AIM:To study the protection of Glycyl-L-Glutamine(Gly-Gln) against myocardial ischemia/reperfusion(I/R) injury in the isolated rat heart.METHODS:A model of myocardial ischemia-reperfusion injury was established with a Langendorff apparatus.Thirty male SD rats were randomly divided into four groups:control group,Gly-Gln group,I/R group and I/R+Gly-Gln group.Both I/R and I/R+Gly-Gln group were pre-perfused for 30 min,followed by 20 min ischemia and 40 min reperfusion.During reperfusion I/R+Gly-Gln group was perfused with Gly-Gln perfusate.Control group was kept perfused for 90 min.Gly-Gln group Gly-Gln perfusate was also kept perfused for 90 min.The left ventricular end-diastolic pressure(LVEDP),left ventricular developed pressure(LVDP),?dp/dtmax,heart rate(HR),monophasic action potentials(MAP) was measured during perfusion.The coronary effluent fluid was collected at different certain times.The activities of lactic dehydrogenase(LDH) and creatine kinase(CK) were determined.RESULTS:The isolated rat heart function decreased severely after 20 min ischemia and 40 min reperfusion(I/R):the LVEDP increased and the LVDP,?dp/dtmax decreased.But the LVEDP decreased and the LVDP,?dp/dtmax increased in I/R+Gly-Gln group compared with I/R group.Moreover,the activities of LDH and CK in the coronary effluent fluid decreased remarkably in I/R+Gly-Gln group compared with I/R group.CONCLUSION:Gly-Gln can play a protective role against myocardial I/R injury in isolated rat hearts via maintaining the left ventricular function and decreasing the release of LDH and CK.

Objective To develop a convenient, rapid and specific method using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for detection of facioscapulohumeral muscular dystrophy(FSHD). Methods Genomic DNA was extracted and digested by restricted endonuclease EcoR Ⅰ , followed by agarose electrophoresis. The DNA (< 38 kb) was retrieved from agarose electrophoretic gels. The primers and probe were designed in D4ZA gene in chromosome 4. One hundred and fifteen subjects were examined by FQ-PCR using the retrieved DNA (<38 kb) as a template and the result was analyzed by fluorescent curve comparing with positive control. Results The results by FQ-PCR showed that 13 cases were positive in 16 FSHD cases whose EcoR Ⅰ fragment sizes were known, 75 cases were negative in 78 cases of normal controls, 15 cases were positive in 16 FSHD cases diagnosed clinically whose EcoR Ⅰ fragment sizes were unknown, and 3 cases were positive in 5 cases of relatives of FSHD patients. Consistency was checked using Kappa index between the 2 gene diagnostic tests for FSHD (FQ-PCR test and the traditional Southern blotting test), and between the 2 diagnostic criterions (gene diagnosis by FQ-PCR and clinical diagnosis). The results were statistically significant (κ = 0. 765, P = 0. 002 ; κ = 0. 844, P = 0. 000). Conclusions A new genetic diagnostic method of FSHD by FQ-PCR was developed, which was more simplified and reliable compared to the time-consuming, radioactive Southern blotting. It could also detect the D4Z4 arrays in cases having deletion of p13E-11 as well as the interchromosomal exchange between 4q35 and 10q26. The new method of FQ-PCR for FSHD may be extended to utilize clinically in future.

Objective To transplant the umbilical cord mesenchymal stem cells(UCMSCs) derived from human umbilical cord into cisterna magna of hypoxic-ischemic encephalopathy(HIE) rat model,and to observe their survival,proliferation and differentiation in the rat brain.Methods UCMSCs were isolated from human umbilical cord of babies delivered after full-term normal cesarean section,and labeled by bromodeoxyuridine(BrdU).Pregnant rats were randomly divided into experimental group(n=6) and control group(n=1).HIE models were built by ligating both sides of the uterine arteries of full-pregnant rats(21 days) in experimental group rats for 15 minutes.The neonatal rats in experimental group were divided into stem cells group(n=24) and PBS group(n=19) at random.The labeled UCMSCs were injected into cisterna magna of the rats in stem cells group,while PBS was injected into the rats of PBS group.In 1,2,3 and 4 weeks after transplantation,the brain tissue section slides were immunohistochemically stained with antibodies against BrdU,Nestin,neuron specific enolase(NSE) and glial fibrillary acidic protein(GFAP),and thionin.Control group with normal delivery was tested as concurrent control.Results At 1 week after transplantation,BrdU,Nestin,NSE and GFAP positive cells were found in the hippocampal dentate gyrus of the rats in stem cells group rats.The number of BrdU-positive and Nestin-positive cells increased(Pa0.05).The NSE-positive and GFAP-positive cells gradually increased from 1-4 weeks post transplantation and comparisons between groups had statistical significance(Pa

Objective To analyze the local control rate and the desimetric patterns of local recurrence in nasopharyngeal carcinoma(NPC)patients having been treated with standardized conventional radiotherapy and to evaluate the delineation of rational target volume.Methods From Jan.2000 to Dec.2000,476 patients with untreated NPC were treated by standardized conventional radiotherapy alone at the Sun Yat-sen University Cancer Center.The radiation ports were designed on a X-ray simulator.The nasopharyngeal lesion demonstrated by CT scan and the subclinical spread regions adjacent to the nasopharynx were defined as the target volume.Kaplan- Meier method was used to calculate the cumulative local recurrence rate.For patients with locad recurrence,the primary and recurrent local tumor volumes(V_(nx),V_(recur))were delineated with three-dimensional treatment planning system(3DTPS),and the dataset of radiation ports and delivered prescription dose to the 3DTPS were transferred according to the first treatment.The dose of radiation received by V_(recur)was calculated and analyzed with dose- volume histogram(DVH).Local recurrence was classified as:1.“in-port”with 95% or mere of the recurrence volume((recur)_V_(95))was within the 95% isedase;2.“marginal”with 20% to95% of _(recur)V_(95)within the 95% isedese; 3.“outside”with only less than 20% of _(recur)_V_(95)within the 95% isodose curve.Results With the median follow- up of 42.5 months(range 8～54 months),52 patients developed local recurrence.The 1-,2-,3 and 4-year cumulative local failure rate was 0.6%,3.9%,8.7% and 11.5%,respectively.Among the 42 local recurrent patients who could be analyzed by 3DTPS,52% were in-port,40% were marginal and 7% were outside.For most of the marginal recurrence and all the outside recurrence patients,the main reason of recurrence were related to the unreasonable design of the radiation port and inaccuracy in the interpretation image findings.Conclusions The outcome of better local control rate and the dosimetric pattern of local recurrence show that the target volume is reasonable for NPC in Sun Yat-sen University Cancer Center.Enhancing the capability of correct interpretation of images,accurate design of the radiation pouts and making most useful molecular or functional imaging techniques to escalate the local radiation dose are promising ways to improve the local control further and better.

Objective To observe the safety and efficacy of oncolytic adenovirus (H101) implantation under EUS guidance combined with gemcitabine in patients of non-operative pancreatic cancer.Methods From May 2007 to December 2007,6 patients with non-operative pancreatic cancer were enrolled in the study.H101 were implanted into 3 sites of the tumor under EUS guidance.Gemcitabine Was siren systemicly on d2,d9 and d16 after implantation, and repeated 1 month later.Tumor size and perfusion were assessed by computed reconstruction and perfusion imaging.Changes of clinical indices,adverse effects and complications were also recorded.Results All patients completed the two periods of treatment as planned.Tumor size decreased in 5 cases(18.21%-38.65%),but without statistical difference (P=0.078).The area of liver metastasis Was found decreased in 1 ease.Perfusion imaging showed significant increase of mean transit time(P=0.049) and improvement in blood flow,blood volume and permeability surface at 2 weeks after the treatment. KPS increased in 2 patients and pain score decreased in 3 patients.Three patients died 2.5,2.5 and 3 months respectively after the procedure.while 3 other patients are still alive with the survival time of 3,5 and 10 months. Major adverse effects associated with H101 implantation were fever and flu-like symptoms.Mild acute pancreatitis occurred in 1 cage.Conclusion EUS guided oncolytic adenovirus implantation in advanced pancreatic cancer is feasible and safe.With the combination of gemcitabine,it is capable of shrinking the tumor size,destroying the angiogenesis of the tumor and improving the patients'living quality.

