Objective: To provide biotechnology reference for the utilization of Epimedium wushanense. Methods: Some subculture conditions were determined and the growth characteristics of suspension cells were observed every 3 d. Results: The cell showed S-shaped growth curve and the content of nitrate, ammonia salt, phosphate, and sucrose all showed sharply decreasing trend during the initial cell suspension culture of E. wushanense. Then the respective changes slowed down. The optimum illumination duration was 12 h. The highest output of flavonoids was 27.854 82 mg/L under 12 h illumination. Conclusion: The growth characteristics research lays the foundation of mass natural active ingredient in cell suspension culture production of E. wushanense.

Adaptive responses to diverse microbial pathogens might be limited in relatively few types. Host cell responses to pathogens are believed to be patterned or stereotyped along with species or class. We tried to compose the host response to Mycoplasma in terms of cellular gene expression. Although gene expression profile of two host HeLa and 293 cells were quite different each other, 30 genes were differentially expressed by mycoplasma infection in both of HeLa and 293 cells. Six of them (PR48, MADH4, MKPX, CRK, RBM7, NEK3) were related to cell cycle or proliferation. Another category of genes like IL1HY1, KLRF1, TNFSF14, GBP1 were host defense to elicit immune responses. With this set of genes, we establish the prediction model for mycoplasma contamination.
Cell Cycle ; Cells, Cultured* ; Gene Expression ; Mycoplasma Infections* ; Mycoplasma* ; Oligonucleotide Array Sequence Analysis ; Transcriptome

The author observed clinical study and response of -interferon treatment of 43 cases of chronic hepatitis who were admitted to Department of Pediatrics Pusan National University College of Medicine during the period of 4 year 6 months from Jenuary, 1988 to Jun, 1992. The results were as follows: 1) Of 43 patients with chronic hepatitis, CPH was 17 cases (39.5%) and CAH was 26 cases (60.5%). 2) Of 43 patients, chronic hepatitis caused by HBV was 37 cases (86.0%) 2 cases were by Wilson's disease, 1 case was by glycogen storage disease and in the 3 cases, the cause was undetermined and CPH by HBV and CAH were 14 cases (82.4%), 23 cases (88.5%) respectively. 3) A peak frequency (46.5%) of chronic hepatitis occurred between 6 to age and male predominated in a propotion of 3.5:1. 4) On the clinical manifestation, jaundice and hepatomegaly was higher frequency on CAH than on CPH. 5) On the laboratory findings, CAH had higher serum ALT and direct bilirubin level than CPH. 6) On effect of -interferon treatment for patients of CAH, serum ALT and AST levels normalized in 81.8% of patients, negative conversion of HBeAg was 38.5%, Anti-HBe serocoversion was 7.7%, negative conversion of HBsAg was 7.7% and the clearnce of HBV-DNA from seum showed 60.0%. 7) The HBeAg seroconversion (28.6%) on children of HBsAg(+) mother showed lower reponse than HBeAg seroconversion (50.0%) of HBsAg(-) mother after -interferon treatment for patients of CAH.
Bilirubin ; Busan ; Child* ; Glycogen Storage Disease ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens ; Hepatitis B, Chronic* ; Hepatitis, Chronic* ; Hepatolenticular Degeneration ; Hepatomegaly ; Humans ; Jaundice ; Male ; Mothers ; Pediatrics

Microarray technology makes it possible to measure the expressions of tens of thousands of genes simultaneously under various experimental conditions. Identifying differentially expressed genes in each single experimental condition is one of the most common first steps in microarray gene expression data analysis. Reasonable choices of thresholds for determining differentially expressed genes are used for the next-step-analysis with suitable statistical significances. We present a supervised model for identifying DEGs using pathway information based on the global connectivity structure. Pathway information can be regarded as a collection of biological knowledge, thus we are trying to determine the optimal threshold so that the consequential connectivity structure can be the most compatible with the existing pathway information. The significant feature of our model is that it uses established knowledge as a reference to determine the direction of analyzing microarray dataset. In the most of previous work, only intrinsic information in the miroarray is used for the identifying DEGs. We hope that our proposed method could contribute to construct biologically meaningful structure from microarray datasets.
Dataset* ; DNA* ; Gene Expression* ; Hope ; Oligonucleotide Array Sequence Analysis* ; Statistics as Topic ; Transcutaneous Electric Nerve Stimulation

