Comet Assay ; DNA Damage ; DNA Repair ; In Situ Hybridization, Fluorescence ; methods

Objective To establish the model of P2 peptide-induced experimental autoimmune neuritis(EAN)in rats and explore the roles of Th_1/Th_2 type eytokines in EAN.Methods Lewis rats were grouped into EAN rats and control rats.The EAN rats were immunized by injection into both hind footpads of inoculums containing 100 ?g or 200 ?g of P2_(57-81)peptide and FCA while the control rats were immunized with FCA only.Clinical scores were compared at the maximum of disease.Supernatant productions of IFN- ?, IL-4 and IL-10 secreted by lymphocytes and obtained on day 14 after the immunization were examined. Histopathological assessment of sciatic nerves was made.Results Peak clinical scores of P2_(57-81)200 ?g (3.6?0.3)group were significantly higher than P2_(57-81)100 ?g group(2.2?0.6,P

Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250－700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.

BACKGROUND: The role of mechanical stress in the functional regulation of osteoblasts becomes an emphasis in osseous biomechanical researches recently. There are few reports on whether fluid shear stress can induce the expression of core-binding factor α1 (Cbfα1) or not and what is its rule.OBJECTIVE: To investigate the effects of fluid shear stress (FSS) on the gene expression of Cbfα1 in human osteosarcoma cells.DESIGN: A controlled observation experiment.SETTING: Department of Aerospace Biodynamics, Faculty of Aerospace Medicine, Fourth Military Medical University of Chinese PLA.MATERIALS: Subjects were human osteosarcoma cells (MG-63) METHODS: This study was carried out at the Department of Aerospace Biodynamics, Faculty of Aerospace Medicine, Fourth Military Medical University of Chinese PLA from November 2004 to April 2005. ①After cultured for 60 hours, MG-63 were treated with 0.5 Pa (0.5 Pa stress treated group ) or 1.5 Pa (1.5 Pa stress treated group ) FSS in a flow chamber for 15, 30, 60minutes, respectively. Cover glass was put in the Petri dish containing culture medium at the same time, serving as the control group of FSS treated group. ②The total RNA in cells was isolated. Reverse transcription polymerase chain reaction analysis was made to examine the gene expression of Cbfα1 mRNA. The ratio of Cbfα1 mRNA and GAPDH mRNA was calculated.MAIN OUTCOME MEASURES: The mRNA expressions of Cbfα1 at different FSS and different time.RESULTS: ① Compared with control group, Cbfα1 mRNA expression increased significantly at 30 and 60 minutes with the treatment of FSS.Within certain time range (15 to 60 minutes), Cbfα1 mRNA expression was strengthened with the elongation of time and the increase of stress level. ②The Cbfα1 mRNA expression at 30 and 60 minutes were significantly increased in 1.5 Pa stress treated group in comparison with 0.5 Pa stress treated group (P＜0.01).CONCLUSION: FSS can significantly increase the gene expression of Cbfα1 in human osteosarcoma cells.

Objective To synthesize 4 kinds of 111 In?TPP cations and evaluate their properties as tumor cationic radiotracers in vivo and in vitro. Methods DO3A?xy?TPP, DO3A?xy?mTPP, DO3A?xy?dmTPP and DO3A?xy?tmTPP were radiolabeled with 111 In;their lipid?water partition coefficients and in vivo and in vitro stability were evaluated. The binding affinities of 4 kinds of 111 In?radiotracers were determined in cell uptake and cell efflux assay using U87MG tumor cells. Biodistribution studies and γ imaging studies were performed using the athymic nude mice bearing U87MG human glioma xenografts to explore the biologi?cal properties of 4 kinds of 111 In?radiotracers. One?way analysis of variance was used. Results The labeling yields of 4 kinds of 111 In?radiotracers were all above 85%, and the radiochemical purity were all greater than 99% after purification. Binding assay in U87MG cells showed that 4 kinds of radiotracers had great binding affinity and cell retention ability, and 111In?DO3A?xy?mTPP had the best binding ratio (1?49%;F=177.8, P<0.05) . Gamma imaging and biodistribution results showed that the U87MG tumors could be clearly visualized by 111In?DO3A?xy?mTPP, 111In?DO3A?xy?dmTPP and 111In?DO3A?xy?tmTPP, and the liver uptake of the 3 tracers was lower than that of 111In?DO3A?xy?TPP. In particular, 111In?DO3A?xy?mTPP had the best tumor/liver ratio (0.13±0.05, 2 h postinjection;F=9.4, P<0?05). Conclusions The tumor?targeted ability of 111In?DO3A?xy?mTPP is better than those of 111In?DO3A?xy?dmTPP, 111In?DO3A?xy?tmTPP and 111In?DO3A?xy?TPP, suggesting that it has the potential to be a promising tumor cationic radiotracer.

