This study was aimed to reveal the effects and molecular regulation mechanism of methyl jasmonate (Me-JA) on volatile terpenoids from Amomum villosum Lour. After the leaves and fruits of A momum villosum Lour. were treated with different concentrations of MeJA, the volatile terpenoids of fresh fruits from A . villosum Lour. were ex-tracted with microwave method and analyzed by GC-MS. Then, leaves and fruits treated with MeJA were sequenced by Illumina. The transcriptome data was analyzed by bioinformatic methods. The results showed that there were 20 and 33 volatile terpenoids detected in peels and seed groups, respectively. Contents of volatile terpenoids in peels and seed groups were both improved after 600 μmol·L-1 MeJA treating fruits for 24 h, such as bornyl acetate, cam-phor, borneol, and etc. While 200 μmol·L-1 MeJA treating different parts for 24 h can regulate the biosynthesis of some volatile terpenoids in peels differently. And 200 μmol·L-1 MeJA treating fruits can improve the content of ma-jor volatile terpenoids in seed groups. A total of 68 168 unigenes were obtained with de novo assembly, and 48 627 unigenes were annotated after comparison with public protein databases. Analysis of functional annotation against KEGG database showed that there were 208 unigenes closely related with metabolism of volatile terpenoids and 22 u-nigenes related with MYC2 transcription factors. It was concluded that MeJA can effectively regulate the metabolism of volatile terpenoids from A . villosum Lour. There were a lot of candidate genes related with the biosynthesis of volatile terpenoids obtained by analyzing the transcriptome data which also provided a large amount of data for the discovery and regulation of functional genes related with the biosynthesis of volatile terpenoids from A . villosum Lour.

Objective To identify the reliable reference genes for gene expression analysis of the pericarp and seed of Amomum villosum Lour. by using real-time fluorescence quantitative polymerase chain reaction ( qRT-PCR). Methods Using the fruits ( separated into peels and seeds) of A. villosum at three different developmental periods as the experimental material, 5 candidate reference genes (β-actin, EF-1α, GAPDH, PGK, TUA) with steady expression were screened out by the high throughout sequencing of transcriptome and expression profile data. The qRT-PCR technique was applied to study the expression levels of 5 candidate reference genes in different samples. The stability of the candidate reference genes were evaluated by GeNorm and NormFinder software. Results The 5 reference genes had different stabilities in the pericarp and seed of A. villosum Lour. at different development periods . The order of the steadiness of reference genes showed by GeNorm was EF-1α = TUA>PGK>GAPDH>β-actin. The results of NormFinder revealed that EF-1α was the most stable, followed by TUA, and the order of the other three genes was as same as the results of GeNorm. Conclusion EF-1αand TUA could be used as double reference genes for the normalization of gene expression in A. villosum fruits at different developmental periods by using qRT-PCR.

Objective To investigate the variance of cost-effectiveness when treat acute myocardial infaretion of different severe extents with different pattern.Methods Acute myocardial infarction patients were selected from emergency eommand center of Guangzhou from October 2003 to December 2005.These patients wew assigned by the center to First-Class Hospitals at Grade 3 and First-class Hospital at Grade 2,and were followed up after 6 months after post-discharge.Cost in hospital and mortality in hospital were registered.The health of all patients were quantificated using SF-36.According to the assigned hospitals,the patients were divided into single infarction group and complex infarction group.Cost in hospital,mortality in hospital,short-term quality of life were compared between the them.Results Compared with and First-Class Hospital at Grade 2 (101 cases),the single infarction patients in First-Class Hospitals at Grade 3 had higber costs in hospital (P=0.016),better society function,affection role,mental health and health status (P

Objective:To investigate the effects and the possible mechanisms of Neiguan(PC6)and Gongsun(SP4)on the hypothalamic-pituitary-adrenal(HPA)axis in rats with functional dyspepsia(FD),thus to provide a theoretical basis for the clinical application of the Eight Confluent Points.Methods:Forty specific-pathogen-free Sprague-Dawley rats were divided into a blank group,a model group,an electroacupuncture(EA)group,and a Western medicine group by the random number table method,with 10 rats in each group.Rats in the blank group did not receive modeling or intervention.Rats in the other three groups were subjected to the FD with mood disorder model using the compound etiology modeling method.After the successful modeling,rats in the model group did not receive any interventions,rats in the Western medicine group received deanxit and mosaprid intervention,and those in the EA group received EA intervention on the ipsilateral Neiguan(PC6)and Gongsun(SP4)for 21 d.The sugar-water consumption rate was measured before the experiment and before and after interventions to assess the emotional status.The gastric emptying rate was measured after interventions to assess the gastrointestinal dynamics.The expression levels of hypothalamic corticotropin-releasing hormone(CRH),pituitary adrenocorticotropic hormone(ACTH),and adrenal corticosterone(CORT)were measured by enzyme-linked immunosorbent assay.Results:Compared with the blank group,the sugar-water consumption rate and the gastric emptying rate were decreased(P<0.01),and the hypothalamic CRH,pituitary ACTH,and adrenal CORT expression levels were increased(P<0.01)in the model group.Compared with the model group,the sugar-water consumption rate and the gastric emptying rate were significantly increased(P<0.01),while the expression levels of hypothalamic CRH,pituitary ACTH,and adrenal CORT were significantly decreased(P<0.01)in the EA group and the Western medicine group.The differences between the EA group and the Western medicine group were not statistically significant(P>0.05).Conclusion:The Eight Confluent Points Neiguan(PC6)and Gongsun(SP4)can improve the mood and gastrointestinal dynamics in FD rats,which may be achieved by down-regulating the hypothalamic CRH,pituitary ACTH,and adrenal CORT,as well as by correcting the HPA axis hyperfunction.

