Highly Aligned Ternary Nanofiber Matrices Loaded with MXene Expedite Regeneration of Volumetric Muscle Loss

Current therapeutic approaches for volumetric muscle loss(VML)face challenges due to limited graft availability and insufficient bioactivities.To overcome these limitations,tissue-engineered scaffolds have emerged as a promising alternative.In this study,we developed aligned ternary nanofibrous matrices comprised of poly(lactide-co-ε-caprolactone)integrated with collagen and Ti_(3)C_(2)T_(x)MXene nanoparticles(NPs)(PCM matrices),and explored their myogenic potential for skeletal muscle tissue regeneration.The PCM matrices demonstrated favorable physicochemical properties,including structural uniformity,alignment,microporosity,and hydrophilicity.In vitro assays revealed that the PCM matrices promoted cellular behaviors and myogenic differentiation of C2C12 myoblasts.Moreover,in vivo experiments demonstrated enhanced muscle remodeling and recovery in mice treated with PCM matrices following VML injury.Mechanistic insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices,leading to elevated intracellular Ca^(2+)levels in myoblasts through the activation of inducible nitric oxide synthase(i NOS)and serum/glucocorticoid regulated kinase 1(SGK1),ultimately promoting myogenic differentiation via the m TOR-AKT pathway.Additionally,upregulated i NOS and increased NO–contributed to myoblast proliferation and fiber fusion,thereby facilitating overall myoblast maturation.These findings underscore the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery.

Reproductive outcomes of water pipe smoking:A scoping review

Objective:To present an overview of the adverse effects of water pipe smoking on various aspects of reproduction and perinatal complications,covering all the most relevant studies,from descriptive findings to clinical trials.Methods:We reviewed the literature and included full publications in English provided by international biomedical databases,including PubMed,Science Direct,EBSCO,Scopus,and Google Scholar,and Cochrane Library over the last 38 years(May 1985 to June 2023).Search terms(Water pipe,Narghile,Shisha,Dokha,etc.,and Reproduction,Fertility,etc.)were adopted from Medical Subheading(MeSH)and Boolean operators were utilized to improve sensitivity.Results:133 Papers were identified in the initial search,of which 29 were excluded due to duplication.Eliminating search overlap and irrelevant records by reviewing titles and abstracts reduced 42 papers.62 Full-publication articles met the inclusion criteria.We summarized the effects of waterpipe smoke exposure on reproductive health based on three main categories:male and female fertility,fetal and neonatal outcomes,and pregnancy complications.Conclusions:Water pipe smoking has potential adverse effects on different aspects of human reproductive health,including semen parameters,male and female fertility,and fetal and neonatal health,and contributes to many maternal complications and morbidities.

Changes in Health Consciousness of Nursing Students in Japan after Acquiring Medical Care Knowledge from a Nursing School

Background: Health consciousness in daily life is very important because lifestyle habits, such as dietary and exercise habits, sleep, and smoking, are involved in the development and progression of lifestyle-related diseases, such as cancer, heart diseases, stroke, liver diseases, kidney diseases, and diabetes. Purpose: In this study, we surveyed to see if the health consciousness increased and what behaviors in daily life changed in students of a nursing school (Seiwa Nursing College, Tokyo, Japan) after they learned medical care knowledge required to become a nurse. Results: After learning medical care knowledge for 3 years, the students were found to have a higher level of health consciousness in the third year than they did in the first year. They applied the medical care knowledge they learned in daily life. All data were collected in 2019. Conclusions: The medical care knowledge learned in a nursing school was demonstrated to serve as information to increase the health consciousness.

Multidisciplinary therapy for treatment of patients with peritoneal carcinomatosis from gastric cancer

There is no standard treatment for peritoneal carcinomatosis (PC) from gastric cancer.A novel multidisciplinary treatment combining bidirectional chemotherapy [neoadjuvant intraperitoneal-systemic chemotherapy protocol (NIPS)],peritonectomy,hyperthermic intraperitoneal chemoperfusion (HIPEC) and early postoperative intraperitoneal chemotherapy has been developed.In this article,we assess the indications,safety and eff icacy of this treatment,review the relevant studies and introduce our experiences.The aims of NIPS are stage reduction,the eradication of peritoneal free cancer cells,and an increased incidence of complete cytoreduction (CC-0) for PC.A complete response after NIPS was ob-tained in 15 (50%) out of 30 patients with PC.Thus,a signif icantly high incidence of CC-0 can be obtained in patients with a peritoneal cancer index (PCI) ≤ 6.Using a multivariate analysis to examine the survival benef it,CC-0 and NIPS are identified as significant indicators of a good outcome.However,the high morbidity and mortality rates associated with peritonectomy and perioperative chemotherapy make stringent patient selection important.The best indications for multidisciplinary therapy are localized PC (PCI≤6) from resectable gastric cancer that can be completely removed during a peritonectomy.NIPS and complete cytoreduction are essential treatment modalities for improving the survival of patients with PC from gastric cancer.

