Chemical constituents in ethyl acetate and butanol fractions of ethanol extracts from Acorus tatarinowii were separated by column chromatography. Bufo skeletal muscle fatigue model was established to study the anti-fatigue activity of separated compounds. Five compounds were separated and identified by spectroscopic analysis as acoramone(1),cycloartenone(2),2,4,5-trimethoxyl-2'-butoxy-1,2-phenyl propandiol(3),5-hydroxymethyl furfural(4), and 5-butoxymethyl furfural(5). Compound 3 was a new compound, and compounds 2 and 5 were separated from this plant for the first time. Compound 4 exhibited a notable anti-fatigue activity.
Acorus ; chemistry ; Animals ; Bufonidae ; Fatigue ; drug therapy ; Muscle, Skeletal ; drug effects ; Plant Extracts ; chemistry ; pharmacology

OBJECTIVETo analyze clinical characteristics, genetic mutation and therapeutic effect of seven patients diagnosed with congenital hyperinsulinism(CHI).

METHODSClinical data for the patients was retrospectively analyzed.

RESULTSAll patients presented with hyperinsulinism(serum insulin:2.0-58.4 mU/L),even after hypoglycemia (blood glucose: 0.7-2.39 mmol/L) has developed. Mutations were identified in 4 patients (57.1%), which included a heterozygous c.262C to T(p.R88C) mutation in exon 4 of the UCP2 gene, a heterozygous c.1495C to A(p.G499C) mutation in exon 12 of the GLUD1 gene, a heterozygous c.1493C to T(p.S498L) mutation in exon 1 of the GLUD1 gene, and a heterozygous c.4432G to A(p.G1478R) mutation in exon 37 of the ABCC8 gene. The patient carrying a maternally inherited ABCC8 mutation was treated with cornstarch and had his blood glucose kept normal. All other patients responded well to diazoxide.

CONCLUSIONA genetic diagnosis was attained for 51.7% of patients in this study. Mild CHI patients can have their blood glucose controlled by giving cornstarch. Diazoxide is safe and effective for most CHI patients.

OBJECTIVE: To summarize clinical manifestations, inheritance pattern and mutations of NR0B1 gene in 7 children with X-linked adrenal dysplasia congenita (XL-AHC). METHODS: Clinical data of the 7 children was collected. Next-generation sequencing was carried out to detect potential mutations in the coding regions of adrenal gland-related genes. Suspected mutations were verified with Sanger sequencing. RESULTS: In all of the children, the initial symptom was adrenocortical insufficiency. Five cases had neonatal onset, while the remaining two developed it at the age of 2. Three cases (42.9%) had a short stature and 1 showed growth retardation (14.3%). Of the 7 cases, 6 (85.7%) had mutations occurring in exon 1, and 1 (14.3%) had it occurring in exon 2. Four cases (57.1%) were frameshift mutations, 2 cases (28.6%) were nonsense mutations and 1 case (14.3%) was missense mutation. Two mutations were known to be pathogenic, and 5 had not been reported previously. Maternal inheritance was found in 6 cases. Three children had a maternal uncle died of unexplained causes. The mothers of 2 children had a history of spontaneous abortions. One child had a brother died of unexplained reason. CONCLUSION Male children with primary adrenal insufficiency should be routinely checked for NR0B1 mutations, especially those with a family history. mutations of NR0B1 gene occur mostly in exon 1, with frameshift mutations being the most common type. The development of all patients with XL-AHC should be closely monitored during follow-up.
Adrenal Insufficiency ; Child ; DAX-1 Orphan Nuclear Receptor ; DNA Mutational Analysis ; Genes, X-Linked ; Humans ; Hypoadrenocorticism, Familial ; Male ; Mutation

Toxoplasma gondii is an obligate intracellular parasite that stimulates production of high levels of proinflammatory cytokines, which are important for innate immunity. NLRs, i.e., nucleotide-binding oligomerization domain (NOD)-like receptors, play a crucial role as innate immune sensors and form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1β. To elucidate the role of inflammasome components in T. gondii-infected THP-1 macrophages, we examined inflammasome-related gene expression and mechanisms of inflammasome-regulated cytokine IL-1β secretion. The results revealed a significant upregulation of IL-1β after T. gondii infection. T. gondii infection also upregulated the expression of inflammasome sensors, including NLRP1, NLRP3, NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and NAIP, in a time-dependent manner. The infection also upregulated inflammasome adaptor protein ASC and caspase-1 mRNA levels. From this study, we newly found that T. gondii infection regulates NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and neuronal apoptosis inhibitor protein (NAIP) gene expressions in THP-1 macrophages and that the role of the inflammasome-related genes may be critical for mediating the innate immune responses to T. gondii infection.
Apoptosis ; Cytokines ; Gene Expression ; Immunity, Innate ; Inflammasomes* ; Macrophages* ; Multiprotein Complexes ; Negotiating ; Neurons ; Parasites ; RNA, Messenger ; Toxoplasma* ; Up-Regulation

OBJECTIVETo study the expression of vascular endothelial growth factor (VEGF) and metastin in colorectal carcinoma and their association with the clinicopathological features of the malignancy.

