12 strains of H2-producing bacteria were isolated and purified from anaerobic sludge, aerobic sludge and river bottom sludge by Hungate method. A new species of high efficient hydrogen production bacterium Enterococcus sp. LG1 (registration number: EU258743 ) was studied deeply. It was showed that the Enterococcus sp. LG1 was an anaerobic and Gram-negative bacterium. Sequence analysis of this type of clones showed that it was affiliated with the genus Enterococcus and it was not reported yet in other paper at present. Meanwhile, batch tests of anaerobic fermentative hydrogen production by Enterococcus sp. LG1 were investigated by using sterilization pretreated sludge as substrate. The changes of soluble COD, protein, carbohydrate and pH value during hydrogen fermentation were monitored. It was found that only hydrogen and carbon dioxide were produced by this strain and no methane was detected during fermentation. The maximal hydrogen yield was 36.48 mL/g TCOD and the hydrogen concentration in the gas phase was 73.5%. The Enterococcus sp. LG1 was a butyrate fermentation bacteria analyzed by metabolites.

OBJECTIVETo investigate the efficacy of surgical management for pyloric stenosis induced by gastrointestinal chemical burn in children.

METHODSClinical data of 11 children with pyloric stenosis induced by gastrointestinal chemical burn were analyzed retrospectively. After the failure of medicine, intervention of low balloon expansion and stent placement, they underwent pylorectomy and gastroduodenostomy. The body weight, height, serum albumin, hemoglobin, transferrin were compared between 1 day before and 3 months after operation.

RESULTSThere were 10 males and 1 female with a mean age of 4.5 years old. The main cause of serious pyloric stenosis was the wrong intake of hydrochloric acid. Lesions involved the esophagus and stomach in the early stage, and 4 weeks later, the lesion mainly involved the pylorus, which resulted in scarring pyloric stenosis and complete pyloric obstruction. Pylorectomy and gastroduodenostomy was successfully performed. The mean operative time was (134±26) min. The estimated blood loss was (5±2) ml. The postoperative length of stay was (10±3) d. There were no surgical complications. During the follow-up of 3 months, all the patients resumed regular diet. The height, body weight, and intelligence appeared to be normal. They showed significant improvement in weight, serum albumin, globulin, hemoglobin, transferrin at 3 months after the surgery(P<0.05). Six months after surgery, the anastomosis was shown to be nornal in barium follow through exam with no signs of stricture of ulcer.

CONCLUSIONPylorectomy and gastroduodenostomy is an effective management for pyloric stenosis induced by gastrointestinal chemical burn in children, whose short-term efficacy is good.

Burns, Chemical ; Child ; Gastrectomy ; Gastroenterostomy ; Humans ; Pyloric Stenosis ; Pylorus ; surgery

OBJECTIVETo observe the effects of siwu tang on protein expression of bone marrow of blood deficiency mice and provide the theoretical and experimental basis for understanding the molecular mechanism of blood enriching function of siwu tang.

METHODBlood deficiency mice were established by using 3.5 Gy 60Co gamma-ray. With proteomic technologies including two-dimensional electrophoresis, image analysis, in-gel digestion, peptide mass fingerprinting and bioinformatics the proteins of bone marrow of blood deficiency mice were isolated, analyzed, and identified.

RESULTSiwu tang could reverse 10 up-regulated and 4 down-regulated proteins of blood deficiency mice bone marrow. Seven of the proteins were likely to be lymphocyte specific protein 1, proteasome 26S ATPase subunit 4, hematopoietic cell protein-tyrosine phosphatase, glyceraldehyde-3-phosphate dehydrogenase, growth factor receptor binding protein 14 and lgals12 respectively.

CONCLUSIONSiwu tang can regulate the protein expression of bone marrow of blood deficiency mice and thus promote the growth and differentiation of hematopoietic cells and then exert its effects on blood enriching function.

Adaptor Proteins, Signal Transducing ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Phosphoproteins ; blood ; Proteins ; metabolism ; Proteome ; metabolism ; Proteomics ; methods ; Radiation Injuries, Experimental ; blood ; Whole-Body Irradiation

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.
Cell Survival ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; Cytochrome P-450 CYP1A1 ; genetics ; Diosgenin ; analogs & derivatives ; toxicity ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; secretion ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Receptors, Aryl Hydrocarbon ; genetics

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

OBJECTIVETo investigate the distribution patterns and proliferative activity of lymphatic vessels in colorectal carcinomas (CRC) and their relationship with tumor metastasis and disease prognosis.

METHODSThe microlymphatic density (MLD) and microvascular density in tumoral and non-tumoral areas of 96 cases of CRC were evaluated by immunohistochemistry, using monoclonal antibodies for podoplanin and CD34 respectively. The Ki-67 expression of the lymphatic and blood vessels was detected by double-labeling immunohistochemistry. The relationship between MLD and clinicopathologic features and prognosis was analyzed.

RESULTSThe lymph vessels at central and superficia1 portions of CRC often had a reticular architecture with numerous tiny and ill-defined lumina, while those at the tumor borders had large and open lumina. The MLD at tumor borders (51.2 +/- 25.5) was significantly higher than that in normal colorectal mucosa (29.4 +/- 9.0) and other portions of CRC (P < 0.01). The Ki-67 labeling index of the lymphatic lining cells at tumor borders (0.23 +/- 0.17) was significantly higher than that in other portions of CRC (P < 0.05). The MLD significantly correlated with lymphatic involvement by tumor cells, regional lymph node metastasis and distant metastasis (P < 0.01). The 5-year survival rate was also significantly lower in patients with high MLD (P < 0.05).