Single injection of platelet-rich plasma as a novel treatment of carpal tunnel syndrome

Both in vitro and in vivo experiments have confirmed that platelet-rich plasma has therapeutic effects on many neuropathies, but its effects on carpal tunnel syndrome remain poorly understood. We aimed to investigate whether single injection of platelet-rich plasma can improve the clinical symptoms of carpal tunnel syndrome. Fourteen patients presenting with median nerve injury who had suffered from mild carpal tunnel syndrome for over 3 months were included in this study. Under ultrasound guidance, 1-2 m L of platelet-rich plasma was injected into the region around the median nerve at the proximal edge of the carpal tunnel. At 1 month after single injection of platelet-rich plasma, Visual Analogue Scale results showed that pain almost disappeared in eight patients and it was obviously alleviated in three patients. Simultaneously, the disabilities of the arm, shoulder and hand questionnaire showed that upper limb function was obviously improved. In addition, no ultrasonographic manifestation of the carpal tunnel syndrome was found in five patients during ultrasonographic measurement of the width of the median nerve. During 3-month follow-up, the pain was not greatly alleviated in three patients. These findings show very encouraging mid-term outcomes regarding use of platelet-rich plasma for the treatment of carpal tunnel syndrome.

Role of surgery in colorectal liver metastases:Too early or too late?

As colorectal cancer and colorectal liver metastases become a serious public health problem,new treatment modalities are needed in order to achieve better results. In the last decade there has been very important progress in oncology,with new and more effective chemotherapeutic agents administered alone or in combination improving the resectability rate in up to 40%of patients with colorectal liver metastases.Advances in interventional radiology,in particular,with the use of portal vein embolization and radiofrequency thermal ablation are new strategies allowing major liver resections and treatment of small liver metastases or early recurrences.Surgery,however,remains the gold standard strategy with intention to treat.In this review article we will describe the advanced role of surgery in the multidisciplinary approach to colorectal liver metastases,and the clinical problems the liver surgeon has to deal with,such as theresectability of the metastases,the presence of bilobar liver lesions and extrahepatic disease,the impact of chemotherapy in already resectable liver metastases,the problem of vanishing metastases after chemotherapy and the dilemma of staged or combined liver and colon operations and which organ first in the clinical scenario of synchronous colorectal liver metastases.

Early Aerobic Exercise Promotes Neurological Function Recovery of Rats after Cerebral Ischemia/Reperfusion by Upregulating the Expression of Heat Shock Protein A5

Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from cerebral ischemia/reperfusion and to determine whether these effects are associated with the expression level of HSPA5 in the ischemic penumbra.Methods A total of 72 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group[middle cerebral artery occlusion(MCAO)-Ex,n=18],ischemia and sedentary group(MCAO-St,n=18),sham-surgery and exercise group(Sham-Ex,n=18),or sham-surgery and sedentary group(Sham-St,n=18).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 min,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 min once a day for 5 consecutive days.After reperfusion,the motor function of the rats was scored by the Bederson neurological function test,balance beam test,and screen test.Nissl staining was conducted to assess morphological and structural change of nerve cells in the ischemic penumbra.The reverse transcription-quantitative polymerase chain reaction was applied to detect the mRNA expression of HSPA5.Western blot analysis was conducted to determine the protein expression of HSPA5.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was carried out in the ischemic penumbra after MCAO.Results Rats receiving early treadmill exercise had lower Bederson neurological function,balance beam,and screen test scores on the 3rd,7th,and 14th days after MCAO;in addition,more neurons survived in the ischemic penumbra after MCAO,and higher mRNA and protein expression of HSPA5 and fewer TUNEL-positive stained cells were observed.Conclusion Our study demonstrated that early aerobic exercise can improve neurological function recovery after ischemia/reperfusion.Furthermore,the increased level of HSPA5 in the ischemic penumbra might be one of the mechanisms of enhanced neurological function recovery.

