Objective To search for the reasons of high positive rate of amotile bacteria and the diagnosis of septicemia in new-born Methods The blood was drawn from the different site of the new-born with septicemia and carricd out blood culture. The drug sensitivity test had been done by the method of paper stripdiffusion. The plasmids of bacteria were extracted rapidly by medified Birnboim method and the plasmid analyss was carried out. The plasmids's DNA of 35 epidemic strain was cut off by both restriction enzyme of Hind Ⅲ and EcoR Ⅰ. The outer membrane protein (OMP) was determined by SDS-polyacrylamide gel electrophoresis.Results There are 51 patients with positive blood culture amotile bacterium,of them, pollution; 35 cases (68．6%), septicemia: only 16 cases (31．4%),54．8% (57/104) strains bacteria have drug resistance to more of 12 drugs. 87．3% (165/189) strains bacteria have plasmids. They are cut off as 6 DNA fragments (1．9,2,4,5, 8．5 and 18Kb) by Hind Ⅲ restrietion enzyme. and as 5 DNA fragments (2,2．6,3．2, 6．3 and 22 Kb) by EcoR Ⅰrestrietion enzyme, it is showed that they come from a same clone. The epidemic strain include 10 slips OMP, but non-epidemic strain have 11 slip OMP, increase a 25Kd belt. The amotile bacteria with above-mentioned plasmid spectrum, restriction enzyme spectrum and OMP spectrum are only seen in the air, therapeutic dish and syringe needle.Conclusion The pollution is an important reason of amotile bactorium high positiye rate in new-born.Diagnosing septicemia should depend on bacteria culture, plasmid analysis restriction enzyme analysis of plasmid DNA, oMP determination and combining medical history and clinical manifestation.

Acupuncture Therapy ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pterygium ; therapy ; Yin Deficiency ; therapy

OBJECTIVE Vascular dementia (VD) refers to a progressive decline in memory and cognitive function caused by chronic cerebral ischemia. 2-Vessels occlusion (2-VO) has been widely used as a model of VD. Xiao-Xu-Ming decoction, a well-known traditional Chinese medicine prescrip-tion,has been widely used to treat stroke and sequelae of stroke.The present study was to investigate the mechanism of Xiao-Xu-Ming decoction(XXM) against chronic cerebral ischemia injury in rats. METHODS After XXM treatment, rats were performed a memory testing with Morris water maze and motor ability testing using prehensile test and inclined screen test.Neuronal plasticity was observed by immunofluorescent staining with MAP2 antibody. Differentially expressed proteins of rat hippocampus were analyzed by Label-free quantitative proteomics. RESULTS XXM significantly alleviated 2-VO-induced learning and memory deficits, motor ability dysfunction, and neuronal plasticity injury in rats. The mechanism might be involved in up-regulation of 39 proteins and down-regulation of 13 proteins in the hippocampus of rats after XXM treatment vs 2-VO group rats.Gene ontology and pathway analysis showed that the regulated proteins are mainly involved in oxidation reduction process, intracellular signaling cascade process, and protein catabolic process, etc. The signal pathways are mainly involved in ubiquitin mediated proteolysis and phosphatidylinositol signaling system. CONCLUSION Current findings provide new insights into the molecular mechanisms of XXM on chronic cerebral ischemia.

BACKGROUNDThe antitumor role of Ras association domain family 1A (RASSF1A) gene and its potential molecular mechanisms are not well understood. The objective of this study was to observe the antitumor ability of RASSF1A in hepatocellular carcinoma, and study the mechanisms of cell apoptosis induced by RASSF1A.

METHODSAfter stably transfecting a RASSF1A (wild-type or mutant) expression vector into the BEL-7402 hepatocellular carcinoma cell line, RT-PCR and Western blotting was used to detect the RASSF1A expression levels in recombinant cells. The effects of wild-type RASSF1A on cell growth were observed in vitro by analyzing cell proliferation rate, cell colony formation, and in vivo by analyzing tumorigenesis in nude mice. In addition, the effect of RASSF1A gene expression on the chemosensitivity of human hepatocellular carcinoma cells to antitumor drugs was examined by inhibition of cell proliferation and the percentage of apoptotic cells.

RESULTSWild-type RASSF1A, not the mutant, suppressed cell growth in vitro and in vivo. Re-expression of wild-type RASSF1A could enhance the inhibition of cell proliferation and the percentage of apoptotic cells following cell treatment with mitomycin, but had no significant effect when combined with adriamycin, etoposide, 5-fluorouracil and cisplatin treatment.

