Objective To find the relationship between mutation of gamma 2 subunit of the gamma-aminobatyric acid type A receptor(GABRG2) and generalized epilepsy with febrile seizure plus(GEFS+).Methods Probands of 10 families with GEFS+ were selected,the GABRG2 gene were sequenced.Results We found a single nucleotide polymorphism site,and did not find the reported mutations.Conclusion GABRG2 mutation is not common in Hans of northern China.

OBJECTIVETo explore the specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DC), which were transfected by the plasmid pC53-SN3 encoding p53 gene.

METHODSDC derived from HLA-A2(+) mononuclear cells of the 24-lung cancer patients was transfected with the plasmid pC53-SN3 by lipofectamine and then co-cultured with auto-unpurified T cells to induce potent CTL (T-pC53-SN3). The cytolysis of specific CTL against Calu-6, a HLA-A2(+) human lung cancer cell line, was measured by using lactate dehydrogenase (LDH) releasing assay.

RESULTSThe expression of CD(1a) and CD(83), the correlative markers of DC, increased apparently after transfected with plasmid pC53-SN3, the expression rate was (5.45 +/- 0.89)% and (3.26 +/- 0.47)% versus (52.15 +/- 11.56)% and (25.78 +/- 12.35)%. CD(14) decreased apparently, but other DC correlative markers of CD(1a), CD(40), CD(86), and HLA-DR remained almost the same as that before transfection. Compared with T-IL-2, the CTL derived from PBMNC stimulated by IL-2 (100 U/ml), the cytolytic activity of T-pC53-SN3 against Calu-6 cell line showed a significant increase, but cytolytic activity was 56.79 +/- 15.67 and 39.33 +/- 9.88, respectively, when effect cells: target cells was 10:1. The expression of the CD(8), CD(69), and CD(45)RO/CD(8) of T-pC53-SN3 cells increased significantly, but that of CD(3), CD(4), CD(86), ect, was not significantly different from those of T-pCMV-neo.

CONCLUSIONSIt showed that DC transfected by p53 gene could induce potent HLA-A(2) restrictive CTL to kill tumor cell efficiently.

Antigens, CD ; analysis ; B7-2 Antigen ; CD40 Antigens ; analysis ; Cell Line, Tumor ; immunology ; Coculture Techniques ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; drug effects ; immunology ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Membrane Glycoproteins ; analysis ; T-Lymphocytes ; immunology ; Tumor Suppressor Protein p53 ; genetics ; physiology

OBJECTIVETo research the effects of Guizhi Fuling capsule on sex hormones levels in blood serum and breast issue morphology of hyperplasia of mammary glands model rats.

METHODThe unpregnancy SD rat models of hyperplasia of mammary glands were established by injecting 0.5 mg x kg(-1) benzoate estradiol. After five weeks doses,the effects of Guizhi Fuling capsule 2.0, 1.0, 0.5 g x kg(-1) and Rupixiao tablet 0.5 g x kg(-1) on the changes of papilla diameter, height and breast issue morphology of the naimal models were explored, and sex hormones levels in blood serum were measured.

RESULTGuizhi Fuling capsule can inhibitnipple swell, improve breast tissue morphology pathological profiles of the animal models, and decrease oestradiol (E2) level and increase progesterone (P) level in blood serum.

CONCLUSIONThese results suggested that Guizhi Fuling capsule could, improve mammary gland pathological profiles. Regulating sex hormone levels may be its important mechanism for treatment of hyperplasia of mammary glands.

Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadal Steroid Hormones ; blood ; Hyperplasia ; Mammary Glands, Animal ; drug effects ; pathology ; Rats

OBJECTIVE: To explore the effects of granulocyte macrophage-colony-stimulating factor (GM-CSF) on the proliferation and activation of dendritic cells (DC) in vivo. METHODS: Balb/c mice were randomly assigned into the control and GM-CSF treatment group. After the lethal dosage of irridiation, green fluorescence protein labelled hematopoietic stem cells (HSC) were injected into the tail veins of the mice. The dosage of 0.1 micorg GM-CSF was administer subcutaneously every 2 days after the HSC infusion. The numbers and activation status of splenic DC were observed by flow cytometry 1, 2, 3, and 4 weeks after the HSC transplantation. RESULTS: The numbers of splenic DC in the GM-CSF treatment group were significantly higher than those of the control group. The expressions of surface marker CD40, CD80, and CD86 in the GM-CSF treatment group were also higher than those of the control group. CONCLUSION GM-CSF can enhance the proliferation and activation of DC in vivo.
Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; Female ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Transplantation ; Male ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Spleen ; cytology ; Whole-Body Irradiation

