Objective To observe the protective function of murine compact bone derived-mesenchymal stem cells(MSCs) on bleomycin-induced lung injury.Methods MSCs were isolated from the mouse compact bone and then cultured.The immunophenotype and differentiation potential of MSCs were identified by flow cytometry and inductive culture.56 female C57BL/6 mice were assigned into eight groups(7 each): normal group,busulfan intraperitoneal administration group(BUS),bleomycin-induced lung injury group(BLM),bleomycin-induced lung injury and busulfan intraperitoneal administration group(BLM+BUS),BLM challenge and MSCs treatment group(BLM+MSCs),BLM challenge and busulfan and MSCs administration group(BLM+BUS+MSCs).According to infused cell number,BLM+MSCs and BLM+BUS+MSCs group were again respectively divided to two subgroups: BLM+MSCs 1 group(5?105 MSCs) and BLM+MSCs 2 group(2.5?105 MSCs),and BLM+ BUS+MSCs 1 group(5?105 MSCs) and BLM+BUS+MSCs 2 group(2.5?105 MSCs),respectively.The mice were sacrificed 14 days after MSCs infusion,and their lungs were removed for pathological analysis and quantification of hydroxyproline.Results Compact bone derived-MSCs were fibroblast-like cells,positive for Sca-1,CD44,CD29,and CD105,but expressed no CD34,CD45,CD11b and CD31.They could differentiate into adipocytes and osteoblasts in vitro.Compact bone derived-MSCs infused by tail vein were capable of protecting lung from injury.In BLM+MSCs group,2.5?105 MSCs infusion demonstrated more efficient protective activity than that of 5?105 MSCs;the lung injury was less severe in BLM+BUS+MSCs group than that in BLM +MSCs group.The content of hydroxyproline in lung tissue of normal,BUS,BLM,BLM+BUS,BLM+MSCs 1,BLM+MSCs 2,BLM+BUS+MSCs 1 and BLM+BUS+MSCs 2 group was 0.72?0.05,0.79?0.10,1.37?0.09,1.22?0.16,1.24?0.14,1.01?0.15,1.03?0.19 and 0.98?0.12?g/mg,respectively.The content of hydroxyproline increased in BLM and BLM+BUS group compared with that in normal group(P

AIM:To investigate the effect of tribu saponin from Tribulus terrestris(STT) on the lipoprotein lipase(LPL) and hepatic lipase(HL) activity in lipid metabolic disorder mice.METHODS:Mice being fed lipodiet were taken in STT through the experiment with Fenofibrate as positive control drug.Four weeks later,the levels of LPL in the plasma,the adipose tissue,liver tissue,muscular tissue and the levels HL activity in the plasma,liver tissue were estimated with LPL and HL Kit.RESULTS:HL activity in liver tissue in lipid metabolic disorder group was significantly lower than that in STT,Fenonbrate and normal group(P0.05).In STT group,the LPL activity in adipose was lower(P

AIM: To evaluate the relationships between maternal serum alpha-fetoprotein (MSAFP) levels and middle cerebral artery peak systolic velocity (MCA-PSV) in pregnancies with fetal anaemia and to compare the sensitivities of MSAFP and MCA-PSV for the predicting the risk of fetal anaemia. METHODS: Fifty-five measurements of MSAFP and MCA-PSV were carried out in 32 women at risk of fetal anaemia (4 cases of alloimmunisation, 11 cases of thalassemia, 10 cases of parvovirus infection and 7 cases of placental chorioangioma). The relationship between MSAFP and MCA-PSV was studied, and 19 fetal blood samples, in which MCA-PSV measurements were abnormal, were taken and the fetal heamoglobin were tested in order to evaluate the correlation of MSAFP and MCA-PSV. RESULTS: A correlation between MSAFP and MCA-PSV (n=55, r=0.57, P<0.01) was observed, in which 15 cases of fetal anaemia and 4 cases false positive (non-anaemia) were detected among the 19 fetal blood samples. The MSAFP levels of 4 false-positive cases were normal. The MSAFP levels in 15 fetal anaemia cases were higher than those in non-anaemia. The elevation of MSAFP level was 15-20 d earlier than that of MCA-PSV in the cases of alloimmunisation and thalassemia, and it was 10-12 d later in the cases of parvovirus infection and placental chorioangioma significantly (P<0.05). Both MSAFP (r=-0.87) and MCA-PSV (r=-0.67) were significantly correlated with fetal heamoglobin level. CONCLUSION: The MSAFP level is significantly correlated with both MCA-PSV measurements and fetal haemoglobin. The time and process of the elevations of MSAFP indicate that MSAFP is more sensitive than MCA-PSV to predict and monitor the pregnancies at the risk of fetal anaemia.