Expression of endothelial nitric oxide synthase and vascular endothelial growth factor in association with neovascularization in human primary astrocytoma

Objective: To investigate the relationship between the expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF) and angiogenesis in primary astrocytoma. Methods: Thirty-seven primary astrocytomas and 4 astrocytic hyperplasia samples were collected and divided into three groups according to histological grade. The expression of eNOS, VEGF and factor Ⅷ related antigen (FVIIIRAg) were assayed by immunohistochemistry. Microvascular density was assessed by FVIIIRAg immunoreactivity. The intensity of immunoreactivity was graded according to the percentage of positive tumor cells. Results: No eNOS and VEGF were expressed in the astrocytes and vascular endothelium in astrocytic hyperplasia.The expression of eNOS or VEGF was light in low-grade astrocytoma and strong in glioblastoma. eNOS expression in astrocytoma was very positively correlated with VEGF. eNOS and VEGF expression in anaplastic astrocytoma was median in contrast to the low grade astrocytoma and glioblastoma. Lower microvascular density was found in low grade astrocytoma than that in higher grade malignant ones. The expressions of eNOS and VEGF were correlated with microvascular density and tumor malignancy.Conclusion: This finding suggests that eNOS and VEGF may have cooperative effect in tumor angiogenesis and play an important role in the pathogenesis of primary astrocytoma.

Neuroprotective effects of a mitochondrial K^+-ATP channel opener (diazoxide) are mediated by Bcl-2 expression upregulation

Mitochondrial K＋-ATP （mito-KATP） channels play an important role in cellular function and survival following ischemic stress. The present results revealed that intervention with diazoxide, a mito-KATP channel opener, led to an increase in Bcl-2 expression in the cerebral cortex of rats subjected to cerebral ischemia reperfusion injury. In addition, the intervention also led to clear improvements in neuronal mitochondrial morphology and consciousness post-injury. Glibenclamide a mito-KATP channel blocker, exhibited the converse effects. Both diazoxide and glibenclamide exerted dose-dependent effects （in particular, at 18 mg/kg diazoxide and 25 mg/kg glibenclamide）. These findings suggest that diazoxide exerts a neuroprotective effect on cerebral ischemia reperfusion injury by opening mito-KATP channels and upregulating Bcl-2 expression.

Some of the experimental and clinical aspects of the effects of the maternal diabetes on developing hippocampus

Diabetes mellitus during pregnancy is associated with an increased risk of multiple congenital anomalies in progeny.There are sufficient evidence suggesting that the children of diabetic women exhibit intellectual and behavioral abnormalities accompanied by modification of hippocampus structure and function.Although,the exact mechanism by which maternal diabetes affects the developing hippocampus remains to be defined.Multiple biological alterations,including hyperglycemia,hyperinsulinemia,oxidative stress,hypoxia,and iron deficiency occur in pregnancies with diabetes and affect the development of central nervous system(CNS) of the fetus.The conclusion from several studies is that disturbance in glucose and insulin homeostasis in mothers and infants are major teratogenic factor in the development of CNS.Insulin and Insulin-like growth factor-1(IGF-1) are two key regulators of CNS function and development.Insulin and IGF-1 receptors(IR and IGF1 R,respectively) are distributed in a highly specific pattern with the high density in some brain regions such as hippocampus.Recent researches have clearly established that maternal diabetes disrupts the regulation of both IR and IGF1 R in the hippocampus of rat newborn.Dissecting out the mechanisms responsible for maternal diabetes-related changes in the development of hippocampus is helping to prevent from impaired cognitive and memory functions in offspring.

Proteomic analysis of the dorsal spinal cord in the mouse model of spared nerve injury-induced neuropathic pain

Peripheral nerve injury often causes neuropathic pain and is associated with changes in the expression of numerous proteins in the dorsal horn of the spinal cord. To date, proteomic analysis method has been used to simultaneously analyze hundreds or thousands of proteins differentially expressed in the dorsal horn of the spinal cord in rats or dorsal root ganglion of rats with certain type of peripheral nerve injury. However, a proteomic study using a mouse model of neuropathic pain could be attempted because of abundant protein database and the availability of transgenic mice. In this study, whole proteins were extracted from the ipsilateral dorsal half of the 4th-6th lumbar spinal cord in a mouse model of spared nerve injury（SNI）-induced neuropathic pain. In-gel digests of the proteins size-separated on a polyacrylamide gel were subjected to reverse-phase liquid-chromatography coupled with electrospray ionization ion trap tandem mass spectrometry（MS/MS）. After identifying proteins, the data were analyzed with subtractive proteomics using ProtAn, an in-house analytic program. Consequently, 15 downregulated and 35 upregulated proteins were identified in SNI mice. The identified proteins may contribute to the maintenance of neuropathic pain,and may provide new or valuable information in the discovery of new therapeutic targets for neuropathic pain.

