BACKGROUND:The improvement of athletic achievements needs a super-limited exercise to gain,how to protect athletics health under the prerequisite of gaining excellent achievements?OBJECTIVE:To reduce the injury of athletic renal function following maximum exercise by using traditional Chinese medicinal methodDESIGN:A randomized and controlled trial,taking students majoring in sports education specialty as subjects.SETTING:Internal Medicinal Department of Traditional Chinese Medicine (TCM) Hospital of Qufu Hospital of Traditional Chinese Medicine and Sports Education College of Qufu Normal University.PARTICIPANTS :Two small natural classes were randomly drown up from 16 small natural classes of 2000 grade in Sports Education College of Qufu Normal University, with 20 students in each class, on April 20, 2004.METHODS:All 40 students were according to natural class divided as experimental class and control class.The urine specimens for test were collected 15-20 minutes after one time maximum loading training on April24,2004 and immediately after morning getting-up. The procedure was performed once more on May 24. During the process the students in experimental group had taken the capsule of kidney-and-spleen-strengthening Chinese drug (KSSD, containing mainly Rhizoma Dioscoreae, Radix Polygoni Multiflori, Herba Epimedii, Fructus Lycii, Gynostamma Pentaphyllum,Radix Acanthopanacis Senticosi,Radix Salviae Miltiorrhizae,Radix Bupleuri, etc., 0.3 g/pellet), 6 pellets, 3 times a day. Urinary glucoprotein,β2-microglobulin, albumin and globulin were detectedby radio-immunoassay.MAIN OUTCOME MEASURES:The changes of urinary proteins of the students before and after administration following maximum exercise and getting-up in next morning.RESULTS: All students involved were good for the trial, without any loss.① The contents of various urinary proteins for the subjects after the maximum exercise or before administration were all obviously increased,and the conditions of natural recovery of proteinuria on getting-up next morning were quiet different: the recovery of urinary albumin was the fattest, basically to the normal;recoveries of β2 micro-globulin and immune-globulin were obvious, but the recovery of glucoprotein was inconsiderable.② After the second trial,in comparing the experimental group with the control group it was found that KSSD can obviously influence the recovery of proteinuria following maximum exercise:The effect in 12 hours did not be shown because of too fast natural recovery of albuminuria. It could promote the recoveries of β2 micro-globulin and immune-globulin. The effect on recovery of urinary of glucoprotein was obvious,basically to the normal for experimental group and basically no change for control group,there was a significant differentiation between the two groups.CONCLUSION:The contents of urinary proteins were obviously increased following maximum loading training, and natural recoveries of the four kinds of proteins presented regulatory changes.KSSD can promote the recovery of proteinuria following maximum exercise to normal volume,suggesting that the medicine has a protecting role to renal function.

BACKGROUND: Abnormal hematopoietic microenvironment is an important factor causing dyshematopoiesis. However, no consensus has been reached on the sensitivity of hematopoietic stromal cells to irradiation.OBJECTIVE: To observe the changes of marrow stromal cells (MSCs) cycle and DNA content during the early stage of irradiation damage in mice, so as to further understand dyshematopoiesis due to radiation and provide scientific basis to avoid deleterious factors in hematopoietic environment.DESIGN: Completely randomized grouping and randomized controlled study based on the experimental animals.SETTING: Central laboratory of altitude military affairs medical department and altitude research institute of preventive medicine department, a military medical university of Chinese PLA.MATERIALS: This study was carried out at the Experimental Animal Center of Third Military Medical University between October 2002 and April 2003. A total of 60 healthy male Kunming mice were randomly divided into irradiation damage group and healthy control group, each having 30 mice.METHODS: The 30 mice in irradiation damage group were exposed to 60Co-γ of irradiation at a dose rate of 1.27 Gy/minutes within a distance of 4 m. Then the mice' marrow cells were harvested at day 3 and day 7 after irradiation, and were cultured in vitro for 14 days and 21 days for observation. Meanwhile the other 30 healthy mice unexposed to irradiation were considered as normal controls.MAIN OUTCOME MEASURES: Post-radiation number of MSCs colonies,cell cycle and DNA content.RESULTS: Although MSCs could grow and be adhered to walls after being exposed to irradiation of 5.0 Gy/s, the number of MSCs colonies was found significantly decreased compared to that of rnormal control group( P < 0.01 ).The colony number of the MSCs irradiated for 7 days obviously increased than that of MSCs irradiated for 3 days; however, MSCs recovered slowly and resulted in prolonged culture time, indicating the inhibited proliferation of MSCs due to irradiation damage. Results of flow cytometry showed that cells in G2+ M phase(2.60±0.41, 4.20±1.27) and DNA content (58.40±0.79,61.17 ± 1.35) in irradiation groups after 3-day and 7-day irradiation were obviously lower than those of normal control group(12.60 ±0. 75, 78.57±0. 83)(P <0.05-0.01).CONCLUSION: MSCs have relatively high sensitivity to irradiation damage and longer persisting period.

Objective:Our aim was to To clarify the roles of p16 and p15 proteins in the genesis of acute lymphoblastic leukemia(ALL).Methods:Twenty-three samples of ALL were studied by the method of indirect immunofluorescence.Flow cytometer was used to estimate the cellular fluorescent intensity to determine the levels of p16 and p15 proteins.Results:Negative expression for p16 protein was found in 10 of 23 samples,and 8 of 23 were p15 negative expression.Both kinds of proteins were abscent in 6 samples.2 of 3 cases of T-ALL were negative expression of p16,p15 protein.In non T-ALL,6 of 13 were negative expression for p16 protein,5 of 13 were p15 protein deficient.The expression rates of p16,p15 protein in high leukocyte group were lower than those of non-high leukocyte group(P<0.05).The expression rates of p16,p15 proteins in HR-ALL were lower than those of SR-ALL(P<0.05).Conclusion:The p16 and p15 proteins take part in the genesis of ALL.Negative expression of p16,p15 proteins might imply the poor clinical outcome.

AIM: To study the effect of mesenchymal stem cells (MSCs) infusion on hematopoietic recovery after peripheral blood stem cell transplantation in mice. METHODS: BALB/c mice conditioned by high dose chemotherapy/radiotherapy were infused with 10 6 peripheral blood mononuclear cells (PBMC) mobilized by granulocyte colony-stimulating factor (PBSCT group), 10 4 MSCs culture-expanded in vitro and 10 6 PBMC(experimental group 1), 10 6 MSCs and 10 6 PBMC(experimental gruop 2). Survival rate within 4 weeks, white blood cell count, bone marrow nucleated cells (BMNC), granulocyte-macrophage colony forming unit(GM-CFU) and fibroblast colony forming unit (F-CFU) were examined. RESULTS: Survival rate, BMNC, GM-CFU, F-CFU were significantly higher in experimental group 2 than that in PBSCT group ( P

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

Besides hematopoietic stem cells, bone marrow also contains another type of stem cells called mesenchymal stem cells (MSCs). With different induced conditions, MSCs have the ability to differentiate into a variety of nonhematopoietic tissue cells, including osteoblasts, chondroblast, adipocytes, myoblasts, astrocytes, and so on. MSCs can be readily obtained from bone marrow by their adhesion to plastic and expansion in culture. Also they can be genetically engineered by transduced target genes. MSCs may be the farget cells for both cell therapy and gene therapy for diseases derived from many different nonhematopoietic tissues.

Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases.MethodsBased on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (Dmd^Mu/+) mice were obtained after genotype identification. Male hemizygous Dmd^Mu/+(Dmd^Mu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting.An acute inflammation model was established in Dmd^Mu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry.Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (Dmd^Mu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of Dmd^Mu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, Dmd^Mu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old Dmd^Mu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old Dmd^Mu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old Dmd^Mu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old Dmd^Mu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups.Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients.