Puerarin protects the fatty liver from ischemia-reperfusion injury by regulating the PI3K/AKT signaling pathway

Abstract The incidence of non-alcoholic fatty liver (NAFLD) remains high, and many NAFLD patients suffer from severe ischemia-reperfusion injury (IRI). Currently, no practical approach can be used to treat IRI. Puerarin plays a vital role in treating multiple diseases, such as NAFLD, stroke, diabetes, and high blood pressure. However, its role in the IRI of the fatty liver is still unclear. We aimed to explore whether puerarin could protect the fatty liver from IRI. C57BL/6J mice were fed with a high‐fat diet (HFD) followed by ischemia reperfusion injury. We showed that hepatic IRI was more severe in the fatty liver compared with the normal liver, and puerarin could significantly protect the fatty liver against IRI and alleviate oxidative stress. The PI3K-AKT signaling pathway was activated during IRI, while liver steatosis decreased the level of activation. Puerarin significantly protected the fatty liver from IRI by reactivating the PI3K-AKT signaling pathway. However, LY294002, a PI3K-AKT inhibitor, attenuated the protective effect of puerarin. In conclusion, puerarin could significantly protect the fatty liver against IRI by activating the PI3K-AKT signaling pathway.

Case reports of two children with neurofibromatosis type 1 / 中国热带医学

@#Abstract: Objective　To report the clinical characteristics and genetic test results of two children with neurofibromatosis type 1 (NF1), and to provide reference for the comprehensive diagnosis-treatment and follow-up plans of NF1 patients based on the existing diagnosis and treatment progress of NF1. Methods　Two children with NF1 admitted to the Department of Children's Medicine, Haikou people's Hospital in May and June 2022 were selected to analyze the clinical data of their clinical manifestations, laboratory examination, genetic test results, diagnosis and treatment and follow-up retrospectively. Results　Two children had typical clinical manifestations, such as café-au-lait spots, axillary freckles, intraocular iris hamartoma. Venous blood was collected from case 1 and his parents for NF gene test, and a new mutation of c.4084C>T in the NF1 gene was found, and their parents did not have the pathogenic gene; the venous blood of the children in case 2 was tested for whole-exome gene analysis, and a heterozygous nonsense variant c.910C>T:p.R304 on the NF1 gene was found, , which was verified by Sanger sequencing to be inherited from his mother, his mother has café-au-lait spots and brain glioma, and has undergone surgery to remove the brain glioma, but has not undergone chemoradiotherapy or targeted therapy. No neurological malignancies were detected in either of the two children at follow-up until July 2022. Conclusions　The clinical manifestations of NF1 are relatively typical, genetic testing is conducive to determine its classification, and regular follow-up review can help to detect and treat malignant tumors early, thus improving the patient's quality of life.

Encephalopathy due to defective mitochondrial and peroxisomal fission-1 a case report / 中国热带医学

@#Abstract： Objective　To investigate the clinical characteristics and pathogenic genetic mutation of a case with encephalopathy due to defective mitochondrial and peroxisomal fission-1 (EMPF1). Methods　The clinical data and genetic test results of a patient with EMPF1 admitted to the Department of Pediatrics, the Affiliated Hospital of Xiangya Medical College of Central South University in August 2020 were retrospectively analyzed. Results　An 8-year-old girl, her main clinical features were developmental regression, microcephaly, hypotonia, refractory epilepsy, cranial MRI suggesting brain atrophy and abnormal signals in the right temporal-occipital-parietal cortex, aEEG showing slow wave discharge in the right hemisphere; Whole-exome sequencing of families suggested that the child had a heterozygous missense variant at the c.1040C>G site in the DNM1L gene and the verification results by Sanger sequencing showed that her parents had no variant in this site, which was a novel mutation in accordance with autosomal dominant inheritance; bioinformatics analysis predicted that the mutation was pathogenic. After 2 years of outpatient follow-up, the patient's condition was stable after mitochondrial cocktail therapy and antiepileptic drugs, no epileptic seizure occurred in the past year, mental state and swallowing function improved, and she could be fed orally with occasional nausea and vomiting. Conclusions　The main clinical manifestations of EMPF1 are psychomotor developmental delay or regression, dystonia, limb paralysis, epilepsy and so on. According to the clinical phenotype and genetic test results, the rare disease can be diagnosed early.

Clinical analysis of elven families with mitochondrial encephalomyopathy in children / 中国热带医学

@#Abstract: Objective To investigate the clinical phenotype and genotype characteristics of mitochondrial encephalomyopathy (ME) families in children. Methods　The clinical data and genetic test results of eleven ME families who were admitted to the department of pediatrics of three tertiary hospitals in Hainan Province from January 2007 to December 2021 were retrospectively analyzed. Results　A total of 13 cases were diagnosed in eleven ME families, including 6 males (46.15%) and 7 females (53.85%). The age of onset ranged from 6 months to 12 years, the interval from onset to diagnosis was 9 months to 8 years and Morava score was 6-11. Clinical symptoms mainly included abnormal movement, developmental retardation or regression, seizures, stroke-like episodes; among the 13 children, 11 (84.62%) had elevated blood lactic acid and 4 (30.77%) had elevated blood creatine kinase. Cranial MRI mainly involved temporal parietal occipital lobe, cerebellum, brainstem and basal ganglia, some with brain atrophy. Gene detection showed that 8 families (72.72%) were caused by mtDNA mutation, of which 5 families and 6 patients were caused by MT-TL1, m.3243A>G, and 5 asymptomatic carriers of 4 families (80.00%) were detected; MT-ND5, m.13513 G>A was detected in 2 families and 3 patients, and an asymptomatic mutation carrier was detected in a family (50.00%); MT-ND3, m.10191T>C was detected in one family and one patient, and 2 asymptomatic mutation carriers were detected. Three families were caused by nDNA mutations (27.27%). A compound heterozygous mutation of c.751C>T and c.516-2A >G in SURF1 gene was found in one family and one patient, which followed autosomal recessive inheritance. The pathogenic loci were inherited from mother and father, respectively. Two new spontaneous mutations c.1040C>G and c.2060_2062delTAG in DNM1L gene were respectively detected in two families and two patients. All children were given mitochondrial cocktail therapy and symptomatic treatment after diagnosis by genetic testing. Follow-up to June 2022, two families were lost to follow-up and 9 families were followed up regularly; three of the 11 children were still survived. Conclusions　For children diagnosed with ME, genetic testing of family members can screen out early asymptomatic pathogenic mutation carriers, achieve early diagnosis of ME and guide clinical genetic counseling. Two new pathogenic sites of DNM1L gene were found in this study, which expanded the genotype spectrum.

Genetic analysis of a female child family with suspected X-linked recessive genetic disease / 中国热带医学

@#Objective　Genetic analysis was performed on a female child with chromosome Xq28 heterozygous deletion and suspected X-linked recessive disease to determine the morbidity and prognosis. Methods　A female child was admitted to the hospital on day 20 because of "jaundice for 20 days and difficulty in stopping bleeding at acupuncture sites". Low depth whole genome test of amniocentesis in late pregnancy suggested missing copy number of hemophilia A and X-linked mental retardation type 72. In order to further confirm the diagnosis and prognosis, peripheral blood of the children and their parents were collected for gene testing, chromosome inactivation test and genetic analysis. Results　Chromosome Xq28 of the child had 439.4 kb copy number heterozygous deletion variation, which was a clear disease-coding gene for functional loss included in ClinGen database. Chromosome inactivation test showed that the paternal X chromosome of the child was extremely inactivated. Haplotype analysis suggested that the normal chromosome of the subject was inherited from the mother, and there was heterozygous deletion on the paternal X chromosome, so it was inferred that the child will not develop disease or just have mild symptoms. Conclusion　It is necessary to analyze the X chromosome inactivation test for female patients with the pathogenic variation of X-linked recessive genetic disease to determine the possibility of the disease.