Protective Effect of Emodin against Lipopolysaccharides-induced Corneal Injury in Rats

Objective To investigate the effect of emodin on lipopolysaccharides(LPS)-induced corneal injury in rats.Methods Three parallel incisions on the central surface of corneal epithelium were made and LPS was applied on them to induce corneal injury in Wistar rats.All rats were randomly divided into emodin group(n=40) and keratitis group(n=40).Rats in the emodin group received subconjunctival injection of emodin and rats in the keratitis group received its vehicle 30 minutes before LPS exposure.At different time points-1,3,6,12,and 24 hours after LPS exposure,the symptoms of all rats were observed and the severity of their ocular inflammation was examined with a slit lamp microscope,then 8 rats in each group were killed through cervical dislocation and their eyes were enucleated and prepared to observe pathological changes of corneal tissue under a light microscope.The activation of nuclear factor-κB(NF-κB) under different conditions was determined by Western blot.Immunocytochemistry staining with an antibody against intercellular adhesion molecule-1(ICAM-1) was performed to identify positive cells in corneal tissues.Results The model of acute keratitis was successfully established in Wistar rats.LPS could induce a typical corneal inflammatory response,such as hyperemia,corneal edema and opacity,which were observed in model rats.Compared with keratitis group,both ocular behaviors and damages of the corneal structure were improved in emodin group.Furthermore,the activation of NF-кB and the expression of ICAM-1 induced by LPS were markedly inhibited in emodin group.Conclusion Emodin can inhibit the activation of NF-κB and the expression of ICAM-1 induced by LPS in corneas,protect against acute corneal injury,and improve symptoms in rats.

Dual effects of 3,4-methylenedioxymethamphetamine (ecstasy) on survival and apoptosis of primary hippocampal neurons

BACKGROUND： 3, 4-methylenedioxymethamphetamine （MDMA, also known as ＂ecstasy＂） has been shown to exhibit neurotoxic effects on the hippocampus. However, exposure to sub-lethal insults of MDMA has been reported to result in neuroprotection. OBJECTIVE： To investigate the effects of MDMA on hippocampal neuronal viability, caspase-3 activity, and mRNA expression of the N-methyI-D-aspartate （NMDA） receptor 2B （NR2B） subunit. DESIGN, TIME AND SETTING： A cytological, in vitro experiment was performed at the Department of Anatomy, School of Medicine, and Department of Toxicology-Pharmacology, Faculty of Pharmacy Tehran University of Medical Sciences in 2008. MATERIALS： MDMA was extracted from ecstasy tablets, which were kindly supplied by the Pharmacology-Toxicology Department, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran. METHODS： Hippocampal neurons were isolated from Wistar rats at gestational day 18. Following primary culture, hippocampal neuronal viability was detected by MTT assay. Varying concentrations of MDMA （100-5 000 μmol/L） were used to determine lethal concentration 50 （LC50）, which was around 1 500 μmol/L. Five concentrations of MDMA below 1 500 μmol/L （100, 200, 400, 800, and 1 050 μmol/L） were used for the remaining experiments. After 24 hours of MDMA treatment, NR2B mRNA expression was detected by RT-PCR, and caspase-3 relative activity was determined by colorimetric assay. MAIN OUTCOME MEASURES： Hippocampal neuronal viability, caspase-3 activity, and NR2B mRNA expression. RESULTS： MDMA-induced neurotoxicity in hippocampal neuronal cultures was dose-dependent. In high concentrations （1 000-5 000μmol/L） of MDMA, neuronal viability was decreased. However, with a 500 μmol/L dose of MDMA, neuronal viability was significantly increased （P 〈 0.01）. Low concentrations of MDMA （200 and 400μmol/L） significantly decreased caspase-3 activity （P 〈 0.01）, whereas high concentrations of MDMA significantly increased caspase-3 activity （P 〈 0.01）. NR2B subunit mRNA expression was not significantly altered after 100 -1 050 μmol/L MDMA exposure. CONCLUSION： MDMA exhibits dual effects on hippocampal neuronal viability and caspase-3 activity. These effects are independent from NR2B subunit expression levels.

