Arsenicals ; pharmacology ; DNA Methylation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Membrane Proteins ; genetics ; metabolism ; Oxides ; pharmacology

Adolescent ; Child ; Diagnosis, Differential ; Dystonic Disorders ; diagnosis ; drug therapy ; physiopathology ; Female ; Humans ; Male

BACKGROUND:The high level of matrix metalloproteinase 9 (MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fibroblasts and affect wound healing is stillunclear. The present study aimed to observe changes in the biological behaviors of rat dermal fibroblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot. METHODS:A cellmodel of skin fibroblast with high expression of MMP9 was established by co-culture of high glucose (22.0 mmol/L) and homocysteine (100 μmol/L). A control group was incubated with normal glucose (5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwellwere used to detect cellproliferation, viability, collagen (hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically significant. RESULTS:The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group (7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively,P<0.01). The proportion of S-phase cells, proliferation index, cellviability, collagen (hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group (P<0.01). CONCLUSION:Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fibroblasts.

OBJECTIVETo explore the dynamic changes of blood sugar and body's signs in streptozotocin diabetic animal models.

METHODSRat and mouse diabetic models were established by a single intraperitoneal (ip) injection and 5-day successive ip injections of streptozotocin respectively. Blood sugar levels were measured. The food consumption index (consumption of food/body weight) and the water consumption index (consumption of water/body weight) were calculated.

RESULTSSixty five point zero percent male rats received streptozotocin, 60 mg/kg ip, developed diabetes mellitus. The blood sugar remained in high level between the 15th day and the 25th day after injection, and it began to decline afterwards. By 5-day ip injections of streptozotocin, 40 mg/kg daily, 90.0% male mice developed diabetes mellitus. Dynamic changes of blood sugar of diabetic mouse were similar to those of rats, except that the blood sugar of mice did not decline as obvious as that of rats. The changes of water consumption index were in best fit with the changes of blood sugar in both models, with correlation index r>0.970.

CONCLUSIONThe blood sugar of diabetic animal model stayed in high level from the 15th day to the 25th day after the beginning of injection. And the period is suitable for observing effect of anti-diabetic drugs. The water consumption index can reflect the blood sugar levels of diabetes animals.

Animals ; Blood Glucose ; analysis ; Body Weight ; physiology ; Diabetes Mellitus, Experimental ; blood ; chemically induced ; physiopathology ; Disease Models, Animal ; Drinking ; physiology ; Eating ; physiology ; Male ; Mice ; Rats ; Rats, Wistar ; Streptozocin

OBJECTIVETo investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant.

METHODSDCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method.

RESULTSDCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg.

CONCLUSIONHuman dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.

Adolescent ; Adult ; Antigen Presentation ; Cells, Cultured ; Child ; Dendritic Cells ; drug effects ; immunology ; Female ; Hepatitis B Surface Antigens ; pharmacology ; Hepatitis B, Chronic ; immunology ; Humans ; Lymphocyte Activation ; Male ; Middle Aged

Aim The effect of tetrandrine on TGF-?1 mRNA expression in glomerulosclerosis rat was observed. Methods The rats were randomly divided into four groups, such as the normal control group (sham operative rat), glomerulosclerosis model group,tetrandrine group and amlodipine group. The expression of TGF-?1 mRNA was analyzed by Northern blot hybridization. Results The expressions of TGF-?1 mRNA in two treating groups were much lower than untreated model group. There were no difference between these two treating groups. Conclusion Tetrandrine can decrease the expression of TGF-?1 mRNA in glomerulosclerosis rat induced by unilateral renctomy plus adriamycin.

OBJECTIVE: To study DNA quantification and STR typing of samples pre-treated with pyramidon. METHODS: The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. RESULTS: In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. CONCLUSION Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology* ; Blood Stains ; DNA/isolation & purification* ; DNA Fingerprinting ; Forensic Medicine ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results ; Specimen Handling

OBJECTIVETo evaluate clinical outcomes of a group of adolescent idiopathic scoliosis (AIS) patients undergoing posterior pedicle screw-only instrumentations.

METHODSBetween April 2002 and July 2006, 121 AIS patients (93 female and 28 male, average age at operation was 15.5 years which ranged from 10 to 20 years) received posterior pedicle screw-only instrumentation and fusion. All the patients were evaluated by the various-parameters measured in X-ray films before and after surgery, including Cobb angle on coronal plane, Cobb angle on sagittal plane, clavicle angle and shoulder height difference, lowest instrumented vertebrae (LIV) angulation, proximal junction kyphotic angle, the distances of central sacral vertical line (CSVL) to the LIV, to the apical vertebra and to the C(7) plumb line respectively. Complications were followed.

RESULTSAn average of (11.0 + or - 1.5) levels was fused. An average coronal correction of proximal thoracic curve was 41.8%, of thoracic curve was 70.8%, of thoracolumbar/lumbar curves was 74.0%. No significant change was found in sagittal alignment. Shoulder balance and apex vertebral to central sacral line were restored well. There were no pseudoarthroses and loss of correction during the follow-ups. One adding-on, 4 proximal thoracic decompensation and 15 proximal junction kyphosis were found during the follow-ups.

CONCLUSIONPosterior pedicle screw-only instrumentation and fusion has excellent radiographic and clinical results with minimal complications in the surgical treatment of AIS.

Adolescent ; Bone Screws ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Retrospective Studies ; Scoliosis ; surgery ; Spinal Fusion ; instrumentation ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo compare the outcomes of the three microsurgical strategies, inguinal high ligation (IHL), retroperitoneal high ligation (RHL) and low ligation (LL) of internal spermatic veins, in the treatment of varicocele.

METHODSWe retrospectively analyzed 120 cases of varicocele, which were equally divided into groups I , II and III to be treated by IHL, RHL and LL of internal spermatic veins, respectively. We compared the operation times, post-operative complications, recurrence rates and results of pre- and post-operation semen analysis among the three groups.

RESULTSThe mean operation time was significantly longer in group III ( [55 +/- 6 ] min) than in I ([35 +/- 10] min) and II ([42 +/- 12] min) (P<0.05), while the rate of post-operative complications remarkably higher in group I (4 cases of hydrocele [10% ] and 3 cases of epididymitis [7.5%]) than in II (2 cases of hydrocele [5%] and 2 cases of epididymitis [5%]) and III (1 case of hydrocele [2.5%] and 1 case of epididymitis [2.5%]) (P<0.05). Six to 12 (mean 9) months of follow-up visit found 2 cases (5% ) of recurrence in group I, 1 case (2.5%) in group II and none in group III, with no statistically significant difference among the three groups (P>0.05). At 12 months after surgery, group III showed significantly higher sperm concentration, grade a +b sperm and the sperm motility than the other two (P<0.05), but no significant differences were observed in these parameters among the three groups preoperatively.

CONCLUSIONAs a microsurgical approach to the treatment of varicocele, LL is better than IHL and RHL of internal spermatic veins in improving the seminal parameters of the patients.

Humans ; Ligation ; Male ; Microsurgery ; adverse effects ; methods ; Retrospective Studies ; Treatment Outcome ; Urologic Surgical Procedures, Male ; adverse effects ; methods ; Varicocele ; surgery

OBJECTIVETo investigate the role of Apaf-1 gene promoter methylation and apoptosis inhibitor protein Apollon in pathogenesis of acute leukemia (AL) and their clinical significance.

METHODSMethylation specific PCR (MSP) was used to detect the methylation status of Apaf-1 gene promoter in 53 AL patients (28 AML, 10 ALL and 15 relapsed) and 10 healthy or nonmalignant blood diseases patients as control. RT-PCR was used to detect the expression levels of Apaf-1 mRNA and immunocytochemistry to detect the expression levels of Apollon protein.

RESULTSThe abnromal methylation of Apaf-1 gene promotor in AL was 18/53(33.9%). No Apaf-1 mRNA was detected in methylation positive patients. Only one case in healthy and nonmalignant individuals was deletion of Apaf-1 mRNA expression without abnormal methylation. The positive methylation rate in AL bone marrow mononuclear cells was significantly higher than that in controls (P < 0.05). The expressin levels of Apollon protein in AL patients was higher than that in control (P < 0.05). The positive methylation ratio and Apollon protein level were higher in white blood cell count > 10 × 10(9)/L than in ≤ 10 × 10(9)/L (P < 0.05). There is a positive correlaiton between positive methylation ratio and Apollon protein expression in AL patients.

CONCLUSIONAbnormal methylation of Apaf-1 gene promotor and high expression of Apollon might involved in leukemogenesis.

Acute Disease ; Adult ; Apoptosis ; DNA Methylation ; Humans ; Leukemia ; genetics ; Promoter Regions, Genetic ; RNA, Messenger ; genetics