This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person.
Adolescent ; Bone Marrow ; pathology ; Bone Marrow Cells ; cytology ; metabolism ; Cell Culture Techniques ; Child ; Child, Preschool ; Female ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured ; Young Adult

OBJECTIVESTo study the role of hepatic sinusoid capillarization during the formation of portal hypertension in fibrotic rats induced by dimethylnitrosamine (DMN).

METHODSHepatic fibrotic rats were induced by administration of DMN intraperitoneally three times a week for 4 weeks. The rats were harvested on day 2 and weeks 1, 2, 3, 4, 5, 6, 8, 12 and 24. The formation of liver fibrosis and hepatic sinusoid capillarization were observed by morphologic methods. Pressure of portal vein (Ppv) was directed measured with intubation tube method by mesentry anterior vein.

RESULTSThe Ppv was getting higher and higher with the administration of DMN. After four weeks, the Ppv was higher than that of control [(1.10+/-0.18)kPa vs (0.52+/-0.04)kPa, t=6.41, P<0.01]. The dynamic change of hepatic sinusoid capillarization was in accordance with that of Ppv, which normalized gradually after the DMN was stopped. Significant positive correlation existed between the dynamic change of Ppv and the expression of vWF, laminin and alpha-SMA in sinus (r=0.833, P<0.01; r=0.953, P<0.01; r=0.919, P<0.01).

CONCLUSIONHepatic sinusoid capillarization is the vital cause for portal hypertension in fibrotic rats induced by DMN.

Animals ; Capillaries ; pathology ; Dimethylnitrosamine ; Hypertension, Portal ; chemically induced ; etiology ; pathology ; Liver ; blood supply ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; complications ; pathology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo assess the association between CSPG2 and HSPG2 gene polymorphisms and intracranial aneurysm (IA) in ethnic Han Chinese population.

METHODSA case-control study was carried out. A total of 537 IA patients and 1071 normal controls with matched age and gender were recruited. Peripheral blood samples were obtained from all subjects. Following extraction, target DNA was amplified with PCR and genotyped with a SNaPshot method. The association between 2 tag SNPs (rs251124 and rs3767137) of CSPG2 and HSPG2 genes and IA was assessed.

RESULTSThe genotype frequencies of rs251124 and rs3767137 were both in Hardy-Weinberg equilibrium. No significant difference has been found in the frequencies of rs251124 of CSPG2 between the two groups. Similarly, the frequency of rs3767137 (HSPG2) did not differ between the IA and control groups (P=0.22), albeit with an OR value of greater than 1 (OR=1.12, 95%CI=0.92-1.37). There were no significant difference in genotypic frequencies of the two SNPs between the two groups (P=0.46, 0.53).

CONCLUSIONNo association has been found between polymorphisms of rs251124 and rs3767137 loci of CSPG2 and HSPG2 genes and IA in the selected population.

Adult ; Aged ; China ; ethnology ; Female ; Heparan Sulfate Proteoglycans ; genetics ; Humans ; Intracranial Aneurysm ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Versicans ; genetics

OBJECTIVETo evaluate the effects of acupuncture on post-stroke spastic paralysis.

METHODSA systematic evaluation including all the relavant randomized controlled trials (RCTs) or quasi-RCTs of acupuncture and moxibustion for treatment of post-stroke spastic paralysis were carried out according to the method recommended by the Cochrane Collaboration.

RESULTSNine hundred and seventy-eight patients being included in fourteen papers met the enrolled criteria. However, their methodological quality was relatively poor. Meta-analysis of nine trials indicated that there was no significant difference between the treatment groups and the control groups in Ashworth scores, Carr-Shepherd scores, nerve defect scores and hip adductor tension scores. Whereas the Fugel-Meyer scores in one trial and the Barthel scores in three trials were better in the treatment groups than those of the control group.

CONCLUSIONA reliable conclusion can not be drawn from the present data because of the defects in methodological quality and insufficient numbers of trials, especially lack the long-term terminal outcomes, although it appears a tedency that acupuncture can improve the conditions of post-stroke spastic paralysis. Therefore, it is necessary to perform more multi-central RCTs of high quality in future.

Acupuncture Therapy ; Cerebral Palsy ; etiology ; therapy ; Humans ; Randomized Controlled Trials as Topic ; Stroke ; complications ; Treatment Outcome

Rabies virus ; physiology