METHODSVEGF and metastin expressions were examined immunohistochemically with SP method in 70 specimens of human colorectal carcinoma tissues and adjacent normal tissues.

RESULTSVEGF protein overexpression was detected in 48.6% (34/70)of the colorectal carcinoma tissues but in none of the adjacent normal tissues (P<0.01), and for metastin, the overexpression rate was 28.6% (20/70) in the colorectal carcinoma tissues and 70.0% (49/70) in the normal tissues (P<0.01). The expression of both VEGF and metastin was related to the histological grades, infiltration depth, TNM stage and lymph node metastasis of the tumor (P<0.01 and P<0.05, respectively).

CONCLUSIONImmunohistochemical detection of VEGF and metastin can be of value in assessment of the malignancy and in prognostic evaluation of colorectal carcinoma.

Adult ; Aged ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kisspeptins ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Tumor Suppressor Proteins ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Young Adult

OBJECTIVEThrough maternal inheritance, to explore the genetic structures and relationships of Dong, Gelao, Tujia and Yi ethnic population in Guizhou of China.

METHODSThe mtDNA D-loop hypervariable segment I (HVS I ) in 108 samples of four ethnic populations were sequenced. Then, the nucleotide diversity was estimated and a phylogenetic tree was constructed by Neighbor-Joining method.

RESULTSIn the detected 497 bp fragments, 86 polymorphic sites were found, and 82 different haplotypes were identified. The phylogenetic tree of four ethnic populations showed: Yi, Tujia and Gelao clustered more closely than Dong did.

CONCLUSIONYi and Tujia population are very closely related, the reason may be that they either originate from a common ancestry or frequently undergo the gene exchanges and admixtures. The genetic relationship between Tujia and Gelao population is nearer, perhaps because they have settled in the adjacent regions. Dong and Yi population show the farthest genetic relationship, this is probably due to their different historical origins and geographic segregation.

Base Sequence ; China ; DNA, Mitochondrial ; chemistry ; classification ; genetics ; Ethnic Groups ; genetics ; Genetic Variation ; Humans ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo detect changes of serum soluble Apo-1/Fas (sApo-1/Fas) in pancreatic cancer patients and to investigate its clinical value in assessing the effect of chemotherapy.

METHODSThe serum level of sApo-1/Fas in 30 normal control subjects and 58 pancreatic cancer patients were detected using enzyme-linked immunosorbent assay (ELISA), and the sApo-1/Fas level of 48 pancreatic cancer patients, before and after chemotherapy was compared.

RESULTSCompared with the level of the control group, the level of serum soluble Apo-1/Fas was significantly correlated with clinical stage but not with age, sex or pathologic type of pancreatic cancer. It was elevated gradually from stage II to IV (P < 0.01). However, it would obviously decrease in pancreatic cancer patients after chemotherapy (P < 0.01).

CONCLUSIONThe serum soluble Apo-1/Fas may be involved in the development of pancreatic cancer, and it may be used as one parameter to assess the disease status and prognosis of pancreatic cancer patient.

Adenocarcinoma, Mucinous ; blood ; drug therapy ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Pancreatic Ductal ; blood ; drug therapy ; Cisplatin ; administration & dosage ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; blood ; drug therapy ; Prognosis ; Remission Induction ; fas Receptor ; blood

Toxoplasma gondii is an intracellular parasite that causes severe disease when the infection occurs during pregnancy. Adenosine is a purine nucleoside involved in numerous physiological processes; however, the role of adenosine receptors in T. gondii-induced trophoblast cell function has not been investigated until now. The goal of the present study was to evaluate the intracellular signaling pathways regulated by adenosine receptors using a HTR-8/SVneo trophoblast cell model of T. gondii infection. HTR8/SVneo human extravillous trophoblast cells were infected with or without T. gondii and then evaluated for cell morphology, intracellular proliferation of the parasite, adenosine receptor expression, TNF-α production and mitogen-activated protein (MAP) kinase signaling pathways triggered by adenosine A3 receptor (A3AR). HTR8/SVneo cells infected with T. gondii exhibited an altered cytoskeletal changes, an increased infection rate and reduced viability in an infection time-dependent manner. T. gondii significantly promoted increased TNF-α production, A3AR protein levels and p38, ERK1/2 and JNK phosphorylation compared to those observed in uninfected control cells. Moreover, the inhibition of A3AR by A3AR siRNA transfection apparently suppressed the T. gondii infection-mediated upregulation of TNF-α, A3AR production and MAPK activation. In addition, T. gondii-promoted TNF-α secretion was dramatically attenuated by pretreatment with PD098059 or SP600125. These results indicate that A3AR-mediated activation of ERK1/2 and JNK positively regulates TNF-α secretion in T. gondii-infected HTR8/SVneo cells.

Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), IFN-γ, TNF-α, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.
Animals ; DNA ; Humans ; Immunity, Cellular ; Immunization ; Immunoglobulin G ; Incidence ; Interleukin-10 ; Interleukin-12 ; Mice ; Parasites ; Plasmids ; Toxoplasma ; Toxoplasmosis ; Vaccination