Objective To explore the value of laparoscopic diagnosis and treatment for Meckel’s diverticulum in children. Methods Laparoscopic exploration was performed in 11 children with suspected intestinal Meckel’s diverticulum. In other 2 children, the diverticulum was encountered during laparoscopic appendectomy, and then the umbilical incision was prolonged to take out the corresponding intestine for resection. Results No conversions to laparotomy were required in all the 13 children. The operation time lasted 30~90 min (mean, 75 min). The children were discharged at 5~6 postoperative days without complications. Conclusions Laparoscopic diagnosis and treatment for Meckel’s diverticulum in children is effective.

BACKGROUND: Previous researches indicate that ADp14ARF transfecting positive tumor cell line of p53 can inhibit the proliferation; in addition, the inhibitory effect is superior to transfection negative tumor cell line of p53. Whether simultaneous transfection of p14ARF and p53 genes can increase expression and accumulation of p53 and accelerate apoptosis of tumor cells needs further studies.OBJECTIVE: To construct double plasmid expression vector plRES-p14ARF-p53 by using gene engineering so as to observe the inhibitory effect on proliferation of osteogenic sarcoma cells.DESIGN: Randomized controlled observation.SETTING: Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Public Laboratory Platform, Immune Researching Room, Basic Medical College, Tongji Medical College, Huazhong University of Science and Technology from January 2005 to October 2006. Human osteogenic sarcoma MG-63 cells were provided by Cell Laboratory, Immune Researching Room, Tongji Medical College, Huazhong University of Science and Technology. plRES-p53 plasmid and plRES vector containing p53total-length gene order were provided by Wuhan Jingsai Biology Company.METHODS: Based on gene engineering, p14DNA (0.5 kb) was amplified from cultured L02 cells of normal human hepatic cells into plRES vector. Recombinant plasmid plRES-p14ARF-p53 was determined with polymerase chain reaction (PCR) and restriction enzyme and transfected into human osteogenic sarcoma MG-63 cells through mediation of liposome to screen positive clones. Otherwise, cells were divided into three groups, including blank control group (MG-63cells), blank vector control group (stably transfecting plRES-neo cells) and p14ARF-p53 group (stably transfecting plRES-p14ARF-p53 cells). ① DNA content and cycle of tumor cells were measured by using flow cytometry before and after transfection. ② Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect quantitative and semi-quantitative expression of p53 and p14ARF protein in tumor cells after stable transfection. ③Thiazole blue chromatometry and growth curve were used to observe proliferation.MAIN OUTCOME MEASURES: ① DNA content and cycle of osteogenic sarcoma cells; ② expressions of p53 and p14ARF protein in tumor cells; ③ proliferation.RESULTS: Double plasmid expression vector plRES-p14ARF-p53 was constructed successfully. ① DNA content and cycle of osteogenic sarcoma cells: Flow cytometry demonstrated that tumor cells mainly stayed in G1 phase after transfection. ② Protein expression: RT-PCR and Western blot indicated that p14ARF and p53 gene independently expressed in target cell mRNA and protein, respectively. ③ Cell growth: At 24, 48, 72 and 96 hours after MG-63 transfection, inhibitory rates of tumor cells were 33.43%, 69.37%, 66.19% and 75.26%, respectively, which was significant difference as compared with blank vector control group (P ＜ 0.01).CONCLUSION: Wild p53 and p14ARF can synergistically inhibit the proliferation and accelerate the apoptosis of osteogenic sarcoma cells.

Objective To investigate the predisposing alleles of HLA-DR genes in pemphigus. Methods Polymerase chain reaction specific sequence primers (PCR-SSP) method was applied to type HLA-DR subregion in the patients with pemphigus vulgaris (PV), pemphigus erythematosus (PE) and matched control subjects of Han nationality from Jiangsu and Anhui provinces. Results The results demonstrated that DR4, DRB1*14 (*1401, *1404,*1405)gene frequencies were significantly higher in both PV(Pc

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.
Biocompatible Materials ; Bone Marrow Cells/*cytology ; Bone Matrix/*cytology ; Cells, Cultured ; Chondrocytes/cytology ; Coculture Techniques ; Decalcification Technique ; *Osteogenesis ; Stem Cells/cytology ; Stromal Cells/cytology ; *Tissue Engineering

AIM:To investigate the influence of ovariectomy and estrogen replacemeot treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham(Ⅰ),ovariectomy(Ⅱ,OVX)and estrogen replacement treatment(Ⅲ,OVX+E2)group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters(18),cell receptors(9),intracellular transducers/effectors/modulator(7)and metabolism(6).Most of the genes(45)were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.