Mitochondrial combined oxidative phosphorylation deficiency type 1 in childrena case report and literature review / 中国热带医学

@#Objective　To summarize the phenotypic and genotypic characteristics of mitochondrial combined oxidative phosphorylation deficiency type 1 (COXPD1), and to improve the clinicians' awareness of this mitochondrial encephalomyopathy. Methods　The clinical characteristics, physical examination, laboratory examination and other data of a child with COXPD1 were analyzed retrospectively. The diagnosis was confirmed by clinical whole exon sequencing and high-precision mitochondrial genome full-length PLUS gene detection, and the phenotype and genotype were analyzed by reviewing relevant literature. Results　A one-year and five-month-old boy mainly presented with hyperlactacidemia and abnormal liver function. Clinical whole exon sequencing showed that the child had homozygous variation of c. 688G>A(p.G230S) in the GFM1 gene. Sanger sequencing verified that the variation was respectively inherited from the parents of the child (both were heterozygous) with the autosomal recessive inheritance pattern. The high-precision mitochondrial genome full-length PLUS detection also did not find pathogenic mutations related to clinical phenotypes. The child was diagnosed with COXPD1. After "cocktail" therapy and liver protection therapy, the patient's condition improved. Conclusions　The phenotype of COXPD1 is complicated and variable, mainly liver type and brain type. The mutation of GFM1 gene affects mitochondrial translation system function, and early gene detection is helpful for definite diagnosis.

Comments on research status and future of rare diseases in China / 中国热带医学

@#Rare diseases, also known as "orphan diseases", refer to diseases with very low incidence. Countries and regions define rare diseases according to epidemiological standards, economic standards of rare drugs and disease severity. The World Health Organization (WHO) has suggested the prevalence rate of less than 6.5 to 10 per 10 000 people to define rare diseases. In May 2018, "China's First List of Rare Diseases" was released, including 121 rare diseases. Most rare diseases are hereditary diseases with early onset, severe disease, and poor prognosis. About 75% of rare genetic diseases occur in the neonatal period or childhood, which are important part of human birth defects and brings a huge burden to society and families. The effective prevention and treatment of rare diseases is one of the important goals of building a "Healthy China". With the development of molecular biology technology and the continuous research and development of advanced medical products in the field of gene therapy, the level of clinical diagnosis and treatment of rare diseases has risen to a new level, which provides a possibility for the cure of some rare diseases. In China, most rare diseases rely on imported drugs, which cost a lot and bring heavy economic burden to patients. Improving the medical insurance system for rare diseases has become a difficult point in the current medical reform. This paper mainly discusses the definition of rare diseases, the research status, efforts and future development direction of rare diseases in China, in order to deepen the understanding and response of medical workers and the whole society to rare diseases.

Analysis of personalized precision diagnosis of 156 children with suspected rare inherited metabolic diseases / 中国热带医学

@#Abstract: Objective　To explore the accurate diagnosis of children with suspected rare inherited metabolic diseases, and to compare the application value of mass spectrometry and genetic testing in the diagnosis of rare inherited metabolic diseases (IMD). Methods　The clinical information, mass spectrometry, and genetic results of children with suspected rare inherited metabolic diseases admitted to the Department of Pediatrics, the Affiliated Haikou Hospital of Xiangya Medical College, Central South University from March 2017 to December 2021 were analyzed retrospectively. Results　156 children with suspected rare inherited metabolic diseases were detected by mass spectrometry, 67 cases were positive and 89 cases were negative. Children with positive initial examination were retested, and 19 cases were positive. Among the retest positive cases, 13 cases were given genetic testing, and 9 cases were positive and 4 cases were negative. Among the initial negative cases, 54 children with poor therapeutic effect and high clinical suspicion of inherited metabolic diseases completed genetic testing, 15 cases were positive and 39 cases were negative. The results of the two detection methods were compared, the positive rate of mass spectrometry was 19.4%(13/67), and the positive rate of genetic testing was 35.8%(24/67). The continuity correction of Pearson's chi-square test of continuity correction suggested that the results of genetic testing and mass spectrometry were different, and the difference was statistically significant (P<0.05). Taking genetic testing as the gold standard, the sensitivity and specificity of mass spectrometry detection were 37.5% (95%CI:19.6%-59.2%) and 90.7% (95%CI:76.9%-97.0%), respectively. Among the 24 confirmed cases, 5 cases were diagnosed by gene panel and 19 cases were diagnosed by whole exome sequencing (WES). One case diagnosed by WES had no pathogenic mutation detected by gene panel before diagnosis. The detection of DNM1L gene c.1040C>G and AMN gene c.651+1G>C are novel pathogenic gene variants, which have clinical significance. Conclusions　The ability of mass spectrometry in the diagnosis of inherited metabolic diseases is limited. Genetic testing, especially whole exome sequencing, can be the first choice for individualized diagnosis of suspected rare inherited metabolic diseases. In addition, the new mutation sites found by WES in this study enriched the pathogenic gene mutation spectrum and provided direction for further functional biological experiments.

