01），and lowered the mRNA and protein expression of Cyp-D，Cyt-C，and Caspase-3 （<italic>P</italic><0.05）. Conclusion:Kangxian Yixin prescription inhibits cardiomyocyte apoptosis possibly by regulating mPTP opening and inhibiting the expression of apoptosis-related factors Cyp-D，Cyt-C， and Caspase-3.

Shexiang Tongxin Dropping Pill () Reduces Coronary Microembolization in Rats via Regulation of Mitochondrial Permeability Transition Pore Opening and AKT-GSK3β Phosphorylation / 中国结合医学杂志

OBJECTIVE@#To investigate the protective effects of Shexiang Tongxin Dropping Pill (, STDP) following sodium laurate-induced coronary microembolization (CME) in rats.@*METHODS@#Forty rats were divided into 4 groups: the control (sham) group, CME group, low-dose STDP pretreatment group (20 mg·kg@*RESULTS@#The rats in the CME group showed a significant increase in the fibrinogen-like protein 2 expression level and mitochondrial dysfunction and a decrease in the expression level of antioxidant biomarkers (superoxide dismutase and catalase, P<0.01 for all). In contrast, the rats in the low- and high-dose STDP pretreatment groups showed a significant decrease in coronary microthrombi (P<0.05); moreover, STDP restored the antioxidant-related protein activities and mitochondrial function, inhibited mPTP opening, decreased AKT-Ser473 phosphorylation, and increased GSK3β-Ser9 phosphorylation (P<0.05 or P<0.01).@*CONCLUSION@#STDP may be useful for treatment of CME, possibly via regulation of mPTP opening and AKT/GSK3β phosphorylation.

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells / 中草药·英文版

Objective: Chrysophanol (Chry) displays potent anticancer activity in human cancer cells and animal models, but the cellular targets of Chry have not been fully defined. Herein, we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity. Methods: Human liver cancer cell line HepG2 was incubated. The cytotoxicity was evaluated by MTT assay. Mitochondria localization was evaluated by a confocal microscopy. Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer. The levels of ATP, mitochondrial superoxide anions, and GSH/GSSG were determined according to the assay kits. The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining, respectively. The expression of cyclophilin D (CyPD) was determined by immunoblot method, and the interaction between CyPD and Chry was analyzed by molecule docking procedure. Results: Chry itself mainly localized in mitochondria to cause mitochondrial dysfunction and cell death in HepG2 cells. As regard to the mechanism, cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore (mPTP) moderately suppressed cell death, indicating mPTP involved in the process of cell death. Further, Chry enhanced the protein expression of Cyclophilin D (CyPD) which is a molecular componentry and a modulator of mPTP, while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD. Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with −11.94 kal/mol of the binding affinity value. Besides, the mtDNA-deficient HepG