Objective To demonstrate the effect of comprehensive treatment under nasal endoscopy for epistaxis. Methods The bleeding sites of 92 patients were defined by nasal endoscopic examination. Epistaxis was cured by single pole or bipolar coagulation, combined with micro packing and systemic treatment. The bleeding sites and effect were studied retrospectively. Results The hemorrhagic foci were found in the following sites: 60.87%(56/92) in Little area, 13.04%(12/92) in the middle and back of nasal septum, 10.87% (10/92) in olfactory sulcus, 8.70% (8/92) in middle turbinate , 3.26% (3/92) at the top of inferior meatus, 2.17% (2/92) at the top of nasal cavity, 1.09% (1/92) at unknown part at the back of nasal cavity. Epistaxis was successfully controlled by once nasal endoscopic examation and hemostasis in 86 of 92 patients. While in 5 of 92 patients, epistaxis was cured by twice nasal endoscopic examation and hemostasis. Endoscopicligation of the sphenopalatineartery was performed in 1 patient with unknown posteriorepistaxis. In 92 patients,15 cases were given micro packing combined with systemic treatment. All the patients were cured and were followed up for 3 months without recurrence and the cure rate was 100.00%. Conclusions The major bleeding site is Little area. Single pole or bipolar coagulation, combined with micro packing and systemic treatment under nasal endoscope is effective for epistaxis and worth of clinic application extensively.

Objective To investigate the expression level of adenosine 2A receptors in peripheral blood lymphocytes and the correlation with disease progression of Parkinson's disease (PD).Methods In this retrospcctive study,out patients with PD(42 cases)and healthy controls(32 cases) were recruited at Beijing Hospital.The expression level of A2A receptors in peripheral blood lymphocytes was detected by flow cytometry.The concentration of free A2A receptors in plasma was detected by enzyme linked immunosorbent assay (ELISA).The expression of A2A receptors and plasma free A2A receptors in peripheral blood lymphocytes of the PD group and the control group was compared.Multivariate regression analysis and single factor correlation analysis were performed on sex,age,course of disease,duration of medication,drug type and dose,motor complications,and PD unified Parkinson's disease rating scale (UPDRS)score.Results A2A receptor expression in lymphocytes was significantly higher in the PD group than in the control group (8.96 ± 4.73)％ vs.(5.39±2.42)％ (t=4.210,P＜0.05).There was no significant difference in A2A receptor concentrations in plasma between the two groups (1.82 ± 1.91) μg/L vs.(1.15 ± 0.71) μg/L(t=1.078,P＞0.05).In the PD group,A2A receptor expression in lymphocytes in patients with motor complications was statistically lower than in patients without them (P＜ 0.05).Lymphocyte A2A receptor levels in the 5-9 years duration subgroup were significantly lower than those in the ＜5 years duration subgroup (Z=2.780,P＜0.01) and the≥10 years duration subgroup (Z=-2.149,P＜0.05).Conclusions The expression of A2A receptors in peripheral blood lymphocytes is correlated with PD.The expression of A2 A receptors in peripheral blood lymphocytes of patients with PD fluctuates with the occurrence of motor complications and the progression of disease.Further research is needed to establish A2A receptors as a biomarker for monitoring disease progression.

The operation methods, clinical classification, postoperative function exercise of gluteal muscles contracture were investigated. Clinically and retrospectively, treatment of 1280 patients with gluteal muscles contracture, being subjected to a "Z-shaped" release lengthening operation and efficiency exercise, was clearly standardized. All the cases were followed up from 3 months to 2 years with the effective rate being 100%, the cure rate being 98.5%, the recent complications being 5% , and the far complications being 0.2%. It was concluded that the clear diagnosis combined with standarized operation and efficiency functional exercise could greatly improve the therapeutic effects of gluteal muscles contracture.

The operation methods, clinical classification, postoperative function exercise of gluteal muscles contracture were investigated. Clinically and retrospectively, treatment of 1280 patients with gluteal muscles contracture, being subjected to a "Z-shaped" release lengthening operation and efficiency exercise, was clearly standardized. All the cases were followed up from 3 months to 2 years with the effective rate being 100%, the cure rate being 98.5%, the recent complications being 5% , and the far complications being 0.2%. It was concluded that the clear diagnosis combined with standarized operation and efficiency functional exercise could greatly improve the therapeutic effects of gluteal muscles contracture.

AIM: To study the molecular mechanism in modulation of expression of insulin receptor substrate-1 and-2 (IRS-1,-2) by estrogen and high concentration of insulin. METHODS: The 5′-regulatory regions of IRS-1 and IRS-2 gene were cloned into the pGL3 plasmid with luciferase reporter, and the clones were transfected into HeLa cells. The cells were incubated with estradiol (1 nmol/L) and high concentration of insulin (100 nmol/L). The relatively transcriptional activity of the 5′-regulatory regions of IRS-1 and IRS-2 gene was detected. RESULTS: It was found that the relatively transcriptional activity of the 5′-modulatory regions of IRS-2 reduced markedly after cells were incubated with 100 nmol/L insulin (P