Study of nonlinear viscoelastic mechanical properties of degenerated intervertebral discs / Estudio de las propiedades mecánicas viscoelásticas no lineales de discos intervertebrales degenerados

SUMMARY: Biomechanical factors are important factors in inducing intervertebral disc degeneration, in this paper, the nonlinear viscoelastic mechanical properties of degenerated intervertebral discs were analyzed experimentally. Firstly, the loading and unloading curves of intervertebral discs before and after degeneration at different strain rates were compared to analyze the changes of their apparent viscoelastic mechanical properties; The internal stress/strain distribution of the disc before and after degeneration was then tested by combining digital image technology and fiber grating technology. The results show that the intervertebral disc is strain-rate- dependent whether before or after degeneration; The modulus of elasticity and peak stress of the degenerated disc are significantly reduced, with the modulus of elasticity dropping to 50 % of the normal value and the peak stress decreasing by about 55 %; Degeneration will not change the distribution of the overall internal displacement of the intervertebral disc, but has a greater impact on the superficial and middle AF; The stress in the center of the nucleus pulposus decreases, and the stress in the outer AF increases after degeneration. Degeneration has a great impact on the nonlinear viscoelastic mechanical properties of intervertebral disc, which has reference value for the mechanism, treatment and prevention of clinical degenerative diseases.

RESUMEN: Los factores biomecánicos son importantes en la inducción de la degeneración del disco intervertebral. En este estudio se analizaron experimentalmente las propiedades mecánicas viscoelásticas no lineales de los discos intervertebrales degenerados. En primer lugar se compararon las curvas de carga y descarga de los discos intervertebrales, antes y después de la degeneración, a diferentes velocidades de deformación para analizar los cambios aparentes de sus propiedades mecánicas viscoelásticas. La distribución interna de tensión/deformación del disco antes y después de la degeneración se probó luego combinando tecnología de imagen digital y tecnología de rejilla de fibra. Los resultados mostraron que el disco intervertebral depende de la velocidad de deformación antes o después de la degeneración; El módulo de elasticidad y la tensión máxima del disco degenerado se reducen significativamente, cayendo el módulo de elasticidad al 50 % del valor normal y la tensión máxima disminuyendo en aproximadamente un 55 %; La degeneración no cambiará la distribución del desplazamiento interno general del disco intervertebral, pero tiene un mayor impacto en la FA superficial y media; El estrés en el centro del núcleo pulposo disminuye y el estrés en el FA externo aumenta después de la degeneración. La degeneración tiene un gran impacto en las propiedades mecánicas viscoelásticas no lineales del disco intervertebral, que tiene valor de referencia para el mecanismo, tratamiento y prevención de enfermedades clínicas degenerativas.

Research progress in association of neurological disorders and periodontal diseases / 中华口腔医学杂志

Periodontal diseases are inflammatory diseases caused by oral pathogens around the periodontal supporting tissues, leading to systemic and chronic inflammatory conditions. The continuous chronic systemic inflammation may be a trigger of neuroinflammation, which is the prominent feature of a variety of neurological disorders. It implies that there may be a causal link between periodontal diseases and neurological disorders. This article presents epidemiological and biological evidences that periodontal diseases can induce or exacerbate neurological disorders, including Alzheimer's disease, Parkinson's disease, multiple sclerosis and major depressive disorder, and analyzes the possible mechanisms. The importance of maintaining oral health as well as preventing and treating periodontal diseases are emphasized. At the same time, this may provide novel approaches to study the relationship between periodontal diseases and neurological disorders in the prevention and treatment strategies of neurological disorders.

