OBJECTIVETo investigate the collagen constitution of hyperplastic scar (HS) in different ages and the change of relative factors.

METHODSThirty cases with HS were divided into two groups according to patients' age: group 1 (1 - 19 years, A) and group 2 (20 - 50 years, B). The normal skin (NS) from corresponding age of volunteers was employed as control group. The changes in TGFbeta1, collagenase (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1beta) and the collagen ratio were observed by means of in situ hybridization technique and SABC (Strept-Avidin-Biotin complex) immunohistochemistry and image analysis.

RESULTSThe ratio of type I to type III collagen in A group was 6.48 in average and 3.76 in B group, but there was no evident difference in the ratio during the disease process in both groups. The expression of TGFbeta1 in A group was much higher than that in B group (P < 0.01). The TIMP-1 mRNA expression showed no difference among all age groups in HS patients, but it was much higher than that in NS group. The MMP-1 expression was evidently lower than TIMP-1 expression, and there was no difference in MMP-1 expression compared with NS group.

CONCLUSION(1) The TGFbeta1 expression in HS patients was negatively correlated with age, and the increased expression of TGFbeta1 produced an increase ratio of type I to type III collagen. (2) High level expression of TIMP-1 led to the formation of HS by inhibiting MMP-1 expression, and the expression was not related to age.

Adolescent ; Adult ; Age Factors ; Burns ; metabolism ; pathology ; Child ; Child, Preschool ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Female ; Humans ; Infant ; Male ; Matrix Metalloproteinase 1 ; biosynthesis ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; Transforming Growth Factor beta1 ; biosynthesis ; Young Adult

OBJECTIVETo evaluate whether or not administration of folic acid and resveratrol have preventive effects on cleft palate formation as well as the comparison of the two drugs' s effects.

METHODSPregnant mice were randomly divided into 9 groups, with 8 mice in each group. The TCDD group mice were dosed with TCDD 28 microg/kg body weight on gestation day 10 (GD 10) animals in folic acid group were respectively dosed with folic acid 15, 10, 5 mg/kg and TCDD 28 microg/kg; resveratrol treated mice were divided into 3 groups: resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13 in resveratrol (GD8-13 ) group; resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13, followed hy an oral administered with TCDD on GD10 in resveratrol (GD8-13) + TCDD group; resveratrol 50mg/kg and TCDD 28 microg/kg were used by gavage administration at GD10 in resveratrol (GD10) + TCDD group. Control mice were treated with the same volume of water for 6 consecutive days from GD8 to GD13 and were given a single dose of corn oil on GD10. The pregnant mice weight and embryos, the number of live, cleft palate, dead and resorption fetal mice were recorded on GD 17.5. The coronal sections of the fetal mice heads were prepared at GD 17.5 and observed by microscopy.

RESULTSTotal frequency of clefts was 92.86% in TCDD group, 84.00% (15 mg), 73.08% (10 mg), 84.00% (5 mg) in folic acid + TCDD groups, 0% in resveratrol (GD10) group, 74.51% (GD10), 57.78% (GD8-13) in resveratrol + TCDD groups. The frequency of cleft was 0% in the control group. Compared with the control and the TCDD groups, there were significant differences in the number of live, dead and resorption fetal mice in TCCD + resveratrol (GD8-13) group (P < 0.05). No significant differences in embryonic weight, live fetuses weight, the number of live, dead and resorption fetal mice were found in the other groups (P > 0.05).

CONCLUSIONTest dose of folic acid and resveratrol both had certain antagonistic effect on cleft palate in mice induced by TCDD, with folic acid 10 mg/kg, resveratrol 50 mg/kg GD8-13 doses having stronger antagonistic action. Effects of both the two drugs have no significant difference, but resveratrol (50 mg/kg, GD8-13) significantly affects the fetal mice's growth and development under TCDD exposure in utero.

Abnormalities, Drug-Induced ; prevention & control ; Animals ; Cleft Palate ; chemically induced ; prevention & control ; Female ; Fetus ; Folic Acid ; administration & dosage ; pharmacology ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; antagonists & inhibitors ; Pregnancy ; Random Allocation ; Stilbenes ; administration & dosage ; pharmacology ; Teratogens

BACKGROUNDIn response to the inflammatory reaction, circulating leukocytes aggregate and adhere to the endothelial cells and eventually pervade into tissues, resulting in cell damage. This study was to detect the inflammatory reactions in mouse focal cerebral ischemia and their distinct characteristics in the ischemic basal ganglia and surrounding cortex.

METHODSMice were subjected to permanent occlusion of the left middle cerebral artery (MCAO) by introducing a suture for 2 to 120 hours. The expression of intercellular adhesion molecule 1 (ICAM-1) and Mac-1 was determined immunohistochemically. The myeloperoxidase (MPO) activity of the ischemic regions was measured.

RESULTSFour hours after MCAO, the number of ICAM-1 positive vessels in the ischemic basal ganglia increased (9.2 +/- 2.8 per mm(2)), peaked at 48 hours (29.6 +/- 4.8 per mm(2)), and decreased after 72 hours. In the ischemic cortex, the number increased rapidly 4 hours after MCAO (19.4 +/- 6.1 per mm(2)), peaked at 48 hours (44.4 +/- 16.8 per mm(2)), and declined after 72 hours. Mac-1 positive cells were seen in the ischemic basal ganglia (3.4 +/- 1.2 per mm(2)) 12 hours after MCAO, peaked after 48 hours (20.2 +/- 6.3 per mm(2)), and decreased after 72 hours. In the ischemic cortex, however, the number increased 4 hours after MCAO (4.3 +/- 1.7 per mm(2)), peaked after 48 hours (20.9 +/- 8.4 per mm(2)), and remained high at 120 hours. The MPO activity increased in the ischemic basal ganglia 12 hours after MCAO (0.111 +/- 0.023 U/g), peaked after 24 hours (0.194 +/- 0.059 U/g), and decreased after 72 hours. In the ischemic cortex, the MPO activity increased 12 hours after MCAO (0.110 +/- 0.032 U/g), peaked after 24 hours (0.210 +/- 0.067 U/g), and remained elevated at 120 hours.

CONCLUSIONSThe increased expression of ICAM-1 in the ischemic brain of mouse in the early phase of MCAO followed by the over-expression of Mac-1 and the increased MPO activity suggests that focal ischemia leads to early onset of inflammation. The inflammatory response is more persistent and intensive in the ischemic cortex than in the ischemic basal ganglia.

Animals ; Basal Ganglia ; blood supply ; Brain Chemistry ; Brain Ischemia ; metabolism ; pathology ; Cerebral Cortex ; blood supply ; Cerebrovascular Circulation ; Inflammation ; etiology ; Intercellular Adhesion Molecule-1 ; analysis ; Macrophage-1 Antigen ; analysis ; Male ; Mice ; Middle Cerebral Artery ; Peroxidase ; analysis

AIMNucleoside analogues have become the most promising candidates of anti-HBV drugs. In this study, beta-L-D4A was synthesized and explored its inhibitiory action against hepatitis B virus (HBV) in 2. 2. 15 cells derived from HepG2 cells transfected with HBV genome.

METHODSbeta-L-D4A was stereo-controlled synthesized from D-glutamic acid, and the structure was identified by IR, 1H NMR and MS. 2. 2. 15 Cells were placed at a density of 5 x 10(4) per well in 12-well tissue culture plates, and treated with various concentrations of beta-L-D4A for 6 days. At the end, medium was processed to obtain virions by a polyethlene glycol precipitation method. At the same time, intracellular DNA was also extracted and digested with Hind III. Both of the above DNA were subjected to Southern blot, hybridized with a 32P-labeled HBV probe and autoradiographed. The intensity of the autoradiographic bands was quantitated by densitometric scans of computer and EC50 was calculated. 2. 2. 15 cells were also seeded in 24-well tissue culture plates, and cytotoxicity with different concentrations was examined by MTT method. IC50 was calculated.

RESULTSThe synthesized compound structure conformed with beta-L-D4A; Autoradiographic bands showed similar for supernatant and intracellular HBV DNA. Episomal HBV DNA was inhibited in a dose-dependent manner. EC50 0.2 micromol x L(-1). The experiment of cytotoxicity gained IC50 200 micromol x L(-10.

CONCLUSIONbeta-L-D4A has been synthesized successfully. beta-L-D4A possessed potent inhibitory effect on replication of HBV in vitro with low cytotoxicity, TI value was 1 000. It is expected to be developed clinically into a new anti-HBV drug.

Antiviral Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Replication ; drug effects ; DNA, Viral ; drug effects ; Dideoxyadenosine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; physiology ; Humans ; Liver Neoplasms ; pathology ; Transfection ; Virus Replication ; drug effects

OBJECTIVETo investigate the method for simultaneous correction of nasal deformity and unilateral cleft lip so as to decrease the secondary operation for the deformity.

METHODSThe Millard procedure (or Millard plus triangle flap insertion) was used to repair the unilateral cleft lip. Through the incisions, the greater alar and nasalis were repositioned to the normal anatomical positions. The deviated septum and columella were corrected by cutting the abnormal attachment of the orbicular muscle of mouth to the anterior nasal spine. The mattress sutures through the tip of the columella and ala nasi helped to recover the shape of the nostril.

RESULTS108 patients were treated with this method. They aged from one month to 19 years, included 30 with second degree cleft lip and 78 with third degree cleft lip. The follow-up for as long as 3 years showed satisfactory results.

CONCLUSIONSThis technique can eliminate the severe cleft nasal deformity and elevate the displaced alar cartilage at the time of lip repair without interference with nasal growth. It is recommended for the treatment of unilateral cleft lip with severe nasal deformity.

Abnormalities, Multiple ; surgery ; Adolescent ; Child ; Child, Preschool ; Cleft Lip ; surgery ; Female ; Humans ; Infant ; Male ; Nose ; abnormalities ; surgery ; Reconstructive Surgical Procedures ; Rhinoplasty ; methods ; Young Adult

OBJECTIVETo explore a method to repair nasal side mucosa of wide incomplete cleft palate and reduce the tension of wound by using oral mucosa flap in the top of fissure.

METHODS27 cases of wide incomplete cleft palatal were included in the study. On the basis of two-flap palatoplasty, the triangular oral mucosa flap in the top of fissure was turned and sewed with side mucosa to repair nasal side mucosa of wide palatal cleft.

RESULTSWithout postoperative active bleeding, airway obstruction and wound infection, 27 cases had been repaired satisfactorily by this procedure. 1-3 months followed up demonstrated that all the wounds healed well without wound dehiscence or fistulas and the scars in the palate were not severe.

CONCLUSIONUsing oral mucosa flap in the top of fissure to repair nasal side mucosa of wide palatal cleft can get a reduced tension and correspondingly increase the width of mucoperiosteal flaps so as to decrease incidence rate of palatal fistulas and reduce formation of scars.

Cleft Palate ; Female ; Humans ; Mouth Mucosa ; Nasal Mucosa ; Reconstructive Surgical Procedures ; Surgical Flaps

OBJECTIVETo evaluate the effect of Helicobacter pylor (Hp) eradication in children with acute idiopathic thrombocytopenic purpura (ITP).

METHODSNinety-three children with acute ITP and Hp infection were divided into two groups and treated with prednisone and Hp eradication (group A, 51 cases) or with prednisone without Hp eradication (group B, 42 cases).

RESULTSThe Hp eradication rate was 94.1% in group A. No difference was found in the therapeutic effects on IPT between the two groups, but the recurrence rate in one year in group A was significantly lower than that in group B.

CONCLUSIONNHp eradication does not obviously enhance the therapeutic effect on childhood acute ITP, but can decrease the relapse rate in one year. HP eradication therapy is recommended in children with acute ITP and Hp infection.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Therapy, Combination ; Female ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; drug effects ; immunology ; Humans ; Male ; Prednisone ; therapeutic use ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; microbiology

OBJECTIVETo study the role of extracellular signal-regulated kinase (ERK) in cerebral ischemia and the mechanism of protective effects of U0126 (1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio] butadiene) on ischemic brain.

METHODSMice underwent left middle cerebral artery occlusion (MCAO) by introducing a suture in the lumen. U0126 was injected intravenously through the internal jugular vein. The immuno-activity of phosphorylated ERK1/2 (pERK1/2), phosphorylated mitogen activated protein kinase kinase (pMEK), and phosphorylated Elk-1 (pElk-1) was assessed by Western blot analysis and immunohistochemistry. Interleukin (IL)-1beta mRNA level was measured by ribonuclease protection assay.

RESULTSPhosphorylated ERK1/2 in 2 hours MCAO mice was down-regulated after intravenous injection of U0126. The inhibition was dose dependent and treatment time related. pMEK and pElk-1 were also reduced in a similar fashion after U0126 treatment. IL-1beta mRNA increased after 1 and 2 hours of MCAO. After injection of U0126, it was down-regulated during 1 to 4 hours after MCAO.

CONCLUSIONIntravenous administration of the MEK inhibitor U0126 inhibits pMEK, pERK1/2, and pElk-1 up-regulation induced by cerebral ischemia. The protective effect of U0126 against ischemic injury is probably resulted from the reduction of IL-1beta mRNA via the inhibition of ERK pathway.

Animals ; Butadienes ; pharmacology ; DNA-Binding Proteins ; metabolism ; Enzyme Inhibitors ; pharmacology ; Infarction, Middle Cerebral Artery ; metabolism ; Interleukin-1 ; biosynthesis ; genetics ; Male ; Mice ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase Kinases ; antagonists & inhibitors ; metabolism ; Nitriles ; pharmacology ; Phosphorylation ; Proto-Oncogene Proteins ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Signal Transduction ; Transcription Factors ; metabolism ; ets-Domain Protein Elk-1

Estrogen has anti-colorectal cancer effects which are thought to be mediated by mismatch repair gene (MMR) activity. Estrogen receptor (ER) expression is associated with microRNA (miRNA) expression in ER-positive tumors. However, studies of direct link between estrogen (especially estradiol E2), miRNA expression, and MMR in colorectal cancer (CRC) have not been done. In this study, we first evaluated the effects of estradiol (E2) and its antagonist ICI182,780 on the expression of miRNAs (miR-31, miR-155 and miR-135b) using COLO205, SW480 and MCF-7 cell lines, followed by examining the association of tissue miRNA expression and serum E2 levels using samples collected from 18 colorectal cancer patients. E2 inhibited the expressions of miRNAs in COLO205 cells, which could be reversed by E2 antagonist ICI 182.780. The expression of miR-135b was inversely correlated with serum E2 level and ER-beta mRNA expression in CRC patients' cancer tissues. There were significant correlations between serum E2 level and expression of ER-beta, miR-135b, and MMR in colon cancer tissue. This study suggests that the effects of estrogen on MMR function may be related to regulating miRNA expression via ER-beta, which may be the basis for the anti-cancer effect in colorectal cells.
Adaptor Proteins, Signal Transducing/genetics/metabolism ; Adult ; Aged ; Cell Line, Tumor ; Colorectal Neoplasms/*genetics/metabolism ; DNA Mismatch Repair/*genetics ; Estradiol/analogs & derivatives/blood/*pharmacology ; Estrogen Antagonists/pharmacology ; Estrogen Receptor beta/genetics/*metabolism ; Female ; *Gene Expression Regulation, Neoplastic ; Humans ; Male ; MicroRNAs/genetics/*metabolism ; Middle Aged ; MutS Homolog 2 Protein/genetics/metabolism ; Nuclear Proteins/genetics/metabolism ; RNA, Messenger/biosynthesis

BACKGROUNDTreponema pallidum (T. pallidum) subsp. pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.

METHODST. pallidum subsp. pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.

RESULTSRecombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.

CONCLUSIONSThe recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.

Animals ; Antigens, Bacterial ; genetics ; immunology ; Bacterial Proteins ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Membrane Proteins ; genetics ; immunology ; Polymerase Chain Reaction ; Rabbits ; Syphilis ; immunology ; microbiology ; Treponema pallidum ; immunology ; metabolism