Background Keratoconus is a bilateral,noninflammatory,gradually progressive corneal disorder characterized by progressive thinning and steepening of the central cornea.It is significant to investigate keratoconusrelated pathogenic gene for elaborating the pathogenesis and establishing early diagnosis standard and taking clinical measurement.Objective The aim of the study was to explore the relationship of visual system homeobox gene (VSX1) polymorphism and the risk of sporadic keratoconus.Methods This study was approved by Ethic Commission of First Hospital of Xi' an.Written informed consent was obtained from each subject prior to enrollment.A case-controlled study was conducted.One hundred and one Han nationality patients with sporadic keratoconus were included in this study.These keratoconus patients were clinically diagnosed by slit lamp examination and corneal tomography.Single nucleolide polymorphism (SNP) of VSX1 gene was assayed and classified using the MassARRAY SNP technique.Demography and relevant risk factors were collected from each subject by questionnaire.Eighty healthy volunteers served as controls.Chi-square test and Binary logistic regression were used to evaluate the difference in the distribution of allele frequency and genotype frequency and to analyze the association with keratoconus risks.Results SNP of two genes was found in the Chinese Han population (rs743018 (c.843+140 C＞T) and rs6138482(R217H C＞T)).There were no significant differences in the genotype frequency and allele frequency of the SNP of two genes in the keratoconus group in comparison with the normal control group (P＞0.05).After adjustment by age and sex,SNP of two genes was not significantly associated with the risk of keratoconus (regression model:rs743018 (C＞T) adjusted:P=0.35,OR=0.72,95％ CI:0.37-1.43 ;rs6138482 (C＞T) adjusted:P =0.48,OR=0.76,95％ CI:0.35-1.64).Conclusions Gene polymorphisms of rs743018(c.843+140 C＞T) and rs6138482(R217H C＞T) in the Chinese Han population is not associated with the risk of keratoconus.Due to the racial difference in genotype and allele frequency,the role of the VSX1 gene in the pathogenesis of keratoconus still remains controversial,and further study needs to be developed.

Encephalomyocarditis virus (EMCV) is a natural epidemic zoonotic pathogen. However, no reports have been published regarding the isolation, identification and full-length genome of EMCV from a local aardvark population. In present study, an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province. The systematic identification, full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13 were conducted. The result showed that the isolate was spherical with a diameter of 24-30 nm, neither heat- nor acid-resistant, sensitive to trypsin, insensitive to chloroform, not protected by bivalent cationic, and the specific fluorescence was observed in the cytoplasm of BHK-21 cells infected with the isolate by using indirect fluorescence assay. The full-length genome of EMCV HNXX13 generated a 7 725bp sequence (GenBank: F771002), with 81.0%-99.9% nucleotide identity to reference strains from different animals, and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd. The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1, G2 and G3, and strain HNXX13 belonging to the G1 group with other Chinese reference strains. The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population, and enriches the molecular epidemiological data of EMCV in China. Regional differences exist in EMCV genome and transmission is limited within a certain area. However, the cross-infection and transmission of EMCV between aardvark and mice appears most likely. Mutations have occurred in some amino acids of EMCV strain HNXX13 during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark.
Animals ; Animals, Wild ; virology ; Cardiovirus Infections ; veterinary ; virology ; China ; Encephalomyocarditis virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Mice ; Molecular Sequence Data ; Phylogeny ; Xenarthra ; virology

OBJECTIVETo explore the regulatory effect of arginine on the secretion of hepatic insulin-like growth factor-1 (IGF-I), and the mechanism of enhancing the immune function by arginine.

METHODSWistar rats were randomly divided into normal control (NC), wound control (WC), and wound with arginine (Arg) groups, with 8 rats in each group. The rats in WC and Arg groups were inflicted with soft tissue trauma on the back. The rats in Arg group were fed a diet supplemented with 5% arginine for one week, while those in NC and WC groups were fed with glycine. The serum contents of arginine, ornithine, growth factor (GH), NO and IGF-I were determined 7 days after feeding. T cell proliferation and IGF-I mRNA expression in hepatic tissue were also measured. Meanwhile, the rat hepatocytes were cultured in serum-free medium containing different concentrations of arginine. The supernatant was collected for the determination of IGF-I level.

RESULTS1). There was no obvious difference of the serum level of arginine and ornithine between NC and WC groups (P > 0.05), but the contents of them were obviously higher in the Arg group compared with other two groups (P < 0.01). 2). No difference in the serum GH level was found among all the groups (P > 0.05), but the serum NO content in WC and Arg groups was significantly lower than that in NC group (P < 0.01), and the serum IGF-I content in WC group decreased obviously compared with that in NC group (P < 0.01). 3). The thymocyte proliferation rate in WC group was also markedly lower than that in NC group (P < 0.01), but that in Arg group was improved compared with WC group (P < 0.01). 4). The expression of hepatic IGF-I mRNA: The relative value of IGF-I mRNA was 1.19 +/- 0.06, 1.08 +/- 0.06 and 1.29 +/- 0.06 in NC, WC and Arg, respectively, while the value in WC was lower than that in NC (P < 0.05) group, and that in Arg group was much higher than that in WC group (P < 0.01). 5). The IGF-I level in the supernatant of cultured hepatocytes: When Arg concentration was 0.0750, 0.7500, 7.5000 mmol/L in the culture medium, the IGF-I level in the supernatant of hepatic cell medi-um was obviously higher than that in the medium without arginine (P < 0.01). Although IGF-I level decreased in the culture medium with arginine in the dose of 37.5000 mmol/L, it was still obviously higher than that in the medium without arginine (P < 0.01).