OBJECTIVETo investigate the intervention and mechanism of ambroxol combined with low-dose heparin on oxidative stress, TNF-alpha and IL-1beta in rabbits with acute lung injury (ALI).

METHODSTwenty-four healthy Japanese rabbits were randomly divided into three groups: (1) Normal saline control group (NC), (2) Oleic acid injury group (OA), (3) Ambroxol + low-dose heparin therapy group (AH). After the success of ALI model, AH group was injected ambroxol + low-dose heparin, while the NC group and OA group were injected the same dose of normal saline by the same method. Arterial oxygen tension (PaO2), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) at different time points were determined. The pathological manifestation of both side lungs was observed at the end of expeiment. The activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), xanthine oxidase (XO) and the content of malondialdehyde (MDA) in bronchoalveolar lavage fluid (BALF) and lung tissue homogenate were tested. The apoptosis index was detected. The lung wet/dry (W/D) ratio was calculated. The pathological changes in lung tissue were observed by light microscopy, and the ultrastructural changes of lung tissue were observed by electron microscopy.

RESULTS(1) The instructive injury induced by ALI observed under electron microscope and light microscope and W/D was decreased significantly in AH group. (2) PaO2 was improved significantly in AH group, compared with that in OA group (P < 0.01). (3) The activity of GSH-Px and SOD in AH group increased significantly (P < 0.01 or P < 0.05) but the activity of XO and the content of MDA decreased significantly (P < 0.01), compared with those in OA group. (4) Except the content of IL-1beta in serum before treatment, the content of IL-1beta and TNF-alpha in serum, BALF, lung tissue homogenate of OA group increased significantly (P < 0.01), and those were obviously improved in AH group. (5) Apoptosis index (AI) in AH group decreased significantly (P < 0.01) compared with that in OA group.

CONCLUSIONIn ALI induced by OA, IL-1beta and TNF-alpha increases significantly and involved in the occurrence and development of ALI. Ambroxol combined with low-dose heparin can reduce lung cells oxidative stress to inhibit the release of IL-1beta and TNF-alpha, which play a role in the treatment of ALI.

Acute Lung Injury ; chemically induced ; drug therapy ; metabolism ; Ambroxol ; therapeutic use ; Animals ; Drug Therapy, Combination ; Female ; Heparin ; administration & dosage ; Interleukin-1beta ; metabolism ; Male ; Oleic Acids ; Oxidative Stress ; drug effects ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Adjuvants, Immunologic ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