Neurochemical mechanism of the gastrointestinal interdigestive migrating motor complex in rats with acute inflammatory stomach ache

The normal gastrointestinal interdigestive migrating motor complex cycle was interrupted, and paroxysmal contraction appeared after formaldehyde-induced stomach ache. Activities of nitric oxide synthase, acetylcholinesterase and vasoactive intestinal peptide neurons were significantly reduced, whereas activities of calcitonin gene-related peptide neurons were significantly increased in the pyloric sphincter muscular layer, myenteric nerve plexus and submucous nerve plexus. Electroacupuncture at Zusanfi （ST36） suppressed paroxysmal contraction in rats with formaldehyde-induced stomach ache, and neurons in the enteric nervous system were normal. These results indicated that nitrergic neurons, cholinergic neurons, vasoactive intestinal peptide neurons and calcitonin gene-related peptide neurons in the enteric nervous system may be involved in changes to the gastrointestinal interdigestive migrating motor complex following stomach ache, and that electroacupuncture can regulate this process.

Radiological evaluation of patellofemoral instability and possible causes of assessment errors

Patellofemoral instability(PI)is the disruption of the patella’s relationship with the trochlear groove as a result of abnormal movement of the patella.To identify the presence of PI,conventional radiographs(anteroposterior,lateral,and axial or skyline views),magnetic resonance imaging,and computed tomography are used.In this study,we examined four main instability factors:Trochlear dysplasia,patella alta,tibial tuberosity–trochlear groove distance,and patellar tilt.We also briefly review some of the other assessment methods used in the quantitative and qualitative assessment of the patellofemoral joint,such as patellar size and shape,lateral trochlear inclination,trochlear depth,trochlear angle,and sulcus angle,in cases of PI.In addition,we reviewed the evaluation of coronal alignment,femoral anteversion,and tibial torsion.Possible causes of error that can be made when evaluating these factors are examined.PI is a multi-factorial problem.Many problems affecting bone structure and muscles morphologically and functionally can cause this condition.It is necessary to understand normal anatomy and biomechanics to make more accurate radiological measurements and to identify causes.Knowing the possible causes of measurement errors that may occur during radiological measurements and avoiding these pitfalls can provide a more reliable road map for treatment.This determines whether the disease will be treated medically and with rehabilitation or surgery without causing further complications.

Variations in the Course of the Inferior Gluteal Nerve and Artery: A Case Report and Literature Review

Variations in the course of the inferior gluteal nerve and artery were observed in Japanese cases (4/94 sides). In these variation cases, the inferior gluteal nerve exited the pelvis from the upper edge of the piriformis (suprapiriformis foramen) in 4/4 sides (4.26%). In 2/4 sides (2.13%), the normal inferior gluteal artery was not observed, except that a fine artery exited the pelvis from the inferior piriformis foramen to form an “arch” with the superior gluteal artery under the gluteal maximus in 1/4 side. Moreover, in 1/4 side, a twig of the internal pudendal artery exited pelvis from inferior piriformis foramen and distributed to the surrounding tissues. The present observations of the inferior gluteal nerve and artery course are very important and useful for surgeons and nurses.

Comparison of Chondrogenic Ability between Mesenchymal Stem Cells and Buffy Coat <i>in Vitro</i>

Objective: In vitro comparison of chondrogenic differentiation ability of bone marrow extracted mesenchymal stem cells (MSCs) and buffy coat. Methods: MSCs of New Zealand white rabbits were cultured in vitro and adherent cells were passaged. The cells were inoculated on polyglycolic acid (PGA) scaffold (3 mm in diameter and 2 mm in height) liquid for 21 days. Under the same conditions, without subculture, buffy coat was directly inoculated into the cell scaffold. The degree of chondrogenic differentiation was compared by Safranin-Ostaining, histological scoring and biochemical functional detection. Results: The chondrogenic differentiation ability of the buffy coat group was better than that of the MSC group. Safranin-Ostain was stronger in buffy coat group than in MSC group. The Bern Score was also higher in the buffy coat group than in the MSC group and the total amount of sulfated glycosaminoglycans (GAGs) in buffy coat group was higher than that in MSC group. Conclusion: The chondrogenic differentiation ability of buffy coat is higher than that of MSC. Through this result, it was found buffy coat can also differentiate into sub-cartilage without stimulating growth factor. Thus, buffy coat can make a great source in artificial cartilage engineering.