Objective: To observe the role of rmh-TNF in enhancing the anti-angiogenesis effect of cisplatin on Lewis lung carcinoma in the mice.Methods: Lewis lung carcinoma model was established in C57BL/6 mice.Sixty model mice were randomly divided into 4 groups: control group,rmh-TNF group(1500000 U/kg),cisplatin group(6.15 mg/kg), and rmh-TNF plus cisplatin group.Twelve days after implantation of cancer cells,different drugs were injected intra- tumorallv for 3d.The expression of hypoxia inducible factor-1?(HIF-1?)gene in the tumor was identified by RT-PCR. Immunohistochemistry(IHC)image analysis was performed to determine the vascular endothelial growth factor(VEGF) and kinase domain region receptor(KDR)expression and the microvessel density(MVD).Expression of matrix metallo- proteinase-2(MMP-2)was detected by flow cytometry.Results: The MVD values in the control group,the rmh-TNF group,the DDP group and the combination group were(24.76?1.28),(18.95?1.22),(19.53?1.15),(10.43?1.05),respectively,with those of the rmh-TNF and DDP groups significantly lower than that of the control group and higher than that of the combination group(all P

Macrophage-associated inflammation is crucial for the pathogenesis of diverse diseases including metabolic disorders. Rhodanthpyrone (Rho) is an active component of Gentiana rhodantha, which has been used in traditional Chinese medicine to treat inflammation. Although synthesis procedures of RhoA and RhoB were reported, the biological effects of the specific compounds have never been explored. In this study, the anti-inflammatory activity and mechanisms of action of RhoA and RhoB were studied in lipopolysaccharide (LPS)-stimulated macrophages. Pretreatment with RhoA and RhoB decreased inducible nitric oxide synthase and cyclooxygenase-2 expressions in RAW 264.7 cells and in thioglycollate-elicited mouse peritoneal macrophages. In addition, it downregulated transcript levels of several inflammatory genes in LPS-stimulated RAW 264.7 cells, including inflammatory cytokines/chemokines (Tnfa, Il6, and Ccl2) and inflammatory mediators (Nos2 and Ptgs2). Macrophage chemotaxis was also inhibited by treatment with the compounds. Mechanistic studies revealed that RhoA and RhoB suppressed the nuclear factor (NF)-κB pathway, but not the canonical mitogen activated protein kinase pathway, in LPS-stimulated condition. Moreover, the inhibitory effect of RhoA and RhoB on inflammatory gene expressions was attenuated by treatment with an NF-κB inhibitor. Our findings suggest that RhoA and RhoB play an anti-inflammatory role at least in part by suppressing the NF-κB pathway during macrophage-mediated inflammation.
Animals ; Chemotaxis ; Cyclooxygenase 2 ; Gene Expression ; Gentiana ; Inflammation ; Interleukin-6 ; Macrophages ; Macrophages, Peritoneal ; Medicine, Chinese Traditional ; Mice ; Nitric Oxide Synthase Type II ; Protein Kinases ; RAW 264.7 Cells

PURPOSE: Transfusion-transmitted virus(TTV) is an newly described nonenveloped human virus, with a circular, negative stranded DNA genome. Although a high prevalence of TTV infection in the normal population has been demonstrated, there is a still possibility of association with hepatitis according to the genotype of TTV. The aim of this study is to investigate the prevalence of TTV infection in Korean children. METHODS: Nested polymerase chain reaction(PCR) using priner sets generated from the noncoding region(NCR) of the viral genome was done in 105 children without liver disease, aged 0-15 years. We performed a second set of PCR using N22 primer in 88 children after the first set of PCR. RESULTS: The TTV DNA was detectable in 36(34%) of 105 children without hepatitis by 5'NCR primer. The prevalence of TTV varied with age:<1 y,16%(4/25); 1-3 y, 44%(15/31); 4-6 y, 31%(5/ 16); 7-9 y, 25%(3/12); 10-15 y, 14%(3/21). By using N22 primers, the prevalence of TTV DNA in children without hepatitis was 11.3%(11/88):<1 y 8%(2/25); 1-3 y, 13.7%(4/29); 4-6 y, 6.2%(1/16); 7-9 y, 33.3%(2/6); 10-14 y, 8.2%(1/12). CONCLUSION: Our result showed a high prevalence of TTV infection, varying with age, in Korean children. Further evaluation of genotypes of TTV in patients with hepatitis and normal children is needed.
Child* ; DNA ; Genome ; Genome, Viral ; Genotype ; Hepatitis ; Humans ; Liver Diseases ; Polymerase Chain Reaction ; Prevalence* ; Torque teno virus*