Objective To evaluate the efficacy of the parathyroidectomy (PTX) in the treatment of severe secondary hyperparathyroidism (SHPT) with Sagliker syndrome (SS). Methods A retrospective review was undertaken among 212 SS patients underwent PTX in our hospital and with more than 3 years' follow up. The definitions of the efficacy were based on the postoperative intact parathyroid hormone level (iPTH). Cure showed that the iPTH was ＜ 150 ng/L; marked effectiveness was 150-300 ng/L; effectiveness was 301-500 ng/L;ineffectiveness was ＞500 ng/L. The status was defined as persistent SHPT if iPTH was ＞ 150 ng/L after surgery. The status was considered as SHPT recurrence if iPTH was ＜ 100 ng/L in the first week after surgery, and gradually increased and ＞ 150 ng/L with the follow-up. Results ( 1) Ten patients were involved and the average dialysis time was 142 months [male/female: 4/6; age 30-54 (39. 3 ± 10. 4) years]. All patients had severe bone and joint pain, accompanied with progressive facial increases, chicken breast, kyphosis, hip bone deformities, and body height shortening. (2) Preoperative tests: the median of iPTH 2000(1800-2863) ng/L; serum calcium (2. 45 ±0. 21) mmol/L, phosphorus (2. 19 ±0. 51) mmol/L, alkaline phosphatase ( ALP) (1189. 8 ± 780. 0) IU/L. Two to four enlarged parathyroid glands were confirmed by ultrasound and 99Tcm-MIBI parathyroid scintigraphy. ( 3 ) Surgical procedures: local or general anesthesia for PTX. Supplement with calcium and calcitriol implemented low serum calcium after PTX. (4) Follow-up: symptoms, including bone pain, muscle weakness, skin itching, and insomnia, were significantly improved after surgery. Transient hoarseness occurred in 2 cases. The iPTHs of all patients were decreased significantly after surgery. The median of iPTH was 55.5 ( 10-967) ng/L at 1 month post PTX, and was significantly less than prior to PTX (P＜0. 001). Eight patients were cure , 1 marked effectiveness ,and 1 ineffectiveness. Two patients were persistent SHPT, and 1 died of heart failure in the 4th year after PTX. The development of bone deformities was stopped and malnutrition was improved in long-time follow up. The level of iPTH 135(28-390)ng/L(P＜0. 001 ) , serum calcium, phosphorus, and ALP showed normal in the third year. The SHPT recurrence was appeared in the 2nd and 3rd year in 2 out of 8 patients, respectively. Conclusions Total PTX can effectively treat SS by SHPT. It can improve prognosis for patients, such as bone pain disappearing, bone deformities stopping and malnutrition improving, etc. The level of iPTH may rise again in some patients in the future. Therefore, more attentions should be paid to monitoring.

BACKGROUND: There are several types of endogenons stem/progenitor cells in lung that develop from either endoderm or mesoderm precursors.To elucidate the characteristics of the intrapulmonary stem ceils wig help to understand their biological behavior in lung injury.OBJECTIVE: To isolate the mesenchymal stem cells (MSCs) from mouse lung tissues,and identify their morphology and growth characteristics,cell surface antigens,differentiation potential in vitro,stem cell properties,and to investigate the protective role in bleomycin challenged lung.DESIGN,TIME AND SETTING: The present randomized controlled in vivo animal experiment based on in vitro observation of cytology was performed at the Laboratory of Cell Biology,Institute of Basic Medical Science,the Academy of Military Medical Science between October 2005 and August 2007.MATERIALS: Male(3-4 weeks) and female C57BL/6 (6-8 weeks) mice were used.Twenty female C57BL/6 mice were randomly and evenly divided into a pulmonary MSCs (PMSCs)-treated group and a myelosuppression group.METHODS: The lungs from male C57BL/6 mice were digested with collagenase Ⅱ,followed by centrifugation over a Ficoll step.The interface fraction was collected and cultured by adherent method.When the monolayer of adherent cells reached 70%-80%confluence,the adherent cells were detected and expanded in culture medium.The mice in the PMSCs-treated and myelosuppression groups were intraperitoneally administered busulfan to inhibit the immigration of bone marrow stem cells.In addition,pulmonary fibrosis injury was induced in these mice with bleomycin.The PMSCs-treated group was intravenously administered 5×105 PMSCs.At the same time,the myelosuppression group received 100 μ L of phosphate buffered saline.After 14 days,the lungs were taken to prepare paraffin-embedded section.MAIN OUTCOME MEASURES: Cell morphology,immunophenotype,specific markers of the induced cells,expression of gene peroxisome proliferator activated receptor γ,osteopontin,osteocalcin,prosurfactant protein C(SP-C),Oct-4 and Nanog,histological alteration of lung tissue sections.RESULTS: MSCs were successfully isolated from mouse lung tissue.The pulmonary MSCs(PMSCs) were fibroblast-like cells,and expanded rapidly in vitro for up to 40 passages.The phenotype of the PMSCs was Scu- 1+ ,CD44+ ,CD29+ ,CD105+ ,CD54+ ,CD34-,CD45-,CD11b,c-kif,and CD31.They did not express alveolar epithelial cell specific markers,such as SP-C,aquaporin-5,and clara cell secretory protein.It was noticeable that stem cell markers octamer-binding transcription factor 4 (Oct-4) and Nanog were expressed continuously in culture expanded cells.They could differentiate into adipocytes,osteoblasts and alveolar epithelial cells in vitro.Colony-forming assay demonstrated that the colony forming efficiency of the PMSCs was nearly 3%.The cells from single colony were capable of differentiating to osteocytes,adipocytes and alveolar epithelial cells.Intravenous administration of PMSCs could alleviate pulmonary injury and fibrosis induced by bleomycin,despite the bone marrow was intact or not.CONCLUSION: The PMSCs are able to extensively propagate in vitro,can efficiently switch to both mesenchymal and alveolar epithelial lineages in vitro,generate adipocytes,osteocytes,and the alveolar epithelial cells,and reduce pulmonary injury and fibrosis in bleomycin challenged lung in vivo,strongly suggesting their promising applications in cellular therapy against the lung injury.

Objective To explore the common genetic abnormalities in childhood acute lymphoblastic leukemia(ALL) and their responses to early treatment response.Methods From December of 2010 to December of 2011,169 newly diagnosed ALL patients at the Department of Hematology,Beijing Children's Hospital Capital Medical University,were detected by karyotype analysis,reverse transcription polymerase chain reaction (RT-PCR) and fluorescent in situ hybridization (FISH),and the relationship between early treatment responses and genetic abnormalities was observed.Results Of the 169 cases,bone marrow cell specimens from 162 cases were successfully cultured,with the success rate reached to 95.9％,and 88 cases (52.1％) had chromosomal abnormalities.Fifty-five cases carried 8 types of fusion genes among the 153 patients who received RT-PCR examination,and the abnormal rate was 35.9％.Forty cases applied for the detection of mixed lineage leukemia (MLL) gene rearrangement by FISH,and 6 cases of them were positive.One hundred and five cases had genetic abnormalities and the detection rate reached to 62.1％ by using three combined methods.The genetic abnormalities were classified into 6 groups,they were t(12;21),t(1;19),t(9;22),MLL rearrangement,hyperdiploid and-6/6q-,-7/7q-respectively,and early therapy response in each group was compared,and statistically significant differences were found among 6 groups (x2 =22.954,19.432,14.045,P =0.001,0.001,0.016).Conclusions Conventional cytogenetics combined with RT-PCR and FISH can enhance the detection rate of genetic abnormalities in childhood ALL.Genetic abnormalities combined with early treatment response in ALL can better guide the clinical treatment and prognosis assessment.

AIM:To investigate the effect of NF-κB binding element deletion on transcriptional regulation of (NOX1).METHODS:pGL3-Basic vector inserted with the NOX1 proximal promoter, and the same vector inserted with the NOX1 proximal promoter in the absence of the positive NF-κB binding element, were constructed.After cloning, diges-tion and purification, NOX1 proximal promoter (≈1 415 bp) was inserted into the multicloning site of the pGL3-Basic vec-tor and then sequenced ( pGL3-NOX1-1415) .NF-κB binding elements in the NOX1 promoters were predicted by Alibaba 2.1 software.The positive element was deleted by overlapping PCR.The deletion mutant was inserted into the pGL3 vector in the same way (pGL3-NOX1-1327).The plasmids were transfected into A549 cells, and then the cells were stimulated with TNF-α.The luciferase activity was monitored on MD SpectraMax M5 enzyme-labeled instrument.RESULTS:The se-quences of pGL3-NOX1-1415 and pGL3-NOX1-1327 were identified to be correct.Compared with control group, the lucif-erase activity was significantly higher in the cells transfected with pGL3-NOX1-1327 (P<0.05), but it was significantly lower than that in the cells transfected with pGL3-NOX1-1415 (P<0.05).CONCLUSION: NF-κB plays an essential role in the transcriptional regulation of NOX1 in TNF-α-induced A549 cells.Activated NF-κB binds to specific elements in NOX1 promoter regions to control the transcription.

