To study the possible mechanism of the effect of glucagon-like peptide-1 (GLP-1) on injury to neonatal rat cardiomyocytes induced by hypoxia-reoxygenation. Lactate dehydrogenase activity [(210.0±11.5) vs (101.4±6.5) U/L] ,apoptosis rate [(8. 138±1. 512) vs(0. 575±0. 168)%] ,and caspase-3 activity [(44.52± 5.69)vs(19.98±1.97) ,all P<0.01] were all increased after hypoxia-reoxygenation. GLP-1 appears to directly act on cardiomyocytes and to protect them from hypoxia-reoxygenation injury [lactate dehydrogenase (190.2±9.0) U/ L, apoptosis rate (2.688±0.580) %, caspase-3 activity 30.34±4.18] mainly by inhibiting the apoptosis probably via the PBK/Akt signaling pathways.

Objective:To investigate the effect of Rab7 on cytokine induced by TLR7 (Toll like receptor-7) R848 activated in Raw264.7,and discusses the influence of Rab7 on MAPK signal transduction.Methods: TLR7 downstream cytokines such as TNF-α,IL-6,IFN-α,IFN-β and IP-10 activated by R848 were detected through Q-PCR in Rab7 silenced mouse macrophages,and then analysis of phosphorylation of MAPK determined with Western blot showed the effect of Rab7 on signal transduction of MAPK.Results: Rab7 inhibit production of cytokine activated by TLR7,and also,Rab7 had an inhibitory effect on MAPK signal pathway.Conclusion: The experimental results further illustrate that the Rab7 is the TLR7 signal transduction pathway negative regulatory factor,and to participate in MAPK signaling pathway.

BACKGROUNDOur previous study had cloned two glioma cell lines SWOZ1 and SWOZ2 isolated from parental glioma cell line SWO38. The 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) resistance of SWOZ1 was higher than that of SWOZ2. Since O6-methylguanine-DNA methyltransferase (MGMT) was thought to be closely related to BCNU resistance in glioma, this study aimed to explore the function of MGMT in glioma resistant to BCNU.

METHODSA BCNU resistant glioma cell line SWOZ2-BCNU was established. The expression of MGMT was detected in SWOZ1, SWOZ2 and SWOZ2-BCNU. Small interferencing RNA targeting MGMT was used to silence the expression of MGMT in resistant cell lines SWOZ1 and SWOZ2-BCNU. The cytotoxicity of BCNU to these cells was measured using the cell counting kit-8 assay. Statistical analysis was carried out by one-way analysis of variance in statistical package SPSS 13.0.

RESULTSThe resistance of SWOZ1 and SWOZ2-BCNU against BCNU was 4.9-fold and 5.3-fold higher than that of SWOZ2. The results of quantitative RT-PCR and Western blotting confirmed that MGMT was both significantly increased in SWOZ1 and SWOZ2-BCNU compared to SOWZ2. After transfection with small interferencing RNA targeting MGMT, a decreased level of MGMT mRNA expression in SWOZ1 and SWOZ2-BCNU for more than 75% compared to negative control was found and confirmed by Western blotting. As a result, the resistance against BCNU was reversed for about 50% both in the BCNU-resistant cell lines SWOZ1 and SWOZ2-BCNU.

CONCLUSIONSSilencing MGMT with specific small interferencing RNA can reverse the BCNU resistant phenotype in these glioma cell lines. MGMT may play an important role both in intrinsic and acquired BCNU-resistance in glioma.

Blotting, Western ; Carmustine ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Glioma ; genetics ; metabolism ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; RNA, Small Interfering ; Reverse Transcriptase Polymerase Chain Reaction ; Sincalide ; metabolism

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of Ca2+, Mg2+, K+, and HCO3 - in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.
ATP-Binding Cassette Transporters/*chemistry/*genetics/metabolism ; Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Calcium/metabolism ; *Cloning, Molecular ; Cryptosporidiosis/parasitology ; Cryptosporidium/chemistry/genetics/*metabolism ; Humans ; Iron/metabolism ; Mice ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/*chemistry/*genetics/metabolism ; Sequence Alignment

OBJECTIVETo find out a set of practical,objective, and quantitative laboratory indices of climacteric syndrome (CS) patients of Shen deficiency syndrome (SDS), thus studying the essence of SDS from the perspective of laboratory medicine.