Objective To investigate the role of c-Jun NH2-terminal kinase (c-JNK) signaling pathway on voltage-gated potassium channel (Kv) remodeling in left ventricular myocytes of diabetic rats,and explore the intrinsic regulatory mechanism.Methods Forty-five SD rats were randomly divided into DM group (n=25,modeling with streptozotocin induction) and control group (n=20,fed with normal diet).Transient outward potassium current (Ito) of rats' ventricular myocytes in DM group and control group was recorded by whole-cell patch-clamp method.The c-Jun activity was detected using a non-radioactive JNK kinase assay kit (Cell Signaling Technology).JNK inhibitor SP600125 was used to incubate the cardiomyocytes of diabetes rats in vitro,and then the changes of I,o in cardiomyocytes were observed.Thioredoxin reductase (TrxR) inhibitor--auranofin (AF) was used to treat the rats' cardiomyocytes incubated with SP600125,and then the changes of Ito in cardiomyocytes were observed.The content of Kv4.2 was tested using anti-Kv4.2 antibody,and the results were analyzed using a UVP bioimaging system.Results The JNK activity in DM group rose more than 1 times compared with control group,while the density of Ito decreased significantly (Control:30.2 ± 3.3pA/pF,n=16;DM:15.3 ± 2.1pA/pF,n=17;P＜0.05).The ventricular myocytes of DM rats were treated with SP600125 (10μmol/Lol/L) for 4 hours,then the Ito density increased to control group level (DM+SP600125:32.3 ± 3.7pA/pF,n=18;Control:30.2 ± 3.3pA/pF,n=16;P＜0.05).There was no significant difference in the maximum Ito density between the treated with SP600125 (Control+SP600125:31.6 ± 3.4pA/pF,n=18) and untreated control groups.The Ito density in DM myocardial cells significantly increased after treatment with the membrane permeable protein inhibitor JNKI-1 (10μmol/L),and no changes were found in control group after the same treatment.The augmentation effect of SP600125 on Ito current in DM myocytes was significantly inhibited by TrxR inhibitor auranofin (lμmol/L) (DM+SP600125+AF:15.7 ± 3.3pA/pF,n=15),while AF did not change the Ito density in control group.The expression of Kv4.2 protein was significantly increased in DM rats after administration of SP600125,which was consistent with the changes of Ito current observed in the myocardium of DM rats,although not fully restored to the level of control group myocardium.JNK inhibitor did not markedly alter the expression of Kv4.2 protein in control group myocardium.Conclusions Kv channel remodeling in DM rat's myocardium is redox-regulated,and the Ito remodeling might be assisted with the persistent activation of c-JNK signaling pathway.It has showed that c-JNK activity is significantly increased in DM rat heart and the current density of Kv channels is reduced.The inhibition of JNK signaling pathway can markedly improve Kv channel reconstruction and the process may be regulated by thioredoxin system.

Purpose/Significance To elucidate the complex relationship among face phenotype-disease-gene,and to explore the underlying mechanisms of their interactions.Method/Process Based on scientific literature from the PubMed database,natural language processing tools and manual filtering methods are used to extract the knowledge of the concept and relationship of face phenotype-disease-gene reported in the existing literature,and a knowledge base is constructed.Result/Conclusion The study completes the framework design and construction of the knowledge base of the face phenotype-disease-genotype for intelligent health application,which lays a foundation of both data and theory for exploring the interplay between face phenotype-disease-gene,as well as the potential application of face phenotype in disease diagnosis.

Extranodal natural killer/T-cell lymphoma (ENKTCL) is the subtype of non-Hodgkin lymphoma with high heterogeneity and invasiveness. Though most ENKTCL patients are present as early-stage at diagnosis, clinical prognosis significantly differs due to the limitations of clinical staging. Radiotherapy (RT) and chemotherapy (CT) are the first-line treatments for early ENKTCL patients. However, there is no consensus on the combined modalities of RT and CT, and their optimal strategy. With the continuous renewal of clinical staging and prognostic models, early-stage ENKTCL patients tend to accept risk-adapted treatment with proper stratification. In this review, the latest research progresses on clinical staging, prognostic models and treatment were retrospectively analyzed, aiming to provide references for clinical decision-making.

BACKGROUND: We previously found that the intestinal epithelial chemokine (C-C motif) ligand 7 (CCL7) plays an important role in the development of toxin-induced acute liver damage. The detailed effects of intestinal epithelial CCL7 on chronic diseases; however, are still unclear. Here, we aimed to investigate the impact of intestinal epithelial CCL7 overexpression on high-fat diet (HFD)-induced obesity and steatohepatitis in mice. METHODS: Intestinal epithelial CCL7 overexpression (CCL7) mice and their wild-type (WT) littermates were fed with normal chow or HFD for 16 weeks to induce obesity and non-alcoholic fatty liver disease. Body weight gain, as well as adipose tissue index were assessed. Liver injury was monitored by histological analysis and real time polymerase chain reaction. Gut microbial composition was analyzed by 16S rRNA gene sequencing. RESULTS: We found that the CCL7 mice on a HFD had markedly decreased weight gain (8.9 vs. 17.0 g, P < 0.05) and a lower adipose tissue index that include mesenteric fat (1.0% vs. 1.76%, P < 0.05), gonadal fat (2.1% vs. 6.1%, P < 0.05), subcutaneous fat (1.0% vs. 2.8%, P < 0.05) compared to WT animals. HFD-induced glucose intolerance and insulin resistance were also significantly improved in CCL7 mice compared to WT. Furthermore, HFD-fed CCL7 mice displayed less hepatic lipid accumulation and lower expression of inflammatory factors than WT mice. 16S rRNA gene sequencing demonstrated that CCL7 overexpression in intestinal epithelial cells improved HFD-induced gut microbial dysbiosis. CONCLUSIONS Our study revealed that CCL7 overexpression in the intestinal epithelium protects mice against the progression of diet-induced obesity, hepatic steatosis, and enteric dysbiosis.