CONCLUSIONSNeolymphatic vessels are commonly seen in CRC, especially at tumor borders. High MLD at tumor borders is associated with metastasis. The detection of MLD at tumor borders may thus be useful in predicting lymph node metastasis and prognosis in patients with CPC.

Adenocarcinoma ; immunology ; pathology ; physiopathology ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; immunology ; pathology ; physiopathology ; Endothelium, Vascular ; immunology ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Rate

OBJECTIVE: To investigate the biological characteristics of endothelial progenitor cells (EPCs) from the umbilical cord blood (UCB), and to evaluate their oncogenicity after long-term culture in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the UCB and cultured in MCDB131 medium supplemented with 20% FBS, VEGF and other growth factors. Morphology of the EPCs was observed, and the growth curve of the EPCs was investigated. Surface antigens of the EPCs were analyzed by the flow-cytometer. The capability of intaking the acetylated low-density lipoprotein (acLDL) of the EPCs was detected using fluoresencent chemical method. The vasoformative capability and genetic stability of EPCs were cultured in matrigel, and examined by karyotype analysis. The oncogenicity of EPCs was verified by the tumorigenesis test in athymic mouse and soft agar. RESULTS: EPCs were successfully derived from the UCB, and could be passaged to at least 42(nd) generation and had strong abilities of proliferation, acLDL intake and vasoformation, but there was not oncogenicity. They expressed endothelial cell-surface antigens and maintained normal karyotype. CONCLUSION The EPCs with proliferative potential can be isolated from the UCB. They can be passaged in long-term cultures without oncogenicity, and can maintain normal karyotype. The EPCs can be served as a new type of cells in cell and gene therapy.
Animals ; Antigens, Surface ; analysis ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; HeLa Cells ; Humans ; Infant, Newborn ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Karyotyping ; Mice ; Mice, Nude ; Neoplasms, Experimental ; pathology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo make preliminary study on the chemical foundation of hematopoietic effect of si-wu-tang.

METHODSi-wu-tang was fractionated by using precipitation method with ethanol and liquid-liquid extraction. Activity was investigated on radiated mice as model of blood deficiency.

RESULTAt first si-wu-tang was separated into three fractions, fraction A, B and C. Activity study showed that fraction B and C could raise the amount of peripheral white blood cell in radiated mice. Then fraction B and C were respectively further separated into fraction B1, B2, B3 and fraction C1, C2, C3. Activity study showed that fraction C2, C3 and B3 could raise the amount of peripheral white blood cell in radiated mice. Furthermore, effect of fraction C1, C2, C3, B and paeoniflorin on hematopoietic progenitor cell in bone marrow of radiated mice were investigated. Fraction C2, C3, B and paeoniflorin could promote the proliferation of hematopoietic progenitor cell in bone marrow of radiated mice.

CONCLUSIONFraction C2, C3, B and paeoniflorin are responsible for hematopoietic activity of si-wu-tang.

Angelica sinensis ; chemistry ; Animals ; Benzoates ; isolation & purification ; pharmacology ; Bone Marrow Cells ; radiation effects ; Bridged-Ring Compounds ; isolation & purification ; pharmacology ; Cell Proliferation ; drug effects ; radiation effects ; Colony-Forming Units Assay ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Glucosides ; isolation & purification ; pharmacology ; Hematopoietic Stem Cells ; cytology ; Leukocyte Count ; Ligusticum ; chemistry ; Mice ; Mice, Inbred C57BL ; Monoterpenes ; Paeonia ; chemistry ; Rehmannia ; chemistry ; Whole-Body Irradiation

OBJECTIVETo study the role of O(6)-methylguanine-DNA methyltransferase (hMGMT) in the development of human lung cancer.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) method was applied to measure hMGMT mRNA expression in 150 lung cancer specimens, 40 normal lung tissues, and in the peripheral mononuclear blood cells from 50 lung cancer cases and 50 normal controls. The protein expressions of p53, C-MYC and K-RAS were assessed by immuno-histochemistry. The effects of some exposure factors on the expression of hMGMT gene were analyzed. The relationships between hMGMT gene and cancer related genes p53, C-MYC and K-RAS were investigated.

RESULTSThe mRNA of hMGMT was low or absent in 49 of 150 (32.7%) lung cancer specimens, whereas 2 of 40 (5%) normal lung tissues had reduced the levels of hMGMT mRNA. The low expression of hMGMT seemed to be a risk factor of lung cancer, with a OR of 9.22 (2.05-57.65). Reduced expression levels of hMGMT mRNA were observed in 10 of 50 (20%) lung cancer patients' peripheral mononuclear blood cells, and 2 of 50 (4%) blood cells among normal controls. When investigating the exposure factors which affecting the expression of hMGMT gene, we noticed that smoking was suppressing the expression of hMGMT gene. Interestingly, over-expression of K-RAS oncogene was significantly correlated with low expression of hMGMT (P < 0.05). However, the expressions of p53 and C-myc were not correlated with the status of hMGMT gene.

CONCLUSIONhMGMT might play an important role in the development of human lung cancer. Low expression of hMGMT gene seemed to be a risk factor for lung cancer which could be used as a valuable biomarker on susceptibility of human lung cancers.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; enzymology ; genetics ; China ; epidemiology ; DNA Repair ; genetics ; Female ; Genes, ras ; genetics ; Humans ; Lung Neoplasms ; enzymology ; genetics ; Male ; Middle Aged ; O(6)-Methylguanine-DNA Methyltransferase ; biosynthesis ; genetics ; metabolism ; Point Mutation ; RNA, Messenger ; biosynthesis ; genetics ; Smoking ; adverse effects ; ras Proteins ; biosynthesis ; genetics