METHODSRecruited were 40 CS patients of SDS (or of SDS as main syndrome) as the SDS group, while another 40 healthy subjects were recruited as the control group. Their serum samples were collected. Serum levels of total triiodothyronine (TT3), total thyroxine (TT4), free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (TESTO), estradiol (E2), prolactin (PRL), progesterone (PROG), cortisol (CORT), immunoglobulin M (IgM), immunoglobulin G (lgG), Complement 3 (C3), complement hemolysis 50% (CH50), angiotensin converting enzyme (ACE), aldosterone (ALD), serum alkaline phosphatase (ALP), and bone Gla-protein (BGP) were measured by automatic electrochemical luminescence assay analyzer, automatic chemiluminescence assay analyzer, automatic biochemistry analyzer, and automatic enzyme-linked immunosorbent assay (ELISA) analyzer. The correlation between syndrome types and laboratory indices were judged by gradual discriminant analyses.

RESULTS(1) Compared with the control group,serum levels of CORT, TESTO, E2, TT3, FT3, FT4, TSH, C3, CH50, ALP, and BGP significantly decreased in the SDS group (P < 0.01, P < 0. 05), while FSH, LH, and ACE significantly increased (P < 0.05). (2) The index with stronger capacity for diagnosing CS patients of SDS was ranked from high to low as CH50, PROG, TSH, TESTO, BGP, CORT, and C3, with their contribution rate of the discriminant function being 95.9%. (3) Discriminant analysis equation of CS patients of SDS was Y = -25.904 - 0.468CH50 + 0.002PROG + 0.182TSH + 9.690TESTO + 1.015BGP + 0.016CORT + 33.581 C3.

CONCLUSIONS(1) CS patients of SDS were closely correlated with thyroid hypothalamus-pituitary-adrenal axis, hypothalamus-pituitary-adrenal axis, renin-renin-angiotensin-aldosterone system,the immune function, and bone formation, and etc. (2) CH50 might be of a high sensibility marker for diagnosing CS patients of SDS. (3) Discriminant analysis equations of laboratory medicine index may be used in preliminary diagnosis and auxiliary certificate of CS patients of SDS.

Case-Control Studies ; Climacteric ; metabolism ; Discriminant Analysis ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Hydrocortisone ; blood ; Luteinizing Hormone ; blood ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Pituitary-Adrenal System ; Progesterone ; metabolism ; Prolactin ; blood ; Renin-Angiotensin System ; Testosterone ; blood ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo study the effects of Buyang Huanwu Decoction ([Chinese characters: see text]) on promoting functional recovery of crushed common peroneal nerve in rats.

METHODSThirty Sprague-Dawley rats were subjected to produce common peroneal nerve injuries model,and the length of injury was 5 mm. All the rats were divided into 3 groups: BYHWD group, mecobalamin group and model group. The drugs were given by gavage daily for 18 days. Footprint test was performed at the 18th day after surgery to evaluate toe spread function (TSF). Electrophysiology was performed at the 18th day after operation to determine the nerve conduct velocity (NCV). The wet weight ratio and section area of tibial muscle were also measured.

RESULTS(TSF:At the 18th day after operation, the TSF in BYHWD group (-0.15 +/- 0.07) increased significantly compared with that of model group (-0.25 +/- 0.07) (P < 0.01); the TSF in mecobalamin group (-0.17 +/- 0.08) also increased notably compared with that of model group (P < 0.01).(2) NCV: the NCV in BYHWD group [(18.36 +/- 2.74) m/s] (P < 0.01l) and in mecobalamin group [(16.32 +/- 3.54) m/s] (P < 0.05) also increased significantly compared with that of model group [(9.08 +/- 2.56) m/s]; there was striking variation between model group and mecobalamin group (P < 0.05). (3) Wet weight ratio: the wet weight ratio in BYHWD group [(64.21 +/- 2.92)%] (P < 0.01)and in mecobalamin group [(62.43 +/- 3.21)%] (P < 0.01) all increased significantly compared with that of model group [(54.27 +/- 2.05)%]. (4) The section area of tibial muscle: the section area of tibial muscle in BYHWD group [(654.21 +/- 42.92) cm2] (P < 0.01) and in mecobalamin group [(638.43 +/- 93.21) cm2] (P < 0.01) all increased significantly compared with that of model group [(574.27 +/- 52.05) cm2]; there was also striking variation between model group and mecobalamin group (P < 0.05).

CONCLUSIONBYHWD can promotes functional recovery of crushed nerve as a result of accelerating recovery of TSF, raising NCV and delaying the decrease of tibial muscle section area and wet weight ratio.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Electrophysiological Phenomena ; drug effects ; Male ; Organ Size ; drug effects ; Peroneal Nerve ; drug effects ; injuries ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; drug effects ; Time Factors

OBJECTIVETo find a convenient and exact method for evaluating acrosome reaction in human spermatozoa.

METHODSThe semen of the normal male was mixed and then divided into 6 groups. Coomassie brilliant blue (CBB) staining, chlortetracycline (CTC) fluorescence staining and acid phosphatase (ACP) detection were used for morphological observation and data analysis of the acrosome status of the human sperm treated with or without progesterone.

RESULTSThere were obvious morphological differences between the acrosome-reaction and acrosome-intact spermatozoa in CBB staining and CTC fluorescence staining, and significant differences were observed between the experimental and control spermatozoa by the three methods (P < 0.05).

CONCLUSIONAll the three methods can be used to assess acrosome reaction in human spermatozoa, but Coomassie brilliant blue (CBB) staining is much more convenient and stable.

Acid Phosphatase ; Acrosome Reaction ; drug effects ; Cells, Cultured ; Chlortetracycline ; Humans ; Male ; Progesterone ; pharmacology ; Rosaniline Dyes ; Spermatozoa ; cytology ; Staining and Labeling ; methods

OBJECTIVETo explore the causative pathogens in littoral hand infections which exhibited chronic granulomatous inflammation, the relationship between chronic granulomatous inflammation and mycobacteria and to discuss the prospects of PCR in clinical application for diagnosis of granulomatous inflammation.

METHODWith 16S-rDNA as the target sequence, Nest-PCR was used to detect mycobacteria directly from 37 cases of chronic granulomatous inflammations, and identified them by gene sequencing.

RESULTSTwenty-four of 37 cases were positive for mycobacteria by Nest-PCR, in which 17 were M.marinum, 1 M.chelonae, 2 M.avium, 2 M.kansasii, and 2 M.tubercular through gene sequencing.

CONCLUSIONSNest-PCR combining gene sequencing proved to be a liable and sensitive method to detect Non-tubercular mycobacteria (NTM) in fresh tissue. NTM is the major factor of hand specific chronic infections other than tubercular. Pathological changes are difficult to differentiate TB from NTM and bacterial evidence was necessary.

Chronic Disease ; DNA, Bacterial ; chemistry ; genetics ; Granuloma ; diagnosis ; microbiology ; Hand ; Humans ; Inflammation ; diagnosis ; microbiology ; Molecular Diagnostic Techniques ; Mycobacterium Infections, Nontuberculous ; diagnosis ; microbiology ; Mycobacterium marinum ; genetics ; isolation & purification ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Nontuberculous Mycobacteria ; genetics ; isolation & purification ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; Sequence Analysis, DNA

OBJECTIVETo study the effect of irradiation on the susceptibility of radiation caries.

METHODSThe structures of 56 teeth enamel and dentin of 63 roots were observed using SEM and the collagen fibre and the resistance to the acid were also investigated after irradiation of 30 Gy, 50 Gy and 70 Gy.

RESULTSThe enamel structure changes were found after irradiation with different doses. The significant difference was found in the enamel changes between high or middle dose group and low dose group or control. The dentin morphology changed, some collagen fibre vanished and resistance to acid was reduced after irradiation with 50 Gy and 70 Gy.

CONCLUSIONSThe radiation reduced the resistance of teeth to the acid and increased the caries susceptibility.

Acids ; chemistry ; Dental Caries ; etiology ; Dental Caries Susceptibility ; Dental Enamel ; chemistry ; radiation effects ; ultrastructure ; Dentin ; chemistry ; radiation effects ; ultrastructure ; Humans ; Radiotherapy ; adverse effects