CONCLUSIONWild-type RASSF1A inhibits cell growth and enhances cell chemosensitivity to mitomycin in hepatocellular carcinoma, suggesting that RASSF1A may serve as a new target for gene therapy in hepatocellular carcinoma patients.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Blotting, Western ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Genetic Therapy ; methods ; Humans ; Mitomycin ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; genetics ; metabolism ; physiology

OBJECTIVETo screen the severe acute respiratory syndromes (SARS) mimotopes with random phage peptide library and to investigate their immunogenicity.

METHODSUsing SARS sera as selective molecule, a 12 mer phage peptide library was biopanned and positive clones containing the mimic epitopes were selected. The immuno-characteriation of the epitopes were then investigated.

RESULTS2 positive clones that having specific affinity to SARS sera were obtained. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the sequence of SARS.

CONCLUSIONSARS mimotopes were obtained by phage peptide library screening. This method might provide a new approach for SARS therapy and vaccine development.

Animals ; Antigens, Viral ; Base Sequence ; Biomimetic Materials ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; genetics ; immunology ; Humans ; Peptide Library ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; immunology

This study was aimed to explore the correlation of mean platelet volume (MPV), fibrinogen (FIB) and blood rheology with the youth patients with cerebral infarction, so as to provide the basis for the clinical early diagnosis and treatment. The 109 patients with cerebral infarction aged between 18 - 45 were divided into three group: the large (> 10 cm(3)), middle (4 - 10 cm(3)) and small (< 4 cm(3)) area infarction; 30 healthy persons were served as control group. All the four groups were subjected to 16 examinations, such as MPV, FIB, and rheology (Lηb, Mηb, Hηb, ηp, Lηr, Mηr, Hηr, KVE, EAI, ERI, EDI, EEI, HCT, ESR). The results showed that all the MPV, FIB and rheology indexes of the different infarction groups were higher than those of healthy control group (P < 0.05). The MPV, FIB and rheology indexes in the large area infarction group were all higher than those in the small area infarction group (P < 0.05). The indexes of MPV, FIB and rheology in the various cerebral infarction area groups obviously decreased, but those did not reach to the level in the healthy control group (P < 0.05). The MPV, FIB content and rheology level correlated with infarction areas (r = 0.36, 0.29 and 0.48, respectively). It is concluded that the serious intensity of youth patients with cerebral infarction positively correlated with the levels of MPV, FIB and rheology indexes. Regular examination of above mentioned index may be useful to prevent youth patients from cerebral infraction.
Adolescent ; Adult ; Blood Platelets ; Case-Control Studies ; Cerebral Infarction ; blood ; diagnosis ; Female ; Fibrinogen ; metabolism ; Hemorheology ; Humans ; Male ; Middle Aged ; Platelet Count ; Young Adult

OBJECTIVETo study the alteration of protein Z (PZ) in patients with cardio-cerebral thrombotic diseases, its clinical significance and relations with FX.

METHODSPZ and FX:Ag were measured by ELISA, and plasma FX:C by first stage method. In 170 patients with acute ischemic stroke (AIS), 40 acute myocardial infarction (AMI) and 60 healthy adults as contrast, PZ, FX:C and FX:Ag were measured and compared between incipience and recurrence, different ages and genders.

RESULTSIn AIS and AMI groups, PZ levels decreased significantly to (940.02 +/- 229.82) microg/L and (1071.44 +/- 180.52) microg/L, respectively \[the contrast group was (2257.97 +/- 479.76) microg/L, P < 0.001\]. But FX:C and FX:Ag raised to (136.73 +/- 34.93)% and (135.54 +/- 54.39)% in AIS group; and to (139.53 +/- 29.18)%, (129.75 +/- 21.91)% in AMI group, respectively, while in the contrast group they were (94.33 +/- 22.00)% and (77.22 +/- 13.19)% (P < 0.001). In the comparative research between the AIS group, AMI group and the contrast group, PZ level was clearly found to negatively relate to the level of FX:C and FX:Ag (P < 0.001). Meanwhile, PZ level, FX:C and FX:Ag in recur-AIS group and recur-AMI group exhibited significant differences (P < 0.05) from those in the primary AIS and AMI groups, suggesting that the decrease of PZ levels reflected the pathological process of the disease. In addition, PZ level gradually decreased with the increase of age (P < 0.05), while FX:C and FX:Ag had no relations with age (P > 0.05). No correlation was found in sex with PZ level, FX:C, FX:Ag (P > 0.05).

CONCLUSIONPZ level was significantly decreased in AIS and AMI patients and was negatively related to FX:C and FX:Ag. The mechanism leading to FX increase may partially related with the decreased of PZ. PZ level was different in the primary and recurrent disease and was gradually decreased with the increase of age. Lack of PZ might be a etiological factor of cardio-cerebral thrombotic diseases.

Aged ; Aged, 80 and over ; Blood Proteins ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Factor X ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; Stroke ; blood

OBJECTIVETo analyze the molecular characters of the W135 Neisseria meningitidis strain firstly isolated from patients in Guangdong province.