Objective To investigate the clinical values of multiple ultrasound soft markers in screening for fetal chromosomal abnormality during first-trimester.Methods Two thousand seven hundred and eighty-nine nulliparas in Department of Obstetrics,the First Affiliated Hospital of Jinan University during early pregnancy (11-13+6 gestational weeks) were selected for this study.Fetalnuchal translucency (NT),facial angle (FA),ductus venosus (DV),fetal heart rate (FHR),tricuspid reverse (TR),nasal bone (NB) and fetal structures were scanned and measured.Risk calculation software (Astraia) was used to calculate the chromosomal abnormal risk (cut-off line:＞1/300) based on ultrasound records.The chorionic villi or amniotic fluid of high risk patients was collected with informed consent for karyotype analysis (prenatal diagnosis).All patients were followed up until six months after delivery.Chi-square test or Fisher exact test was used to compare the difference.Results One hundred and seven cases were high-risk of trisomy 21 among which 96 cases accepted invasive prenatal diagnosis.Sixteen chromosomal anomaly and six trisomy 21 cases were diagnosed out the 96 fetuses.Among 2789 cases,four were high-risk of trisomy 21 according to ultrasound screening.Six cases were diagnosed as trisomy 21.The false positive rate of ultrasound screening was 3.6％(101/2783).There were 196 cases whose NT ≥2.5 mm,in which 66 cases were high risk of chromosomal abnormality,and 16 fetal chromosomal abnormalities were diagnosed after chorionic villus sampling.The invasive procedure rate was 2.3％ (66/2789).Totally,186 pregnant women were older than 35 years,among which 32 cases were high risk.There was no significantly difference on the of rate fetal chromosomal abnormality between the groups of age≥ 35 pregnant women and the general population (P=0.055).But 29.9％ (32/107) high risk cases were detected in the group of age≥35.Five of thirteen fetal malformations cases were associated with abnormal karyotype.Conclusions Multiple ultrasound soft markers screening during early pregnancy could increase the diagnosis rate of chromosomal abnormality and decrease the false positive rate,false negative rate and invasive procedure rate.Early ultrasound screening might be effective in not only identifying chromosomal abnormality,but also diagnosing severe structure deformity of fetus.

Objective To explore the occurence,clinical characteristics and treatment of Mycoplasma pneumonia complicated with embolism in children.Methods Twenty-three cases with Mycoplasma pneumonia complicated with embolization were retrospectively analyzed from January 1990 to December 2012.Results The ages of cases were from 4 years old to 13 years old,and fifteen cases were male,eight cases were female.Nineteen cases with single-shot embolism included four cases of lower limb venous thrombosis,one case of internal carotid artery thrombosis,nine cases of cerebral infarction,two cases of cardiac infarction,two cases of splenic infarction,one case of pulmonary infarction; and the other four cases were multiple embolism,two cases combined pulmonary embolism and lower limb deep vein thrombosis,one case combined cardiac embolism and pulmonary embolism,one case combined internal carotid artery and the brain embolism.In addition,eight cases had temporary anti-cardiolipin antibody IgM,two cases combined protein C decrease,one case merge protein S decrease,and one case was lack of AT-Ⅲ.At last,two children died,the rest all recovered well after thrombolysis and anticoagulation therapy.Conclusion Mycoplasma pneumoniae has hypercoagulative state and potentialized to thrombosis,especially for children with high risk factors of thrombosis.Early diagnosis and anticoagulation and thrombolysis treatment actively is the key to better prognosis.

Objective To isolate and identify leukemia stem cells from acute myeloid leukemia patients for further research. Methods By Ficoll density gradient centrifugation, mononuclear cells were firstly separated from bone marrow of patients. According to specific surface markers, CD+34 CD+123 of leukemic stem cells were sorted by flow cytometer. Their ability of self-renewal and differentiation were evaluated by colony formation and cobblestone forming ability. At the same time, the purity and cell morphology of CD+34 CD+123 cells was analysed. Results Comparared with total mononuclear cells, the proportion of the CD+34 CD+123 cells after sorting was 10.7 %, and these cells showed the ability of colony forming and cobblestone forming, and the purity proportion of CD+34 CD+123 cells was 62.1%. Conclusion The leukemia stem cells were isolated successfully and could be used in further study.