The analysis of DNA microarry data is one of the most important things for functional genomics research. The matrix representation of microarray data and its successive 'optimal' incisional hyperplanes is a useful platform for developing optimization algorithms to determine the optimal partitioning of pairwise proximity matrix representing completely connected and weighted graph. We developed Deterministic Annealing (DA) approach to determine the successive optimal binary partitioning. DA algorithm demonstrated good performance with the ability to find the 'globally optimal' binary partitions. In addition, the objects that have not been clustered at small non-zero temperature, are considered to be very sensitive to even small randomness, and can be used to estimate the reliability of the clustering.
Cluster Analysis ; DNA* ; Gene Expression* ; Genomics ; Oligonucleotide Array Sequence Analysis*

PURPOSE: The subconjunctival sparganosis is rare tissue helminthiasis which developed frequently in abdominal, urethral, ureteral and vertebral cases. Authors report a case of sparganosis in the subconjunctival mass. METHODS: A 51-year-old man with reddish non painful mass in his left eye on the subconjunctiva that was unresponsive to medications for 6months was evaluated and treated surgically. On surgical exploration, a white thread-like plerocercoid 0.2 x 4.7 cm in size was extracted from the subconjunctival mass and actively motile in the bottle filled with saline. It was confirmed as a typical sparganum under histologic examination. Three months after surgery, the lesion disappeared completely. RESULTS: Although rare, parasitic disease should be suspected in a palpable subconjuntival mass unresponsive to the medical treatment.
Helminthiasis ; Humans ; Middle Aged ; Parasitic Diseases ; Sparganosis* ; Sparganum ; Ureter

BACKGROUND: After peripheral nerve injury in human, a syndrome of events (spontaneous pain, allodynia and hyperalgesia) may be observed that includes no response of morphine and dependency of this pain state on intact sympathetic function. Spinally delivered 2-adrenoceptor agonist and cholinergic agonist or cholinesterase inhibitors have been shown to have actions attenuating the hyperalgesia in rat models of nerve injury-induced pain. Using a fixed-dose analysis and an isobolographic paradigm, the spinal interaction between the 2-adrenoreceptor agonist, clonidine and cholinergic agonist, carbachol is characterized in rat model of nerve injury-induced tactile hyperalgesia. METHODS: Male Sprague Dawely rats were anesthetized with halothane, and the left L5 and L6 spinal nerve were ligated (Chung model). After recovery, a polyethylene tubing catheter was implanted into lumbar intrathecal space. After recovery from catheter implantation, intrathecal dose-response curves were established for the antiallodynic effect of carbachol (0.1, 0.3, 1.0, 3.0 microgram) and clonidine (0.3, 1.0, 3.0, 10 microgram) alone to obtain the ED50 for each agent. ED50 fractions (1/2, 1/4, 1/8, 1/16) of drug combinations of carbachol-clonidine were administered and thresholds for left hind limb paw withdrawal to von Frey hair application were assessed. The ED50 of carbachol-clonidine combination was established and isobolographic analysis of the drug interactions was carried out.c RESULTS: Intrathecal carbachol and clonidine alone produced dose-dependent reductions of tactile allodynia: ED50 of 66 ng (12~367 ng) and 39 ng (1~1452 ng), respectively. With the fixed dose analysis, the log dose-response curves showed a left shift that considerably exceeds the theoretical curves made by a simple sum of the effects of carbachol alone and clonidine. With the isobolographic analysis, ED50 of mixture was found to be statistically less than the theoretical additive ED50 of mixture. CONCLUSION: The experiments suggest that intrathecal carbachol and clonidine alone produce a dose dependent antagonism on touch evoked allodynia and intrathecal carbachol is synergistic when combined with intrathecal clonidine.
Animals ; Carbachol* ; Catheters ; Cholinergic Agonists ; Cholinesterase Inhibitors ; Clonidine* ; Drug Combinations ; Drug Interactions* ; Extremities ; Hair ; Halothane ; Humans ; Hyperalgesia ; Male ; Models, Animal ; Morphine ; Neuralgia* ; Peripheral Nerve Injuries ; Polyethylene ; Rats* ; Spinal Nerves