METHODSBiochemical profile by using the API NH system (bio-Merieux, France) was used for confirmation,and sero-grouping of the meningococcal isolates including one serogroup W135, one serogroup C and three serogroups of a Neisseria meningitidis isolated from patients in Guangdong province in recent two years were performed. The subtype was determined after amplifying porA and porB respectively from the genome DNA of Neisseria meningitidis. Multilocus sequence typing (MLST) was performed for determining the allele profiles and the sequence types (STs). The polygenetic tree was obtained by analyzing the allele profiles with program Splits tree online. The molecular characters of the serogroup W135 Neisseria meningitidis was analyzed by its evolution relationship and the variable regions in porA and porB which encoding the outer membranes proteins (OMPs).

RESULTSThe subtype determined by porA variable regions of the serogroup W135 Neisseria meningitidis was P1.5,2, which was one of the most invasive types. The types of variable regions (VRs) I, IV, V, VII with porB were 1, 1, 1, 17, and there was no VI and VIII in porB. The allele profile of the W135 strain in this study was 2, 123, 4, 3, 8, 4, 6, and its sequence type was ST-2960, which belonged to ST-11/ET-37 clone complex. The subtypes of the serogroup C and serogroup A strains were P1.20, while their types of VR IV were all 7, and they all hadn't VR VII in porB. The strain serogroup C belonged to ST-4821 clone complex, and the 3 serogroup A strains belonged to ST-5 clone complex.

CONCLUSIONThe molecular character of the serogroup W135 Neisseria meningitidis should be the same with the strains isolated in foreign country, and be different from the epidemic types in the area. This serogroup W135 Neisseria meningitis isolated from patients in Guangdong for the first time was thought to be a new type appearing in the local area.

Bacterial Typing Techniques ; China ; epidemiology ; DNA, Bacterial ; Disease Outbreaks ; Encephalomyelitis ; epidemiology ; microbiology ; Genotype ; Humans ; Molecular Sequence Data ; Neisseria meningitidis, Serogroup W-135 ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA

OBJECTIVETo elucidate intracellular signal pathway in formation of multidrug resistance (MDR) of hepatocellular carcinoma (HCC) induced by its microenvironment, and to explore the potential role of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway in this process.

METHODSActivity of ERK/MAPK was examined by Western blot technique through comparing the ratio of phosphorylation of ERK/MAPK to total ERK/MAPK protein in HepG2 cells exposed to hypoxia, low glucose or transfected by plasmid pcDNA3/HBX. After being treated by the specific ERK/MAPK pathway inhibitor U0126, Western blot technique was used to analyze the alterations of the expression of P-gp, MRP1, LRP and HIF-1alpha at protein level. RT-PCR was used to analyze the alterations of the expression of HIF-1alpha mRNA. Cellular location of HIF-1alpha protein was determined by immunocytochemistry after being treated by U0126.

RESULTSThe activations of ERK/MAPK determined by the ratio of phosphorylated ERK/MAPK to the total ERK/MAPK were increased in varying degrees in HepG2 cells respectively exposed to different microenvironment. After being treated by U0126 for 12 h, the expressions of mdr1, MRP1, LRP genes and protein in those cells were decreased to some extent. However, the gene expression of HIF-1alpha was not influenced and only its protein was decreased. HIF-1alpha protein was reversely translocated into cytoplasm from nucleus after being treated by U0126.

CONCLUSIONSERK/MAPK pathway is involved in the course of the formation of MDR of HCC induced by microenvironment.

Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MAP Kinase Signaling System ; physiology ; Mitogen-Activated Protein Kinases ; metabolism ; Multidrug Resistance-Associated Proteins ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the reversal of multidrug resistance in the cell line HepG2/ADM induced by TNF-alpha.

METHODSHepG2/ADM cells were incubated with different concentrations of TNF-alpha (100, 500 and 2500 U/ml) for 72 h. Real-time PCR was performed to compare the mRNA levels of MDR1 with PPAR-alpha in the different concentrations of TNF-alpha treated cells. The Annexin V assay was used to check cell apoptosis induced by 0.5 mg/L adriamycin. Rhodamine 123 efflux assay and MTT assay were used to study P-gp activity and drug resistance in each group, respectively.

RESULTSTNF-alpha could induce down-regulation of MDR1 and up-regulation of PPAR-alpha. Meanwhile, it could enhance cell cytotoxicity and cell apoptosis induced by 0.5 mg/L adriamycin.

CONCLUSIONSTNF-alpha could partially reverse the multidrug resistance of HepG2/ADM cells by down-regulating the expression of MDR1 and up-regulating the expression of PPAR-alpha.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; PPAR alpha ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology