BACKGROUND: Previous studies have confirmed that in the animal models of articular cartilage defects and osteoarthritis, the chondrocytes can overexpress the matrix metal oproteinases. Various abnormal stimuli are likely to break the balance between matrix metal oproteinase and tissue inhibitor of metal oproteinase, thus leading to degeneration of extracel ular matrix of articular cartilage, as wel as the decline and offset of cartilage chondrocytes. OBJECTIVE: To observe the effect of cyclic tensile strain on the expression of matrix metal oproteinases during the repairing process of rabbit articular cartilage defects. METHODS: The animal models of articular cartilage defects were established, and chondrocytes were separated for culture at 10 weeks after operation. The chondrocytes on the non-surgical side were considered as the normal group, and the chondrocytes on the surgical side were randomly divided into high cyclic tensile strain group, low cyclic tensile strains group and control group, and the load amplitude was sin10%. Then 0.1, 1.0 and 0 Hz cyclic tensile strains were loaded respectively. The expressions of matrix metal oproteinases 2, 3, 9 and 13 in each group were detected with reverse transcription-PCR at 24, 48 hours, 1, 2 and 4 weeks after loading cyclic tensile strain. RESULTS AND CONCLUSION: There were significant differences in the expressions of matrix metal oproteinases 2, 3, 9 and 13 at 24 hours after loading cyclic tensile strain between the normal group and the control group (P < 0.05); and there were significant differences in the expressions between the high cyclic tensile strain group and the low cyclic tensile strain group at 1, 2 and 4 weeks after loading cyclic tensile strain (P < 0.05).At the same time, the expressions of matrix metal oproteinases 2, 3, 9 and 13 in the low cyclic tensile strain group were continued to decline, and there were significant differences in the expressions after loading cyclic tensile strain for 24 hours and 4 weeks (P < 0.05). The results indicate that mechanical load can affect the expression of matrix metal oproteinases in the healing process of rabbit articular cartilage defects. In the cel ular and molecular level, the incidence and development of pathological articular cartilage defect and stress should affect each other.

Aim To investgate the mechnism through which ginkgolides affect learning and memory capabilities of the Alzheimers disease-like rats. Methods Okadaic acid(OA)was injected into the CA1 region of the rat hippocampus and the rats were gavaged with ginkgolides. The learning and memory abilities of the rats were assessed through Morris water maze behavioral test, and the expressions of nicotinic acetylcholine receptors and ChAT were observed by Western blotting and immunohistochemistry, respectively.Results Compared with the control rats, the capabilities of learning and memory were lowered significantly(P

OBJECTIVETo explore the technique and therapeutic effect of bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral shaft fractures in young children. Methods:From June 2006 to June 2012,45 young children with femoral shaft fractures were treated by bridge wire splint fixation with ankle dorsiflexion,which was designed according to arch bridge mechanical principle and structure. There were 31 males and 14 females with an average age of 3.2 years old ranging from 8 months to 5.5 years old; 14 cases were upper 1/3 femoral fractures,26 cases were middle 1/3 femoral fractures,5 cases were lower 1/3 femoral fractures; 20 cases were transverse fractures, 14 cases were oblique fractures,6 cases were spiral frac- tures, and 5 cases were comminuted fractures. X-ray, follow-up imaging changes,clinical curative effect and complications were assessed.

RESULTSForty-five patients were followed up for 6 to 21 months (averaged 12 months). All fractures were reached clinical bone healing after 5 to 7 weeks (averaged 6 weeks) fixation. Seven cases appearred limb soft tissue complications, including buttocks bedsore,dorsal foot and Achilles tendon epidermal necrosis, and healed after dressing and removal of external fixation. During follow-up,the original overlap angle and lateral displacement were remodeled, and limbs were restored to the normal line of force and bone structure. According to Flynn standard, 35 cases got excellent results, 8 cases good, 2 cases fair.

CONCLUSIONThe bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral. shaft fractures in young children (less than 6 years old) is safe,feasible, simple,and has raliable effect, which can be applied in primary hospitals.

Ankle ; surgery ; Bone Wires ; Child ; Child, Preschool ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; instrumentation ; Humans ; Infant ; Male ; Treatment Outcome

Antibiotic Prophylaxis ; Child ; Child, Preschool ; Cross Infection ; prevention & control ; Female ; Heart Defects, Congenital ; therapy ; Humans ; Infant ; Male

OBJECTIVETo study gene mutations in four pedigrees with methymalonic aciduria, as well as the feasibility of prenatal diagnosis of methymalonic aciduria.

METHODSHigh-throughput sequencing was performed for related genes in the peripheral blood of children or parents who were diagnosed with methymalonic aciduria to identify the loci with mutations. Then amplification primers were designed for each locus, and PCR and direct sequencing were performed to validate the sequencing in the first generation in the four pedigrees. Whether the mutations were pathogenic were determined with reference to literature review and medical history. In the pedigrees 1, 3, and 4, ultrasound-guided chorionic villi biopsy was performed at weeks 11-13 of pregnancy to perform early prenatal diagnosis.

RESULTSIn pedigree 1, c.656A>T and c.729-730insTT heterozygous mutations in the MUT gene were detected in the proband's father and mother, respectively. Early prenatal diagnosis showed c.656A>T and c.729-730insTT double heterozygous mutations in the fetus. The couple decided to terminate pregnancy. In pedigree 2, c.1106G>A and c.755-756insA double heterozygous mutations in the MUT gene were detected in the proband. c.1106G>A came from the father and c.755-756insA came from the mother. In pedigree 3, c.217C>T and c.609G>A double heterozygous mutations in the MMACHC gene were detected in the proband. c.217C>T came from the father and c.609G>A came from the mother. Prenatal diagnosis showed c.609G>A heterozygous mutation in the fetus. The baby was successfully delivered, and the result of umbilical cord blood testing was consistent with the prenatal diagnosis. In pedigree 4, c.609G>A and c.567dupT double heterozygous mutations in the MMACHC gene were detected in the proband. c.609G>A came from the father and c.567dupT came from the mother. Prenatal diagnosis showed c.567dupT heterozygous mutation in the fetus. The baby was successfully delivered, and the result of umbilical cord blood testing was consistent with the prenatal diagnosis.

CONCLUSIONSIdentification of gene mutations helps with prenatal diagnosis in pedigrees with methymalonic aciduria.

Amino Acid Metabolism, Inborn Errors ; diagnosis ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Male ; Mutation ; Pedigree ; Prenatal Diagnosis

Hepacivirus ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viremia ; virology

OBJECTIVETo observe the effect of Qihuang Decoction (QHD) on mRNA expression of apoptosis genes Bcl-2, Bax, and signal transduction molecules Caspase-3, 9 in intestinal mucosa epithelium of ischemia/ reperfusion (I/R) injured rats.

METHODSForty Wistar rats were randomized equally into 4 groups, the control group, the model group, the glutamine group, and the QHD group. Rats in the latter two groups were gastric infused with glutamine and QHD respectively for 3 days, but saline was infused instead to rats in the control group and model group. After then, except those in the control group intervened only by sham operation, rats were made into I/R injured model by 45 min occlusion of superior mesenteric artery followed by 1 h reperfusion. Immediately after modeling, mRNA expressions of Bcl-2, Bax, Caspase-3, and Caspase-9 in intestinal mucosa epithelium of rats were detected by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSCompared with the control group, mRNA expressions of Bcl-2, Bax, Caspase-3 and Caspase-9 were higher in the other three groups (P < 0.05). Compared with the model group, Bcl-2 mRNA expression was higher, while the expressions of the other three indices were lower in both the glutamine group and the QHD group (P < 0.05); and comparisons between the glutamine group and the QHD group showed a more depressed Bax mRNA expression (0.281 +/- 0.087 vs 0.350 +/- 0.053) and higher Bcl-2/Bax ratio (1.648 vs 1. 374) in the QHD group.

CONCLUSIONSQHD can reduce the I/R injury in the intestinal mucosa epithelium by inhibiting the cell apoptosis. The mechanism may be correlated with increased Bcl-2 mRNA expressions and decreased mRNA expressions of Bax, Caspase-3 and Caspase-9.

Animals ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelium ; metabolism ; Intestinal Mucosa ; drug effects ; metabolism ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo observe the effect of Qihuang Decoction (QHD) on epithelial cell apoptosis of ischemia-reperfusion (I/R) injured intestinal mucosa in rat.

METHODSThe I/R injured intestinal mucosa rat model was established by clamping superior mesenteric artery (SMA) for 45 min and reperfusing for 60 min. The pathomorphological Changes and epithelial cell apoptosis in the injured intestinal mucosa were observed and compared among groups: the sham-operated group (A), the model group (B), the glutamine treated group (C) and the QHD treated group (D).

RESULTSpathomorphological examination showed that in group A, the intestinal villus was intact; in group B, the intestinal subepithelial space were dilated, and showed evident cleavage between the epithelial top and the lamina propria with bare capillaries, bleeding and ulceration; in group C and D, the above-mentioned pathomorphological changes were alleviated to some extents, appeared only in part of the villa, and the alleviation was more significant in group D than in group C. Chiu's scoring showed that the lowest score (zero) presented in group A and the highest presented in group B; scores in group C and D was significantly lower than that in group B (P < 0.05), but showed insignificant difference between the two groups (P > 0.05). Epithelial cell apoptosis detection showed that the least apoptosis rate presented in group A, and the highest in the group B; while in the group C, it lied between group A and B (all P < 0.05), and showed no statistical significance to group D (P > 0.05), though appeared a lowering trend.

CONCLUSIONQHD could reduce the I/R injured intestinal epithelial mucosa, and its protective mechanism may be related to the inhibition on apoptosis of intestinal mucosal epithelial cells.

Animals ; Apoptosis ; drug effects ; Astragalus Plant ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; Intestinal Mucosa ; blood supply ; drug effects ; pathology ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology

Objective To evaluate the clinical effect in the treatment of the young patients(≤45 years old) with acute myocardial infarction(AMI)underwent emergent percutaneous transluminal coronary angioplasty(PTCA) combined with Lipo-PGE_1.Methods 39 patients with AMI(paroxysm within 12 hours),were underwent emergent PTCA(coronary stem performed in some patients),including 18 cases which were treated combined with Lipo-PGE_1 in the mean time.And the clinical efficacy and the results of short-period follow-up were recorded.Results The in- farctive vasculars were re-open in 37 patients(23 cares were routinely placed translunrinal srents),and the successful rate was 94.9 %.Those who also used Lipo-PGE_1 were re-open in 17 patients.The successful rate was 94.4 %,their ST segments on EKG 30 minutes after operations reduced significantly than that of patients who did not use Lipo- PGE_1,their cardial functions were also improved significantly 24 hours after operations and no side effects on blood pressure and heart rate were observed.Conclusion The emergent PTCA combined with Lipo-PGE_1 for acute my- ocardial infarction can protect the cardial function and show a better early therapy effect.

OBJECTIVESecond mitochondria-derived activator of caspase (Smac) is a recently identified, novel pro-apoptotic molecule, which is released from mitochondria into the cytosol during apoptosis. Smac promotes activation of caspases by neutralizing members of the inhibitor of apoptosis proteins (IAPs) family, such as X-linked inhibitor of apoptosis protein (XIAP). The objective of the study was to examine the pro-apoptotic effect of human Smac gene on Burkitt's lymphoma Raji cells.

METHODSThe full length cDNA of human Smac gene was amplified by reverse transcription-PCR from total RNA of HEK-293 cells. The PCR product was ligated with linearized vector pGEM-T-easy supplied in the TA cloning kit and sequenced. The correct cDNA of full length Smac was subcloned into eukaryocytic expression vector pcDNA3.1/myc-his and transfected into human Burkitt's lymphoma cell line Raji by lipofectamine-mediated transfection. The expression of full length Smac was determined by Western blot. Morphological observation was done with the laser scanning confocal microscope by double staining the Raji cells with Hoechest 33,258 and propidium iodide. Flow cytometry was used to evaluate apoptosis. Relative caspase-3 activity was determined by colorimetric assay.

RESULTSRecombinant eukaryocytic expression vector pcDNA3.1/Smac, which contained full length Smac, was successfully constructed. After pcDNA 3.1/Smac was transfected into human Burkitt's lymphoma Raji cell line for 24 hours, Raji cells showed apparent apoptosis with a percentage of (43.7 +/- 2.5)%, which was higher than that of non-transfected group and free vector-transfected group (P < 0.05). Compared with non-transfected group (0.136 +/- 0.036) and free vector-transfected group (0.138 +/- 0.026), the relative caspase-3 activity of Raji cells transfected by pcDNA3.1/Smac (0.936 +/- 0.041) was significantly enhanced (P < 0.05).

CONCLUSIONTransfection and expression of human Smac gene could significantly induce apoptosis of human Burkitt's lymphoma Raji cells. The mechanism is associated with the increase of caspase-3 activity.

Apoptosis ; genetics ; Blotting, Western ; Burkitt Lymphoma ; genetics ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; DNA, Complementary ; Flow Cytometry ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Mitochondrial Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods