BACKGROUND:Separating umbilical cord blood stem cells using tubes has low efficiency,and microbial contamination easily occurs during this process,therefore,safety cannot be ensured in clinical application.lt is urgent to find a method for separating umbilical cord blood stem cells to treat femoral head necrosis.OBJECTIVE:To establish a high efficient,safe,and clinically valuable method to separate umbilical cord blood stem cells.DESIGN,TIME AND SETTING:A self-control experiment was performed at the First Department of Surgery,Zhengzhou Second People's Hospital,Institute of Blood Constituent Application,Henan Red Cross Blood Centre between February 2006 and August 2007.PARTICIPANTS:Eight male patients with femoral head necrosis,averaging 40.6 years of age,were included in this study.Of these patients,4 had the history of hormone application.An average of 90 mL umbilical cord blood was harvested from each healthy normal full term neonate from Maternal and Children Health Care Hospital of Zhengzhou City.The quadruple bags used for separating umbilical cord blood stem cells consisted of 1 main bag,1 empty bag,and 2 physiological saline bags,provided by Shandong Weigao Holding,China.METHODS:Within 6 hours after collection,umbilical cord blood was centrifuged in the empty bag of quadruple bag,which was connected with an aseptic filling machine.After centrifugation,partial blood plasma was discarded,and the remaining erythrocytes were thoroughly mixed by adding hetastarch.Five minutes later,the mixture was diluted with physical saline at 1:1.Umbilical cord blood was slowly added into the main bag (at 1:1),in which,human lymphocyte separating medium was pre-added.After cantrifugation,the upper layer of solution,i.e.,monocyte-rich solution,was transferred into another empty bag.Within 24hours of preservation,after suspension with umbilical blood plasma,umbilical cord monocytes were transfused into patients with femoral head necrosis via superficial vein on the hand back,monocytes≥1×108/portion,2 portions once.There were three treatment courses,each involving three transfusion sessions,one session every 4 days,and a 2-3-month interval between two treatment courses.MAIN OUTCOME MEASURES:Cell recovery rate and cell viability of umbilical cord blood monocytes and improvements in clinical symptoms.RESULTS:The separation of quadruple bags could obtain umbilical cord blood monocytes with high recovery rate.Furthermore,microbial contamination hardly occurred in the process of separation.Hip joint pain relieved or disappeared to different extents in all 8 patients,with an effective rate of 100%.Abduction and internal rotation of hip joint,ambulation distance,and gait were markedly improved.At 6 months after cell transplantation,5 patients presented with changed bone density in femoral head necrosis regions,2 showed normal femoral head morphology,and the remaining 1 exhibited no obvious changes.Joint effusion was reduced or disappeared in 12 hips.Magnetic resonance images showed that femoral head morphology had been improved in various degrees in 9 hips,but no changes in 3 hips.No complications,fever,or allergies occurred during and after cell transplantation.CONCLUSION:The method of separating stem cells from umbilical cord blood in junction with aseptic interface technology is highly effective,safe,and clinically valuable.Multiple intravenous transfusions of umbilical cord blood stem cells provide a novel approach for systemic treatment of femoral head necrosis.

We have found the parallel relationship between the live cell number of several tumor cell lines and the formation of MTT formazan with MTT colorimetric assay. The MTT colorimetric assay was compared with ~3H-Tdr incorporation assay for the proliferation of mouse spleen cells induced by mitogens or the activity of rat/mouse IL2,the results suggested that the ~3H-Tdr incorporation assay can be replaced by MTT colorimetric assay. MTT colorimetric assay have the advantages of simplicity, rapidity,save any radioisotope and no specific equivepment, etc, so under some conditions it would be a useful method for measuring cell proliferation or cytotoxicity in a laboratory.

Objective:To put forward some suggestions for the national health development in the filed of health expenditure indicators .Methods:Two health expenditure indicators and four health outcomes indicators were chosen from WB and WHO official websites , and researches on the relationship between health expenditure and outcomes were conducted .We put forward proposed value of the health cost indicator .Results: With different levels of GDP per capita , the scatter diagram of health expenditure and GDP per capita had different rules .When health outcomes were taken into consideration , the rules were the same .When GDP per capita was at different level , better health outcomes were not related to higher health expenditure .Input-output ratio should be taken into consideration fully . Suggestions:The study should be dynamic as GDP develops .More factors can be added in the research , if the data of the factors is available .In this study , the GDP per capita was divided into 4 sections .The division standard of the GDP per capita can be more delicate .What is more important for the development of health expenditure is not contin-uing input , but better input-output ratio because of diminishing of marginal returns .One set of standards cannot be a-dapted to all countries and districts .We should give full consideration to the improvement of people's health instead of increasing input because the ultimate goal is the former .

Objective To discuss the methods of pre-hospital emergency care and transportation for patients with sudden chest pain. Methods Retrospective analysis was used in this study. The mortality was compared between 36 patients with sudden and deadly chest pain aided by pre-hospital emergency department and 21 patients with similar symptom aided by themselves. Results In indicators containing vital signs, severity of chest pain, mortality, the pre-hospital emergency care group was better than self-rescue group. Conclusions The clinical symptoms of patients with sudden and deadly chest pain are complex. However, professional pre-hospital emergency care can effectively reduce the mortality by closely monitoring, safely and effectively referred.

To gain a better understanding of the genetic diversity and evolution of PRRSV in the Ningxia Hui Nationality Autonomous Region (Ningxia) of China, the nsp2 genes from a series of PRRSV strains collected from the region in 2007 were partially sequenced. These sequences were then analyzed along with the classical strain (ch-la) and two other epidemic strains SD (3) and SD2006. Comparison of the nucleotide sequence with ch-la indicated that nsp2 genes of seventeen Ningxia isolates (NX strain) have deletions of 87 nucleotides. Sequence analysis indicated that homology between the Ningxia strain and ch-la was 60.3%-79.9% in the nucleotide sequence, and homology between the NX strains and SD strains was 80.3%-98.8% in the nucleotide sequence. The nsp2 genes of the seventeen isolates had 74.9%-100% nucleotide sequence identities with each other. This study was undertaken to assess the regional variation of prevalent PRRSV and to establish a sequence database for PRRSV molecular epidemiological studies.

OBJECTIVETo explore therapeutic effect and action mechanism of regulating spleen-stomach needling on diabetic nephropathy (DN).

METHODSUsing multi-centric, randomized, controlled and blind principles, 144 cases of DN were divided into an observation group and a control group according to random digital tab, 72 cases in each one. Based on regular treatment of diabetes, the regulating spleen-stomach needling was applied at Zhongwan (CV 12), Quchi (LI 11), Hegu (LI 4) and Xuehai (SP 10), etc. in the observation group while Shenshu (BL 23), Taixi (KI 3), Sanyinjiao (SP 6), Yanglingquan (GB 34), etc. were selected in the control group by reference of Acupuncture and moxibustion. The treatment was given twice a day, six days as a treatment session with interval of one day between sessions. Totally six weeks were required. Changes of clinical symptoms and signs, fast blood glucose (FBG), urinary albumin excretion rate (UAER), beta2-microglobulin (beta2-MG), monocyte chemotactic protein-1 (MCP-1), lymphocyte membrane cholesterol, propanediol (MDA), PCO, 8-hydroxydeoxy guanosine (8-OHdG), superoxide dismutase (SOD), CD3+, CD4+, CD8+, and CD4+/CD8+ were observed before and after treatment in two groups.

RESULTSAs for improving clinical symptoms and signs, total effective rate was 84.29% (59/70) in the observation group and 55.56% (40/72) in the control group, which had statistical difference between two groups (P<0.01). As for regulating glycometabolism [(6.25 +/- 0.32) mmol/L vs (8.09 +/- 0.63) mmol/L], reducing UAER [(154.43 +/- 55.14) mg/24h vs (268.91 +/- 77.65) mg/24h], restraining over-expression of MCP-1 [(137.59 +/- 36.15) pg/mL vs (166.89 +/- 42.82) pg/mL], regulating level of oxidative stress, prohibiting oxidation of protein and adjusting quantity and activity of T lymphocyte subgroup, the observation group was superior to the control group (P< 0.05, P<0.01).

CONCLUSIONThe regulating spleen-stomach needling is an effective method for treatment of DN, which cold improve glycometabolism disturbance-induced progressive kidney injury, recover glomerular filtration, reduce urinary albumin excretion rate, restrain overexpression of MCP-1, adjust level of oxidative stress, prohibit oxidation of protein, increase protectiveness of membrane, adjust quantity and activity abnormity of T lymphocyte subgroup, leading to repairing lymphocyte damage and improving immune expression to delay kidney damage.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Diabetic Nephropathies ; immunology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology

Objective:To investigate the role of extracellular signal-regulated kinase 1/2(ERK1/2)signal pathway in the pathogenesis of stress-induced gastric ulcer.Methods:Animal model of stress-induced gastric ulcer was established in rats with water-immersion restraint(WIR)stress.The mucosal activation of ERK1/2 was observed before and 5,15 and 30 min,and 1, 2 and 3.5 h after WIR stress.Some animals were also treated with an intravenous injection of PD98059(1 mg/kg),a specific ERK1/2 inhibitor,1 h prior to WIR stress.Expression of total ERK1/2 and caspase-3 were detected by Western blot analysis; ERK1/2 activity was measured by kinase activity assay using myelin basic protein as a specific substrate.DNA-binding activities of the transcription factors activator protein-1(AP-1)and nuclear factor-?B(NF-?B)were determined by electrophoretic mobility shift assays(EMSA).Mucosal TNF-?and IL-1?mRNA expression was analyzed by Northern blot analysis.The degrees of the gastric mucosal lesions were expressed as ulcer index(UI)and pathological evaluation.Apoptosis in the gastric mucosa was examined by an in situ TdT-mediated dUTP nick-end labeling(TUNEL)method.Results:Activated ERK1/2 was very weakly expressed in the gastric mucosa of normal rats.ERK1/2 was rapidly activated in the gastric mucosa of rats 15 min after WIR stress and the activity reached the maximal after 3.5 h.Pretreatment with PD98059 significantly inhibited ERK1/2 activation,decreased AP-1 and NF-?B activities and TNF-?and IL-1?mRNA expression,and obviously relieved gastric mucosal lesions,accompanied by caspase-3 activation and increased apoptosis.Conclusion:The present results indicate that ERK1/2 activation plays an important role in the development of stress-induced gastric ulcer.

Objective To investigate the effect of microRNA(miR)-21 on proliferation and differentiation of murine pulmonary fibroblasts. Methods C57BL/6 mice of SPF grade (n=24) were randomly divided into Sham group and Pulmo?nary fibrosis model group with 12 mice in each group. Pulmonary fibrosis model was established by trans-tracheal jet ventila?tion of bleomycin into mice. The transcription levels of miR-21 were examined by quantitative real-time PCR in various pul?monary fibrosis tissues. Primary fibroblast were isolated and digested by Trypsin then inoculated into 6 well plate to reach confluence of 30%-50%. PBS (2.5μL), negative control stock solution and miR-21 mimic stock solution (20μmol/L) were added into Opti-MEM (50μL) as control group, blank group and miR-21 mimic group respectively.The cell viability was as?sessed by CCK-8. Expressions of ADAMTS-1 and TGF-β1 in the pulmonary fibroblasts were tested using Western blot. Re?sults The expression of miR-21 was significantly increased in lungs of mice in pulmonary fibrosis model group than that in sham group. Expression of miR-21 was higher in miR-21 mimic group than that in control group and blank group. Expres?sion of miR-21 was significantly higher with better cell viability in miR-21 mimic group than that in control group and blank group. The expression of ADAMTS-1 was significantly decreased in miR-21mimic group, while the expression of TGF-β1, a target gene of miR-21, was significantly increased in miR-21 mimic group compared with the other two groups. There is no significant different in expressions of ADAMTS-1 and TGF-β1 between control group and blank group. Conclusion Over?expression of miR-21 in pulmonary fibroblasts disrupts TGF-β1 signaling pathway by reducing expression of ADAMTS-1, which promotes the proliferation and differentiation of pulmonary fibroblast.

Objective To evaluate the effects of intra-aortic balloon counterpulsation (IABP) on mortality of patients with acute myocardial infarction (AMI) treated with percutaneous coronary intervention (PCI).Methods Randomized controlled trials (RCTs) of IABP compared with non-IABP control in AMI patients, from January 1970 to May 2015, were searched from MEDLINE, Embase and Web of Science.The data were analyzed with software RevMan 5.0.Results Five RCTs involving 1 450 AMI patients, including 722 treated with IABP (IABP group) and 728 without IABP (non-IABP group), were included for analysis.Compared with non-IABP group, IABP did not significantly decrease the hospital mortality or 30-day mortality (OR=0.92, 95%CI: 0.69-1.25,P=0.61).According to the timing of IABP before or after PCI, it was further divided into IABP-before-PCI subgroup and IABP-after-PCI subgroup.Compared with non-IABP group, the 30-day mortality was not decreased in IABP-before-PCI subgroup or in IABP-after-PCI subgroup (OR=0.64, 95%CI: 0.23-1.78,P=0.39;OR=1.25, 95%CI: 0.42-3.77,P=0.69, respectively).According to complicating with cardiogenic shock (CS) or not, patients were divided to AMI with CS subgroup and AMI with no-CS subgroup;the hospital or 30-day mortality were not significantly decreased in both subgroups (OR=0.96, 95%CI: 0.70-1.32,P=0.80;OR=0.68, 95%CI: 0.28-1.70,P=0.27, respectively).Conclusion IABP does not decrease the 30-day mortality of AMI patients treated with PCI.

Objective To explore the relationship of methylenetetrahydrofolate reductase (MTHFR)gene C677T,factor V(FV)gene G1691A and prothrombin(PT)gene G20210A polymorphisms to unexplained recurrent early spontaneous abortion(URESA).Methods One hundred and twelve patients with URESA and 100 women with at least 1 normal pregnancy and without any miscarriage were analyzed for MTHFR,FV and PT gene polymorphisms by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Results MTHFR gene T/T genotype and T allele frequencies were increased in URESA patients[38.4%(43/112)and 59.8%(134/224)]versus controls[18.0%(18/100)and 43%(43/100),P