Silencing of circular RNA ANRIL attenuates oxygen-glucose deprivation and reoxygenation-induced injury in human brain microvascular endothelial cells by sponging miR-622

BACKGROUND: Circular RNA (circRNA) is highly expressed in the brain tissue, but its molecular mechanism in cerebral ischemia-reperfusion remains unclear. Here, we explored the role and underlying mechanisms of circRNA antisense non-coding RNA in the INK4 locus (circ_ANRIL) in oxygen-glucose deprivation and reoxygenation (OGD/R)-induced cell injury. RESULTS: The expression of circ_ANRIL in OGD/R-induced human brain microvascular endothelial cells (HBMECs) was significantly up-regulated, while that of miR-622 was significantly down-regulated. Overexpression of circ_ANRIL significantly inhibited the proliferation of OGD/R-induced HBMECs and aggravated OGD/R-induced cell apoptosis. Moreover, circ_ANRIL overexpression further increased the secretion of interleukin (IL)-1ß, IL-6, tumor necrosis factor-a, and monocyte chemoattractant protein-1 in OGD/R-treated HBMECs. The results of bioinformatics analysis and luciferase reporter assay indicated that circ_ANRIL served as an miR-622 sponge to negatively regulate the expression of miR-622 in OGD/R-treated HBMECs. Additionally, circ_ANRIL silencing exerted anti-apoptotic and anti-inflammatory effects by positively regulating the expression of miR-622. Furthermore, inhibition of OGD/R-induced activation of the nuclear factor (NF)-kB pathway by circ_ANRIL silencing was significantly reversed by treatment with miR-622 inhibitor. CONCLUSIONS: Knockdown of circ_ANRIL improved OGD/R-induced cell damage, apoptosis, and inflammatory responses by inhibiting the NF-κB pathway through sponging miR-622.

Improvement effect of extract of St.John's wort tabIets on fear and anxiety behavior of post-traumatic stress disorder model mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of extract of St.John's wort tablets (ESJWT) on post-traumatic stress disorder (PTSD) in a mouse model and explore possible avenues for the treat-ment of PTSD.METHODS Forty-eight C57BL/6 male mice were randomly divided into 4 groups:normal control group, model group, sertraline and ESJWT treatment groups, with 12 mice in each group. Except the normal control group, all the mice were treated with fifteen intermittent inescapable foot-shocks (intensity:0.8 mA;interval:10 s;duration:10 s)for 2 d.Sertraline 15 mg·kg-1and ESJWT 25 mg·kg-1 were ig given one hour before foot-shocks repectively,once a day,for 18 d.Then,contextual freezing and fecal pellet were tested on the 3rd,8thand 15thdays.In addition,open field test,elevated plus maze test and staircase test were performed on the 16th, 17thand18thdays, respectively. Here, the day mice were subjected to short foot-shocks was defined as the 1stday.The serum contents of norepinephrine (NE) and 5-hydroxytryptamine (5-HT) were evaluated using ELISA after behavior tests. RESULTS Compared with normal control group, the freezing time and the number of fecal pellet in model group were significantly increased(P<0.01).Meanwhile,the time and number of entries into the central open field and open arm were decreased(P<0.01).The above experiments indicated that the mouse model of PTSD was established successfully. Compared with model group, both sertraline and ESJWT extract showed anti-PTSD effect, with the decreased percentage of freezing time (P<0.05, P<0.01). ESJWT extract treatment also reduced the amount of fecal pellet(P<0.01).However,no significant changes of the contents of NE and 5-HT in any group were seen.CONCLUSION ESJWT has anti-PTSD effect in the mouse model,which might be used for the treatment of PTSD.

Effect of ginsenoside Rg3 on mouse neural stem cell differentiation in vitro / 中国组织工程研究

