Objective: To investigate the effect of Helicobacter pylori (H. pylori)　on the synthesis of cyclooxygenase-1 (COX-1)　and cyclooxygenase-2 (COX-2) in gastric epithelial cells. Methods: a VacA(+) and CagA(+) international standard H. pylori line NCTC11637 and a human gastric epithelial carcinoma cell line BGC-823 were used. Western Blotting was applied to detect the synthesis of cyclooxygenase. Results: The content of COX-2 protein increased obviously after the cells were incubated with H. pylori sonicating extract for 1　h and the increase lasted for at least 6　h whereas the content of COX-1 protein did not change during the incubation with H. pylori extract. H. pylori lipopolysaccharide (LPS) had no effect on COX-2 synthesis. Conclusion: H. pylori stimulated the synthesis of COX-2 in BGC-823 cells and the effect was LPS-independent.

OBJECTIVETo study the impact of low-level lead exposure on neural cell adhesion molecule (NCAM) expression of primarily cultured hippocampal neurons.

METHODSWistar rats gestated at 18th day were anaesthetized and paunched to get the pups, the hippocampi of the pups were separated and the hippocampal neurons were primarily cultured. After co-cultivated with different dosage of PbCl(2), the NCAM expression of the neurons were tested with Western blotting at different culture time.

RESULTSNormally, the expression of NCAM at the 1st culture day was very low and its integral obsorbency density was 14; the climax expression time of NCAM of the cultured hippocampal neurons was 3rd to 5th cultured day, and their integral obsorbency density were 2 542 to 2 580; henceforth, the NCAM expression declined. NCAM expression was inhibited significantly by lead during the 2nd to 4th cultured day, and dose-response relationship was observed. The inhibition of lead weakened along with the cultured time prolonged, at 5th cultured day, it disappeared, and the NCAM expression of 10(-2), 10(-3) and 10(-4) mmol/L groups even exceeded the control groups. After that, the expression of NCAM in all groups began to decline, and the dose-response relationship of lead to the NCAM expression was observed again.

CONCLUSIONLow-level lead might significantly inhibit the NCAM expression of the primarily cultured Wistar rats' hippocampal neurons, and might delay the climax NCAM expression time.

Animals ; Animals, Newborn ; Cell Separation ; Cells, Cultured ; Dose-Response Relationship, Drug ; Female ; Hippocampus ; cytology ; drug effects ; metabolism ; Lead ; toxicity ; Neural Cell Adhesion Molecules ; biosynthesis ; genetics ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Wistar

In order to study the relationship between the expression of glutathione S-transferase (GST) in leukemic cells and the chemoresistance in patients with acute leukemia, the expressions of GST activity and GST mRNA were measured according to spectrophotometric assay based on the use of 1-choloro-2, 4-dinitro benzene and in situ hybridization. The results were studied in correlation with some clinical and pathological data. Results showed that: 1. There is no significant differences between activities of the enzyme with the different leukemia types according to the FAB classification. 2. GST activity and GST mRNA expression in the patients, both untreated and relapse, were (4.5 +/- 1.0) U, 33.3% and (7.9 +/- 15) U, 66.3% respectively. 3. In 56 patients, GST activity was 1.7 +/- 0.7, 5.9 +/- 2.0 and 9.3 +/- 1.7 U and GST mRNA expression was 13.3%, 29.7% and 76.6%, respectively, in CR, PR and NR groups. The lowest values of GST activity and GST mRNA expression were observed in those patients who achieved complete remission. The highest values of GST activity and GST mRNA expression were observed in those patients with no response to treatment. It was concluded that the expression of GST in patients with acute leukemia is closely related to the chemosensitivities clinically. Determinations of GST activity and GST mRNA are useful for predicting the chemosensitivities and the prognosis of the disease.
Adolescent ; Adult ; Aged ; Drug Resistance, Neoplasm ; Female ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Isoenzymes ; genetics ; metabolism ; K562 Cells ; Leukemia ; drug therapy ; enzymology ; genetics ; Leukemia, Lymphoid ; drug therapy ; enzymology ; genetics ; Leukemia, Monocytic, Acute ; drug therapy ; enzymology ; genetics ; Leukemia, Myeloid, Acute ; drug therapy ; enzymology ; genetics ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism

AIMTo investigate the effect of musk soluble components on the growth, the differentiation and the transfection efficiency of rat neural stem cell (NSC) in vitro.

METHODSThe growth and the differentiation of rat NSC were observed when musk soluble components were added into the culture medium of NSC. Meanwhile, the pEGFP-C1, which expressed the enhanced GFP protein, was transfected into the NSC by method of electro- transfection.

RESULTSWhen NSC was treated with musk soluble components, the neurites were outgrowth around NSC and attached to the plate, and the neural spheres were disassociated. The glia-like cells appeared at the concentration of 0.3 per thousand. When the concentration of musk soluble components was lower than 3 per thousand, the transformative cells could recover. Furthermore, the efficiency of transfection pEGFP into NSC was remarkably increased after the treatment with musk.

CONCLUSIONAfter the treatment of NSC with musk soluble components, the neural spheres were disassociated, and then attached to the plate. Musk soluble components could induce NSC differentiation into glia-like cells and improve the transfection efficiency of pEGFP-C1 in vitro.

Animals ; Brain ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; Fatty Acids, Monounsaturated ; chemistry ; Female ; Fetus ; Male ; Materia Medica ; pharmacology ; Neural Stem Cells ; cytology ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo investigate the safety and efficiency of the disposable circumcision suture device (DCSD) in the surgical treatment of phimosis and redundant prepuce.

METHODSWe randomly assigned 249 outpatients with phimosis or redundant prepuce to be treated with DCSD (n = 129) and by conventional circumcision (CC, n = 120), respectively. Then we compared the safety and efficiency of the two strategies.

RESULTSComparisons between DCSD and CC showed that the operation time was (4.02 +/- 0.69) vs (30.8 +/- 4.05) min, blood loss was (1.07 +/- 1.29) vs (8.72 +/- 2.15) ml, intraoperative pain score was 0.81 +/- 0.81 vs 2.42 +/- 1.15, 24-hour postoperative pain score was 1.84 +/- 1.02 vs 4.99 +/- 1.36, postoperative complication rate was 13. 95% (18/129) vs 9.17% (11/120), wound healing time was (13.99 +/- 9.06) vs (17.48 +/- 3.49) d, satisfaction with the penile appearance was 98.4% (127/129) vs 95% (109/120), and treatment cost was (2215.62 +/- 17.67) vs (576.47 + 15.58) Y RMB. DCSD exhibited obvious superiority over CC for shorter operation time, less blood loss, milder intraoperative pain, sooner wound healing, and better penile appearance, but it also had a higher rate of postoperative complications (P > 0.05) and involved more treatment cost than the latter (P < 0.05).

CONCLUSIONThe disposable circumcision suture device affords ideal clinical effects and therefore deserves clinical popularization.

Circumcision, Male ; instrumentation ; Disposable Equipment ; Follow-Up Studies ; Humans ; Male ; Phimosis ; surgery ; Surgical Staplers ; Treatment Outcome

AIMTo determine the effect of lysophosphatidylcholine (LPC) on the expression of vascular endothelial growth factor (VEGF) in human umbilical veins endothelial cell line (ECV304) and the inhibitory effect of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside (ST I) in vitro.

METHODSExposure to 2.5 mg x L(-1) LPC or LPC + ST I for 24 hours, VEGF protein was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, VEGF mRNA expression in ECV304 was examined by in situ hybridization. VEGF165 mRNA was examined by RT-PCR and Realtime RT-PCR.

RESULTSLPC upregulated VEGF protein and VEGF mRNA expression in the ECV304 cells. ST I was shown to markedly inhibit the LPC-induced increase of VEGF protein and VEGF165 mRNA (P < 0.001).

CONCLUSIONLPC can induce a strong expression of VEGF in ECV304 cells and ST I can inhibit it.

Cells, Cultured ; Endothelial Cells ; metabolism ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Lysophosphatidylcholines ; antagonists & inhibitors ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Stilbenes ; isolation & purification ; pharmacology ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo determine the role of hepatic oval cell in primary hepatocarcinogenesis using specific Y chromosome.

METHODSThe model of hepatic oval cell proliferation was established by feeding 30 male Wistar rats with 0.06% 3'3-diaminobenzidine (DAB) for 4 weeks. Another 40 female Wistar rats were equally randomized into two groups: control group and experimental group. Experimental group was inoculated with oval cells suspension from the prepared male rats under the hepatic amicula and fed with DAB for 14 weeks continuously to promote hepatocarcinogenesis. Control group was fed with DAB for 14 weeks. After primer fragments were worked out on the male rats' SRY genes using the software of primer design, DNAs were extracted from the hepatic carcinomas of the female rats of the two groups and then amplified with PCR. Electrophoretic analysis was performed on the products.

RESULTSAfter 14 weeks, primary hepatocarcinogenesis was found in the livers of all the 40 female rats. Electrophoresis showed the positive straps from the experimental group with the same length to the designed segments, whereas no positive straps were seen in the control group.

CONCLUSIONSThe hepatic oval cells can differentiate into hepatic cancer cells, which provides the evidence that the hepatocellular carcinoma has its source from the hepatic oval cells.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; physiopathology ; Cell Proliferation ; Cell Transformation, Neoplastic ; genetics ; Cells, Cultured ; Female ; Hepatocytes ; metabolism ; pathology ; Liver Neoplasms, Experimental ; genetics ; pathology ; physiopathology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Stem Cells ; metabolism ; pathology ; Y Chromosome

OBJECTIVETo study how CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms affect the prognosis of Chinese HIV-1 carrier.

METHODSEpidemiologic survey was done to the HIV-1 carriers who were found in Shenzhen area. PCR/RFLP technology was applied to analyze CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms of the HIV-1 carriers. The plasma virus load and CD4+ cell counting was assayed. The incubation period of some carriers was estimated. SPSS11.0 software was used to analyze the data.

RESULTSNo persons with CCR5delta32 and CCR5m303 mutation genotype were found from 189 HIV-1 carriers. SDF1-3'A allele frequency was 26.14% and CCR2-64I allele 19.82%. The carriers were divided into high virus load group (virus load < 20,000 copies/ml) and low virus load group (virus load > or =20,000 copies/ml). It was found by one-way ANOVA analysis on the logarithm of virus load that there was no significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.272). One-way ANOVA analysis on delitescence of some carriers showed that there was not significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.662). One-way ANOVA analysis on the logarithm of virus load showed that there was significant difference among SDF1-3'A wild genotype, cross bred genotype and pure mutation genotype (P=0.001).

CONCLUSIONCCR2-64I gene mutation may not significantly affect virus load of Chinese HIV-1 carriers, nor it affect the incubation period of HIV-1 carriers. SDF1-3'A gene mutation can decrease virus load, but it may not prolong the incubation period of HIV-1 carriers.

Acquired Immunodeficiency Syndrome ; genetics ; pathology ; virology ; Carrier State ; Chemokine CXCL12 ; genetics ; China ; Gene Frequency ; Genotype ; HIV-1 ; Heterozygote ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Prognosis ; Receptors, CCR5 ; genetics

In order to establish a new more rapid, safe and sensitive colorimetric assay for the proliferation of leukemic cells, MTS/pms has been developed. This automated colorimetric assay is based on the characteristic of viable and metabolically active leukemic cells to cleave MTS/pms into a water-soluble product whose optical density is determined at 492 nm by an automated microtiter-plate reader photometer. The results indicated that only active leukemic cells cleaved MTS/pms into product measured, and dead cells did not reduce MTS/pms. A linear relations hip were found between the viable cell number and optical density of MTS/pms cleaved by HL-60 and K562 cell (r = 0.963). Compared with MTT and INT assays, the reduced product of MTS/pms is water-soluble. It is concluded that MTS/pms colorimetric assay is more rapid, accurate and sensitive for the bioassay of proliferation of leukemic cells.
Cell Division ; Colorimetry ; methods ; Formazans ; metabolism ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; pathology ; Methylphenazonium Methosulfate ; metabolism ; Tetrazolium Salts ; metabolism ; Thiazoles ; metabolism

OBJECTIVETo investigate the effect of liposome-mediated glial cell line-derived neurotrophic factor (GDNF) gene transfer in vivo on spinal cord motoneurons after spinal cord injury (SCI) in adult rats.

METHODSSixty male Sprague-Dawley rats were divided equally into two groups: GDNF group and control group. The SCI model was established according to the method of Nystrom, and then the DC-Chol liposomes and recombinant plasmid pEGFP-GDNF cDNA complexes were injected into the injured spinal cord. The expression of GDNF cDNA 1 week after injection was detected by RT-PCR and fluorescence microscope. We observed the remaining motoneurons in the anterior horn and the changes of cholinesterase (CHE) and acid phosphatase (ACP) activity using Nissl and enzyme histochemistry staining. The locomotion function of hind limbs of rats was evaluated using inclined plane test and BBB locomotor scale.

RESULTSRT-PCR and fluorescence observation confirmed the presence of expression of GDNF cDNA 1 week and 4 weeks after injection. At 1, 2, 4 weeks after SCI, the number of motoneurons in the anterior horn in GDNF group (20.4+/-3.2, 21.7+/-3.6, 22.5+/-3.4) was more than that in control group (16.8+/-2.8, 17.3+/-2.7, 18.2+/-3.2, P<0.05). At 1, 2 weeks after SCI, the mean gray of the CHE-stained spinal motoneurons in GDNF group (74.2+/-25.8, 98.7+/-31.6) was less than that in control group (98.5+/-32.2, 134.6+/-45.2, P<0.01), and the mean gray of ACP in GDNF group (84.5+/-32.6, 79.5+/-28.4) was more than that in control group (61.2+/-24.9, 52.6+/-19.9, P<0.01). The locomotion functional scales in GDNF group were higher than that in control group within 1 to 4 weeks after SCI (P<0.05).

CONCLUSIONSGDNF gene transfer in vivo can protect motoneurons from death and degeneration induced by incomplete spinal cord injury as well as enhance locomotion functional restoration of hind limbs. These results suggest that liposome-mediated delivery of GDNF cDNA might be a practical method for treating traumatic spinal cord injury.

Animals ; Disease Models, Animal ; Gene Transfer Techniques ; Glial Cell Line-Derived Neurotrophic Factor ; Injections, Intralesional ; Liposomes ; Locomotion ; physiology ; Male ; Motor Neurons ; drug effects ; Nerve Growth Factors ; pharmacology ; Nerve Regeneration ; physiology ; Neuroprotective Agents ; pharmacology ; Primary Prevention ; methods ; Probability ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Reference Values ; Reverse Transcriptase Polymerase Chain Reaction ; Spinal Cord Injuries ; pathology ; prevention & control ; therapy