Objective To compare proximal femoral nail(PFN)introduction by percutaneous K-wire through a small incision with conventional PFN introduction protocol in the treatment of intertrochanteric fractures. Methods From January 2004 to March 2005,51 patients with intertrochanteric fractures were randomly dis- tributed into a minimally invasive treatment group(group MI)and a conventional treatment group(group C).All the fractures were closely reduced.In group MI a K-wire was percutaneously inserted through the tip of the greater troehanter into the center of medullary canal of the pruximal femur before the PFN was inserted under the guidance of K-wire through a small incision made along the K-wire while in group C the PFN was introduced according to the conventional procedure.The operation time,intra-operative blood loss,length of incision,X-ray exposure,duration of in-patient stay and time of bone union in both groups were recorded and compared.Results The mean oper- ation time,mean intraoperative blood loss and mean length of incisions in group MI were 77.20 min,104.20 mL and 5.12 cm respectively and significantly lower than those in group C(P＜0.01).The X-ray exposure and the reduction time in group MI lasted longer than in group C(P＜0.01).The mean time of bone union and in-patient stay in both groups were nearly equal(P＞0.05).At the latest tollow-up,all the fractures united in both groups without nonuuion or delayed union.Conclusion Compared with the conventional protocol,introduction of PFN by a pereutaneuus K-wire inserted into the central medullary canal of the proximal femur is much more minimally in- vasive and effective.

OBJECTIVETo explore the clinical efficacy of the arthroscopic Mender II stapler for the treatment of patients with meniscus anterior horn injury needing meniscal suture repair.

METHODSAmong 47 patients with meniscus anterior horn injury, 29 patients were male and 18 patients were female, ranging in age from 12 to 31 years old, with a mean age of (20.53± 4.12) years old. The duration of disease ranged from 3 to 35 days, and the average duration was (12.43±5.74) days. The Mender II stapler was used to carry out arthroscopic suture from outside to inside. The Lysholm knee scoring system was used to evaluate and analyze preoperative and postoperative symptoms, such as pain, limping embolism and so on.

RESULTSForty-six patients were followed up, and the duration ranged from 12 to 48 months, with a mean of (20.53±4.12) years. The incision healed at the first stage without important vessels and nerves injuries. The symptoms of the locked knee joint disappeared, and symptoms such as pain, limp, swelling and limitation of activity improved. The Lysholm score increased from preoperative 52.33±7.31 to postoperative 86.74±6.92.

CONCLUSIONUsing Mender II stapler to treat patients with meniscus anterior horn injury who were treated with arthroscopic suture from outside to inside is effective to improve symptoms, and to obtain good short-term results.

Adolescent ; Adult ; Arthroscopy ; methods ; Child ; Female ; Humans ; Male ; Menisci, Tibial ; surgery ; Suture Techniques ; Tibial Meniscus Injuries

Objective To observe the protective effect of hydroxysafflor yellow A(HSYA) on anoxia/reoxygenation (A/R) injury of neonatal primary cardiomyocytes, and its relationship with phosphoinositide 3-kinase/protein ki-nase B/glycogen synthase kinase 3β(PI3K/Akt/GSK3β) signaling pathway. Methods Primary cardiomyocytes of neonatal rats were isolated from the rats and incubated for 48 hours. The cells were adhered to each other and then divided into five groups:control group (Con group), anoxia/reoxygenation group (A/R group),HSYA treatment group(A/R+H group),PI3K inhibitor (LY294002)treatment group(A/R+L group)and HSYA+LY294002 treat-ment group (A/R+H+L group),then to collect the supernatant fluid of each group to measure LDH.The flow cy-tometry was used to measure the apoptotic cells. The protein levels of Bcl-2,Bax,Akt,p-Akt (Ser473),GSK3β, p-GSK3β (Ser9) were evalated by Western blot. Results A/R increased LDH release,the apoptosis rate (P<0.001),and the expression of pro-apoptotic protein Bax (P <0.001) with the decrease of anti-apoptotic protein Bcl-2,p-Akt(Ser473), p-GSK3β(Ser9)(P<0.001) as compared with the control group. HSYA treatment de-creased LDH release,the apoptosis rate (P<0.001),and the expression of Bax (P<0.001) and increase the ex-pression of Bcl-2,p-Akt(Ser473),p-GSK3β(Ser9)(P<0.001). Compared with the A/R+H group,the expres-sion of Bax was increased (P<0.001),while the expression of Bcl-2, p-Akt(Ser473), p-GSK3β(Ser9)was de-creased (P<0.001) in the A/R+H+L group. Conclusions HSYA protects rats'cardiomyocytes from anoxia/reoxy-genation injury by regulating PI3K/Akt/GSK3β signaling pathway.

Acinetobacter Infections ; etiology ; Acinetobacter baumannii ; drug effects ; isolation & purification ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Intensive Care Units, Pediatric ; Male ; Risk Factors

Objective To evaluate the effects of regular follow-up and systematic management in premature infants on early discovering of cerebral palsy and intervention efficacy in early phase. Methods Fifty premature infants with cerebral palsy recruited from early children development outpatient service, were classified as treatment group; 40 premature infants with cerebral palsy recruited from general outpatient service, were classified as control group. The infants of treatment group received systematic follow-up and intervention. while the infants of control group received no systematic management. Age and therapeutic efficacy were compared between the two groups. ResultsThe average age in the treatment group ( [4. 0 ± 1.2] months) was significantly younger than that of the control group ( [7.0 ± 1.4] months) ( P ＜ 0. 05 ). The mental development index ( MDI )and physical development index (PDI) in the treatment group was 91.8 ± 10. 2 and 90. 2 ± 11.2,respectively,which were significantly higher than that of the control group(80. 2 ± 11.7 for MDI and 79. 3 ± 10. 2 for PDI)( P ＜ 0. 05 ) . Conclusion Systematic management could help discover cerebral palsy in premature infants in time and could improve physical and mental development of these infants.