BACKGROUND: Application of neural stem cells (NSCs) is of great current interest in neuroscience, but NSCs origin is very limited. And they always differentiate into a large percentage of glial cells and small percentage of neurons in natural differentiation process, so researchers should take effective measures to promote NSCs differentiation into certain offsprings. Previous studies have shown that ginseng saponin ingredients, such as Rb1 and Rg1, have certain influence on NSCs differentiation, but it is unclear whether Rg3 plays a role on NSCs differentiation. OBJECTIVE:To preliminarily investigate the effect of ginsenoside Rg3 on mouse NSCs differentiation into neurons and astrocytes in vitro. METHODS: The fetal cortices of embryonic 14 days (E14) C57BL/6 mice were isolated for culturing primary NSCs. Then passaged NSCs were identified by their purity with NSCs specific antibodies, Nestin and Sox2, by immunofluorescence staining. NSCs were induced for 3 days in the differentiation medium containing ginsenoside Rg3 of different concentrations (blank control, 50 and 250 nmol/L). After that, immunofluorescence staining was used to identify differentiated neurons with neuronal specific antibody, Tuj1, and differentiated astrocytes with astrocyte specific antibody, GFAP. Then, we calculated and statistically analyzed Tuj1+/DAPI and GFAP+/DAPI percentages in the three different groups. Besides, real-time PCR assay was used to test Tuj1 and GFAP mRNA expression in the three groups after 3 days of differentiation. RESULTS AND CONCLUSION: Primary and passaged NSCs were successfully cultured and almost of cells were positive for both Nestin and Sox2, so these high-purity NSCs could be used in the following experiments. Immunofluorescence staining and statistical analysis results showed that compared with the blank control and 250 nmol/L groups, 50 nmol/L group had an obviously increased neuronal percentage after 3 days differentiation (P < 0.01), while the blank control and 250 nmol/L groups had no significant difference (P > 0.05); compared with the blank control group, 50 and 250 nmol/L groups had significantly increased astrocyte percentages (P < 0.05), whereas there was no obvious difference between 50 and 250 nmol/L groups (P > 0.05). The results of real-time PCR assay were similar with the above immunofluorescence results. In conclusion, 50 nmol/L ginsenoside Rg3 can enhance mouse NSCs differentiation into neurons and astrocytes, while 250 nmol/L ginsenoside Rg3 can only promote mouse NSCs differentiation into astrocytes.

Identification of the difference in the pathogenesis in heart failure arising from different etiologies using a microarray dataset

OBJECTIVES: Clinically, patients with chronic heart failure arising from different etiologies receive the same treatment. However, the prognoses of these patients differ. The purpose of this study was to elucidate whether the pathogenesis of heart failure arising from different etiologies differs. METHODS: Heart failure-related dataset GSE1145 was obtained from the Gene Expression Omnibus database. Differentially expressed genes were identified using R. A protein-protein interaction network of the differentially expressed genes was constructed using Search Tool for the Retrieval of Interacting Genes. The modules in each network were analyzed by Molecular Complex Detection of Cytoscape. The Database for Annotation, Visualization and Integrated Discovery was used to obtain the functions of the modules. RESULTS: Samples contained in GSE1145 were myocardial tissues from patients with dilated cardiomyopathy, familial cardiomyopathy, hypertrophic cardiomyopathy, ischemic cardiomyopathy, and post-partum cardiomyopathy. The differentially expressed genes, modules, and functions of the modules associated with different etiologies varied. Abnormal formation of extracellular matrix was overlapping among five etiologies. The change in cytoskeleton organization was specifically detected in dilated cardiomyopathy. The activation of the Wnt receptor signaling pathway was limited to hypertrophic cardiomyopathy. The change in nucleosome and chromatin assembly was associated with only familial cardiomyopathy. Germ cell migration and disrupted cellular calcium ion homeostasis were solely detected in ischemic cardiomyopathy. The change in the metabolic process of glucose and triglyceride was detected in only post-partum cardiomyopathy. CONCLUSION: These results indicate that the pathogenesis of heart failure arising from different etiologies varies, which may provide molecular evidence supporting etiology-based treatment for heart failure patients.

Studies on flavonoids from Cirsium setosum / 中国中药杂志

To investigate the chemical constituents of ethyl acetate from Cirsium setosum, fifteen flavonoids were obtained by column chromatography on silica gel, MCI, Sephadex LH-20, and preparative HPLC. Their structures were identified as 4',5,6-trihydroxy-7-methoxyflavone(1), 4',5-dihydroxy-7,8-dimethoxyflavone(2), sorbifolin-6-O-β-glucopyranoside(3), kaempferol-7-O-α-L-rhamnoside(4), kaempferol(5), quercetin-3-O-β-D-glucosyl-7-O-α-L-rhamnoside(6), myricetin(7), myricetin-3-O-β-D-glucoside(8), 5,7- dihydroxy -3',4'- dimethoxyflavone(9), 3',4',5- trihydroxy-3,7-dimethoxyflavone(10), 3',3,4',5-tetrahydroxy-7-methoxyflavone(11), 3'-hydroxy-4',5,7-trimethoxyflavone(12), 7-hydroxy-3',4',5-trimethoxyflavone(13), 4',5-dihydroxy-2',3',7,8-tetramethoxylflavone(14), and 5-hydroxy-2',3',7,8-tetramethoxylflavone(15) by spectroscopic data analysis. All compounds were isolated from this plant for the first time.Compounds(1-15) were evaluated for their hypoglycemic activities by PTP1B enzyme model. Among them, compounds 2, 12, and 14 showed significant PTP1B inhibitory activities with IC₅₀ values of 2.54, 1.85, 2.11 μmol•L⁻¹, respectively.

Mechanism of low molecular weight GTP binding protein RAC1 in injury of neural function of rats with cerebral ischemia reperfusion

OBJECTIVE@#To discuss the mechanism of low molecular weight GTP binding protein RAC1 in the injury of neural function based on building the rat model of cerebral ischemia reperfusion.@*METHODS@#Middle cerebral artery of rats was ligated and the ligature was released to restore the perfusion after 2 h, the rat model of cerebral ischemia reperfusion injury was built, while the middle cerebral artery was ligated. The rats were randomly divided into the sham group, cerebral ischemia reperfusion group (I/R group) and the group with the injection of RAC1 activity inhibitor NSC23766 (NSC group). The survival and neurological severity score of rats in each group were observed and recorded. Nissl staining was employed to observe the nerve cells, and Western blot to detect expression of RAC1, superoxide dismutase and malondialdehyde.@*RESULTS@#Number of nerve cells for rats in NSC group was significantly more than that in I/R group, but significantly less than that in sham group, with the statistical difference (P < 0.05). The brain water content for rats in NSC group was significantly lower than that in I/R group, but significantly higher than that in sham group, with the statistical difference (P < 0.05). The expression of RAC1 and malondialdehyde for rats in NSC group was significantly lower than that in I/R group, but higher than that in sham group; while the expression of superoxide dismutase was lower than that in sham group, but higher than that in I/R group, with the statistical difference (P < 0.05).@*CONCLUSIONS@#The inhibition of RAC1 activity can reduce the oxidative stress, reduce the neurologic impairment because of cerebral ischemia reperfusion and thus protect the neural function.

Mechanism of low molecular weight GTP binding protein RAC1 in injury of neural function of rats with cerebral ischemia reperfusion

Objective To discuss the mechanism of low molecular weight GTP binding protein RAC1 in the injury of neural function based on building the rat model of cerebral ischemia reperfusion. Methods Middle cerebral artery of rats was ligated and the ligature was released to restore the perfusion after 2 h, the rat model of cerebral ischemia reperfusion injury was built, while the middle cerebral artery was ligated. The rats were randomly divided into the sham group, cerebral ischemia reperfusion group (I/R group) and the group with the injection of RAC1 activity inhibitor NSC23766 (NSC group). The survival and neurological severity score of rats in each group were observed and recorded. Nissl staining was employed to observe the nerve cells, and Western blot to detect expression of RAC1, superoxide dismutase and malondialdehyde. Results Number of nerve cells for rats in NSC group was significantly more than that in I/R group, but significantly less than that in sham group, with the statistical difference (P < 0.05). The brain water content for rats in NSC group was significantly lower than that in I/R group, but significantly higher than that in sham group, with the statistical difference (P < 0.05). The expression of RAC1 and malondialdehyde for rats in NSC group was significantly lower than that in I/R group, but higher than that in sham group; while the expression of superoxide dismutase was lower than that in sham group, but higher than that in I/R group, with the statistical difference (P < 0.05). Conclusions The inhibition of RAC1 activity can reduce the oxidative stress, reduce the neurologic impairment because of cerebral ischemia reperfusion and thus protect the neural function.

Subdivision of M category for nasopharyngeal carcinoma with synchronous metastasis: time to expand the M categorization system / 癌症

<p><b>INTRODUCTION</b>The current metastatic category (M) of nasopharyngeal carcinoma (NPC) is a "catch-all" classification, covering a heterogeneous group of tumors ranging from potentially curable to incurable. The aim of this study was to design an M categorization system that could be applied in planning the treatment of NPC with synchronous metastasis.</p><p><b>METHODS</b>A total of 505 NPC patients diagnosed with synchronous metastasis at Sun Yat-sen University Cancer Center between 2000 and 2009 were involved. The associations of clinical variables, metastatic features, and a proposed M categorization system with overall survival (OS) were determined by using Cox regression model.</p><p><b>RESULTS</b>Multivariate analysis showed that Union for International Cancer Control (UICC) N category (N1-3/N0), number of metastatic lesions (multiple/single), liver involvement (yes/no), radiotherapy to primary tumor (yes/no), and cycles of chemotherapy (>4/≤4) were independent prognostic factors for OS. We defined the following subcategories based on liver involvement and the number of metastatic lesions: M1a, single lesion confined to an isolated organ or location except the liver; M1b, single lesion in the liver and/or multiple lesions in any organs or locations except the liver; and M1c, multiple lesions in the liver. Of the 505 cases, 74 (14.7%) were classified as M1a, 296 (58.6%) as M1b, 134 (26.5%) as M1c, and 1 was not specified. The three M1 subcategories showed significant difference in OS [M1b vs. M1a, hazard ratio (HR) = 1.69, 95% confidence interval (CI) = 1.16-2.48, P = 0.007; M1c vs. M1a, HR = 2.64, 95% CI = 1.75-3.98, P < 0.001].</p><p><b>CONCLUSIONS</b>We developed an M categorization system based on the independent factors related to the prognosis of patients with metastatic NPC. This system may be helpful to further optimize individualized care for NPC patients.</p>

In vitro anti-inflammatory and free radical scavenging activities of flavans from Ilex centrochinensis / 中国中药杂志

This study was carried out to evaluate the anti-inflammatory and free radical scavenging activities of flavans from flex centrochinensis S. Y. Hu in vitro and their structure-activity relationship. LPS-stimulated RAW 264.7 macrophage was used as inflammatory model. MTT assay for cell availability, Griess reaction for nitric oxide (NO) production, the content of TNF-alpha, IL-1beta, IL-6 and PGE, were detected with ELISA kits; DPPH, superoxide anion and hydroxyl free radicals scavenging activities were also investigated. According to the result, all flavans tested exhibited anti-inflammatory effect in different levels. Among them, compounds 1, 3, 4 and 6 showed potent anti-inflammatory effect through the inhibition of NO, TNF-alpha, IL-lp and IL-6, of which 1 was the most effective inhibitor, however, 2 and 5 were relatively weak or inactive. The order of free radical scavenging activities was similar to that of anti-inflammatory activities. Therefore, these results suggest that 3, 4 and 6, especially of 1, were,in part responsible for the anti-inflammatory and free radical scavenging activity of Ilex centrochinensis. Hydroxyl group at 4'-position of B-ring plays an important role in the anti-inflammatory and free radical scavenging capacities.

Effect of astaxanthin on oxidative stress of red blood cells and peroxidation damage of membrane / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of astaxanthin (ASTA) on oxidative stress of intra- and extra- red blood cells during stored period and the protective function for cell membrane.</p><p><b>METHODS</b>The blood of volunteers was collected to prepare suspended red blood cells without leukocytes. Then the red blood cells were randomly divided into group A, group B, group C and group D. The ASTA was added into MAP preservation solution of group B, group C and group D, the final concentration of ASTA was 5, 10 and 20 µmol/L respectively. Group A was used as control group, in which only the dissolved liquid DMSO of ASTA was added. The red blood cells were stored in refrigerator at 2 °C-6 °C. On day 7, 14, 28 and day 42 of storage, the content of reactive oxygen species (ROS) in red blood cells was detected by fluorescence microplate reader. The content of malondialdehyde (MDA) was detected with TBA method. The content of hydrogen peroxide (H2O2) outside cell was detected with spectrophotometric method. The mean corpuscular volume(MCV) was detected with blood cell analyzer. The content of free hemoglobin(FHb) was detected with chemical colorimetry.</p><p><b>RESULTS</b>The ROS, MDA, FHb and H2O2 levels in B, C and D groups were lower than those in control group during the stored period. On day 7 and 14 of storage, among group B, group C, group D and group A, the MCV showed no difference in comparison with control group. On day 28 and 42 of storage, the MCV in B, C and D groups was lower than that in control group.</p><p><b>CONCLUSION</b>The ASTA can reduce the oxidative stress level of stored red blood cells inside and outside, relieve the peroxidation damage of cell membrane.</p>