CONCLUSIONArginine could also produce the immune enhancing effect by stimulating hepatic IGF-I secretion.

Animals ; Arginine ; pharmacology ; Enteral Nutrition ; Insulin-Like Growth Factor I ; metabolism ; Liver ; drug effects ; secretion ; Male ; Rats ; Rats, Wistar ; Soft Tissue Injuries ; metabolism ; therapy

OBJECTIVETo investigate the effects of dendritic cells (DCs) infected with adenovirus vector encoding xenogeneic alpha-fetoprotein (AFP) on breaking the immune tolerance and induction of immunity against hepatocellular carcinomas.

METHODSHuman and mouse alpha-fetoprotein full-length cDNA were cloned from human HepG2 and mouse Hepa 1 - 6 hepatoma cell lines, respectively, using RT-PCR, and then inserted into adenoviral shuttle vectors to construct Ad hAFP and Ad mAFP. Mice were immunized with Ad hAFP-infected DC and in vitro CTL activity against Hepa 1 - 6 cells was examined by standard (51)Cr release assay. Survival was studied of the immunized mice, with or without depletion of CD8+ or CD4+ T cells, inoculated with Hepa 1 - 6 mouse hepatoma cells.

RESULTSThe lytic activity of CTL elicited by the Ad hAFP-infected DCs were much stronger than that by Ad mAFP-infected DCs. 80% of the Ad hAFP/DCs-immunized mice of the inoculated with 5 x 10(6) Hepa 1 - 6 hepatoma cells were still alive two months after inoculation. However, the Ad mAFP/DCs-immunized mice inoculated with 1 x 10(6) Hepa 1 - 6 cells were just 20% surviving two months later. Depletion of CD8+ or CD4+ T cells abolished such an antigen-specific immunity elicited by the DCs infected with Ad hAFP.

CONCLUSIONAdenovirus vector-mediated xenogeneic AFP-infected DCs can effectively break the immune tolerance to hepatocellular carcinomas in an animal model and induce strong antigen-specific T cell response, which are dependent on CD8+ and CD4+ T cells.

Adenoviridae ; genetics ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Female ; Genetic Vectors ; Humans ; Immunization ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; alpha-Fetoproteins ; immunology

OBJECTIVETo isolate, culture and identify a dog periodontal ligament stem cells (PDLSC) line in vitro.

METHODSThe adult dog periodontal ligament cells were isolated by limited dilution of culture cell for single cell clone. Cells originated from one of these clones were assessed through colony-forming efficiency and immunocytochemistry assay and alkaline phosphatase stain was used to identify the source of adult dog periodontal stem cells, at the same time, PDLSC were induced with mineralizatin solution and was found to have long protrude like an osteoblast. Differentiation of PDLSC were assessed. Mineralized potential was studied by Von-Kossa staining.

RESULTSThe dog PDLSC expressed STRO-1, which was the marker of mesenchymal stem cells. Also Vimentin, osteoblast-like marker alkaline phosphatase and Collagen-I expressed weakly. Cells were clonegenic, highly proliferative cells and capable of differentiating into osteoblasts/cementoblasts.

CONCLUSIONThe evidence suggests that the cultured cells were stem cells from adult dog periodontal ligament.

Adult ; Adult Stem Cells ; Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Dental Cementum ; Dogs ; Humans ; Mesenchymal Stromal Cells ; Osteoblasts ; Periodontal Ligament ; Stem Cells

Through researches of channels and collaterals and clinical practice of many years, the authors understand that Chinese medicine, which considers the human body as an interrelated, mutual constraints, whole, dynamic living system, has gradually become an important part of modern medicine. Channels and collaterals are a closed loop system which is communicated and linked by energy and information in the form of electromagnetic oscillation, reflecting many characteristics similar to quantum. Channels and collaterals are not a fixed organizational structure. Studies on channel and collaterals find that the track of the propagated sensation along channels (PSC) have the phenomenon drifting about. This exactly reflects the law of channels dynamically running. By information triggering and living resonance, channels and collaterals bring into play entirely regulative action. The innovative treatment of channels and collaterals followed by characteristics and laws of quantum can get a better curative effect. Theory of channels in the position of quantum information medical science provides an important breach for modernization of Chinese medicine.
Medical Informatics ; Medicine, Chinese Traditional ; trends ; Meridians