Anti-Ulcerogenic Activity of the Pomegranate Peel (<i>Punica granatum</i>) Methanol Extract

Pomegranate (Punica granatum L.) belongs to genera Punica and family Punicaceae. It is a herbal preparation that has been suggested as useful in the treatment of gastrointestinal disorders. However, to our knowledge, no study has been conducted to evaluate this therapeutic property. In the present study the antiulcerogenic effects of pomegranate peel methanol extract, was tested on male Wistar albino rats. Oral pretreatment with peel extracts (25, 50 and 100 mg/kg) for 15 days protected the gastric mucosa against the damage induced by indomethacin (50 mg/kg). The incidence of ulceration in the control group was 100%. The best results were found in a dosage of 50 mg/kg in sour summer cultivar which inhibited the peptic ulcerin comparison with indomethacin induced gastric ulcer group. Lowest ulcer index (5.4 ± 0.55), an apparent decrease in the infiltration of polymorphonuclear leukocytes and hemorrhage were observed after administration of sour summer extracts (50 mg/kg). In conclusion present study showed that pomegranate peel extract, especially sour summer, has curative potential as an antiulcer, possibly via its high antioxidant activity. These results from pomegranate peel extract can provide an extra income and may contribute about good nutritional values of this product.

Contribution of mammalian target of rapamycin in the pathophysiology of cirrhotic cardiomyopathy

AIM: To explore the role of mammalian target of rapamycin(m TOR) in the pathogenesis of cirrhotic cardiomyopathy and the potential of rapamycin to improve this pathologic condition.METHODS: Male albino Wistar rats weighing 100-120 g were treated with tetrachloride carbon(CCl_4) for 8 wk to induce cirrhosis. Subsequently, animals were administered rapamycin(2 mg/kg per day). The QT_c intervals were calculated in a 5-min electrocardiogram. Then, the left ventricular papillary muscles wereisolated to examine inotropic responsiveness to β-adrenergic stimulation using a standard organ bath equipped by Powerlab system. Phosphorylated-m TOR localization in left ventricles was immunohistochemically assessed, and ventricular tumor necrosis factor(TNF)-α was measured. Western blot was used to measure levels of ventricular phosphorylated-m TOR protein.RESULTS: Cirrhosis was confirmed by hematoxylin and eosin staining of liver tissues, visual observation of lethargy, weight loss, jaundice, brown urine, ascites, liver stiffness, and a significant increase of spleen weight(P < 0.001). A significant prolongation in QTc intervals occurred in cirrhotic rats exposed to CCl_4(P < 0.001), while this prolongation was decreased with rapamycin treatment(P < 0.01). CCl_4-induced cirrhosis caused a significant decrease of contractile responsiveness to isoproterenol stimulation and a significant increase in cardiac TNF-α. These findings were correlated with data from western blot and immunohistochemical studies on phosphorylated-m TOR expression in left ventricles. Phosphorylated-m TOR was significantly enhanced in cirrhotic rats, especially in the endothelium, compared to controls. Rapamycin treatment significantly increased contractile force and myocardial localization of phosphorylated-m TOR and decreased cardiac TNF-α concentration compared to cirrhotic rats with no treatment. CONCLUSION: In this study, we demonstrated a potential role for cardiac m TOR in the pathophysiology of cirrhotic cardiomyopathy. Rapamycin normalized the inotropic effect and altered phosphorylated-m TOR expression and myocardial localization in cirrhotic rats.

Antioxidant effects of quercetin in freeze-thawing process of mouse spermatogonialstem cells

Objective: To evaluate the antioxidant effect of quercetin on cell viability, reactive oxygen species (ROS) contents and apoptosis of cryopreserved mouse spermatogonial stem cells (mSSCs). Methods: mSSCs were isolated from neonate mice and cultivated in culture medium containing 30 μM quercetin for 48 h and then frozen for 2 weeks. After thawing, MTT assay was carried out to analyze the cell viability. Moreover, intracellular ROS levels were measured by flow cytometery and apoptosis was evaluated by detection of phosphatidylserine externalization assay and also real-time polymerase chain reaction. Results: Pre-treatment of mSSCs by 30 μM quercetin significantly decreased intracellular ROS content and apoptotic cell numbers and improved viability of mSSCs. Moreover, the gene expression of Bcl-2 and Bax significantly increased and decreased respectively after the freeze-thawing process. Conclusions: Pre-treatment of mSSCs with quercetin can improve cell viability and reduce apoptosis during freeze-thawing process. It can be a promising way to improve the quality and efficiency of cryopreservation protocols used in fertility preservation strategies.