BACKGROUND: There are small amount of hematopoietic stem cells, which are easy to divide in vitro and show the difficulty in applying to transplantation.OBJECTIVE: This paper has focused on the role and means of the Wnt, Notch, Bmi_1, Shh, HOXB4 signaling molecule in the maintenance of hematopoietic stem cell self-renewal and regulation. METHODS: With the key words of "HSC, Wnt, Notch, Bmi_1, Shh, HOXB4" for the search, we searched PubMed database (2002-01/2008-12) in English. Literatures closely related to the hematopoietic stem cell self-renewal related signaling molecules were included. Repetitive research and Meta analysis were excluded.RESULTS AND CONCLUSION: The computer initially retrieved 216 documents, of which 30 documents for research. Hematopoietic stem cells are self-renewing, have strong differentiation and growth and regeneration capacities, can produce various types of blood cells ancestor cells, are widely used to treat blood diseases, but the hematopoietic stem cell differentiation in vitro demonstrated that the difficulties used in transplantation. How to make hematopoietic stem cells in vitro amplification and processing, while maintaining hematopoietic stem cell self-renewal characteristics is of a key issue. In recent years, different signaling pathways to enhance the ability of hematopoietic stem cell self-renewal signaling molecule have been a research hotspot. The article focused on the role and means of the Wnt, Notch, Bmi_1, Shh, HOXB4 in the maintenance of hematopoietic stem cell self-renewal and regulation and found that both the above-mentioned five signaling molecules can enhance hematopoietic stem cell self-renewal function. There are also a number of factors playing an important role in the maintenance of hematopoietic stem cell self-renewal process, such as endogenous factors, a series of transcription factors Oct-4, Ehox, Nanog, SCL, Runx1 and so on, to explore how their regulatory networks formed by the interaction control self-renewal of hematopoietic stem cells will become a key point in the research of self-renewal of hematopoietic stem cells.

BACKGROUND:Some studies show that basic fibroblast growth factor(bFGF)strongly expresses dudng the process of embryonic stem cells differentiation into hematopoietic stem cell.yolk sac blooding.and fetal liver hematopoiesis.OBJECTIVE:To study the regulation of bFGF on the blast-colony-forming cell(BL-CFC)by adding bFGF in the medium of embryoid body generation.METHODS:The third to fifth generations of the pdmaw mouse embryonic fibroblasts were recovered,and then incubated with the DMEM medium containing mitomvcin C for 2.5 hours in order to lose the proliferative capacity.Then cells were suspended into single cell by trypsinization and inoculated in the gelatin-coated bottle at the density of 10×104/cm2.After culturing for 24 hours,mouse embryonic stem cells(mESC)of D3 were recovered and placed on the feeder layer cells.According to the composition of medium in embryoid body generation.mESCs were divided into two groups:group A:standard medium+VEGF+SCF;group B:standard medium+VEGF+SCF+bFGF.Each group was cultured for 3 days and 6 days respectively,and the cloning number of BL-CFC was quantified,as well as Flk-1+ expression was observed by immunofluorescence staining.Positive number and average absorbance were analyzed using IMAGE-PRO PLUS imaging analysis system.RESULTS AND CONCLUSION:Adding bFGF in the course of embryoid body growth could significantly increase the number of BL-CFC(P＜0.01).and the positive results of Flk-1 and the average absorbance were also increased significantly(P＜0.01).bFGF effectively promoted embryoid body amplification and proliferation of BL-CFC.

Objective To investigate the changes of serum osteocalcin and bone micro-structure in ovariectomized mice exposed to low-iron environment.Methods Twenty-four 12-week-old female C57BL/6 mice were divided equally into sham operation (SHAM) group,model(OVX) group,and low iron(OVX+DFO) group.In low-iron group,deferoxamine(DFO) was injected 3 times per week for 5 weeks after operation ; the other groups were injected with the same dose of 0.9％ normal saline for 5 weeks.The serum,left femur,uterus were harvested after five weeks of treatment.The serum osteocalcin and ferritin levels were measured by ELISA kit,the weight of the uterus was recorded by analytical balance.A high resolution micro-CT was used to scan the left femur for cortical bone and cancellous bone analysis.Results (1) The serum osteocalcin and serum ferritin levels in low-iron group were significantly lower than those in the other 2 groups (P＜0.01) ; (2) Compared with the sham group and ovx group,there were significant decrease of the BMD、BV/TV and Tb.N,but increase of Tb.Th and Tb.Sp in low-iron group (P＜0.01).Conclusion A certain dose of DFO (30 mg/kg) can decrease the serum ferritin levels as well as the bone formation index in ovariectomized mice.

AIM: To observe the changes of interleukin-6(IL-6), IL-8 and tumor necrosis factor-?(TNF-?) in serum and lung at different time, and the effects of anisodamine (654-2) treatment in rats with oleic acid-induced ARDS. METHODS: The ARDS model induced by intravenous injection of oleic acid in the rat was used and levels of IL-6, IL-8, TNF-? in serum and lung tissue supernatant were measured using enzyme linked immunosorbent assay (ELISA). RESULTS: Levels of serum and lung tissue IL-6, IL-8, TNF-? in oleic acid type ARDS 4 h group were increased significantly. These cytokines in oleic acid type ARDS 8 h group were lower than that of ARDS 4 h group, but serum IL-6, TNF-? and lung tissue IL-6 were still higher than that of control group . In oleic acid type ARDS 16 h group, serum IL-6, TNF-? were lower than that of the ARDS 8 h group and serum TNF-? and lung tissue IL-6 were higher than that of control group. After 654-2 treatment, the levels of serum and lung tissue IL-6, TNF-? were decreased significantly. CONCLUSION: IL-6, IL-8 and TNF-? might play important roles in the oleic acid-induced ARDS in the rat. 654-2 might alleviate ARDS by inhibiting excess production of IL-6 and TNF-?.

AIM: To observe the dynamic changes of interleukin-4(IL-4),IL-10,IL-12 in rat serum and lung tissues during acute respiratory distress syndrome (ARDS). METHODS: The ARDS model of rats was induced by intravenous injection of oleic acid. The levels of IL-4 ,IL-10,IL-12 in serum and the supernatant of lung tissues were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: The Levels of serum and lung IL-10,IL-12 in ARDS rats were increased in 4 h ,8 h,16 h group compared with control group . The levels in IL-10 in serum in 16 h group and IL-10 in lung tissues of 8 h group were lower than that in 4 h group. The Levels of IL-4 in serum in 4 h, 8 h group were higher than that in control group , while IL-4 in 16 h group was lower than that in 8 h group. IL-4 of lung tissues in 4 h,8 h,16 h group were increased significantly,but in 16 h group were lower than that in 8 h group. The biggest changes of pulmonary coefficient and histopathology were observed at 4 h after injection of oleic acid. CONCLUSIONS: IL-4,IL-10 and IL-12 might play important roles in inflammatory reaction induced by oleic acid. The pro- and anti-inflammatory cytokines produced successively during ARDS. The relationship between unbalanced cytokines and lung injury in ARDS needs to be further studied.

Objective To study the serum levels of progastrin-releasing peptide (pro-GRP) and neuron-specific enolase (NSE) for the clinical diagnosis,therapy monitoring and survival time analysis of small cell lung cancer (SCLC).Methods All 41 SCLC samples (30 males,11 females,age range from 46 to 78 years),95 NSCLC samples (55 males,40 females,age range from 42 to 88 years),and 127 normal individuals samples (80 males,47 females,age range from 35 to 78 years) which were diagnosed by People's Hospital of Zhengzhou from May 1,2008 to April 30,2011 were collected.Serum levels of pro-GRP,NSE and their changes in SCLC patients before and after therapy were evaluated.ANOVA analysis,randomized block design analysis of variance and the log-rank test were collected SPSS 16.0 to evaluate the survival time.Results The serum levels of pro-GRP (median 357.8 ng/L) and NSE (median 89.5 μg/L) in SCLC group were significantly higher than those in the NSCLC group (pro-GRP:39.9 ng/L;NSE:11.43 μg/L) and normal individuals group (pro-GRP:12.7 ng/L;NSE:10.03 μg/L) (P=0.000).The sensitivity of pro-GRP and NSE for the diagnosis of SCLC were 80.4％ and 78.0％,while the specificity were 92％ and 87％,respectively.There is a poor correlation between pro-GRP and NSE serum levels,but when combined the sensitivity can be 95％ and specificity can be 85％.Significantly statistical difference of pro-GRP levels was observed in the different stages of treatment (before and after therapy) in SCLC-LD patients (F =3.53,P =0.038),and significant statistical difference of NSE levels was also observed in SCLC-ED patients in different stages (F =16.049,P =0.000).In partied response SCLC patients,the group with NSE level lower than cut-off value had longer survival time than the other group with NSE level higher than cut-off value (P =0.001).Conclusions The sensitivity of the combined analysis of pro-GRP and NSE is better than single marker for the diagnosis of SCLC.The serum level of pro-GRP has better correlation with therapeutic effect of SCLC-LD patient than NSE.The serum level of NSE are well correlated with therapeutic effect in SCLC-ED patients.There are some certain value of NSE level for evaluation the survival time of SCLC patients who were in partial response.

Objective To explore the clinical value of dual-phase 99Tcm-MIBI scintigraphy in the localization and diagnosis of secondary hyperparathyroidism (SHPT).Methods A total of 20 patients (8 males,12 females; average age 49.6 years) with uremic SHPT who underwent parathyroidectomy from 2010 to 2013 were retrospectively analyzed.All patients underwent 99Tcm-MIBI SPECT/CT and 19 underwent color Doppler ultrasonography (CDUS).Post-excisional histopathology was considered as the gold standard.The diagnostic efficacies of 99Tcm-MIBI and CDUS for SHPT were calculated.The correlation between T/NT ratio in delayed imaging and the volume of excised parathyroid and the intact PTH (iPTH) were analyzed.x2 test,Pearson or Spearson correlation analysis were used to analyze the data.Results The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 99Tcm-MIBI SPECT/CT and CDUS in the diagnosis of SHPT were 66.67％ (44/66),100％(14/14),100％ (44/44),38.89％(14/ 36),72.50％ (58/80) and 78.19％(43/55),52.38％(11/21),81.13％(43/53),47.83(11/23),71.05％ (54/76),respectively.There were significant differences in specificity and positive predictive value (x2 =9.33,9.26,both P＜0.05),but no significant differences in the sensitivity,negative predictive value and accuracy (x2 =1.97,0.04,0.46,all P＞0.05).T/NT ratio correlated with serum iPTH and parathyroid volume (r=0.638,rs =0.571,both P＜0.05).Conclusions The specificity of 99Tcm-MIBI SPECT/CT is superior to CDUS in the diagnosis of SHPT.Dual-phase 99Tcm-MIBI SPECT/CT could locate the hyperfunctional parathyroid gland and provide the basis for surgical treatment.

OBJECTIVETo injvestigate the cerebral blood flow of patients with early syphilis.

METHODS(99)Tc(m)-ECD as brain perfusion imaging agent was used in single photon emission computed tomography (SPECT) for 32 patients with early syphilis and 15 controls. Visual analyses were made on every BSPECT image.

RESULTSThe 32 patients with early syphilis had general, patchy hypoperfusion of cerebral blood flow. Fourteen of the 32 patients had 48 episodes of marked patchy hypoperfusion of rCBF. The responsible areas of hypoperfusion in a patchy distribution involved the left frontal lobe (6 episodes), right frontal lobe (3), left parietal lobe (7), right parietal lobe (6), left temporal lobe (11), right temporal lobe (5), left occipital lobe (3), left basal ganglia (3), cerebellum (1), and nerve nuceus (1). No abnormality was found in the control group.

CONCLUSIONSCerebral blood flow abnormalities exist in patients with early syphilis. General patchy hypoperfusion on SPECT imaging is common.

Adult ; Brain ; diagnostic imaging ; Cerebrovascular Circulation ; physiology ; Humans ; Middle Aged ; Regional Blood Flow ; Syphilis ; diagnostic imaging ; Tomography, Emission-Computed, Single-Photon

Objective To investigate the effects of anti?PD?L1 monoclonal antibody and EGFR?TKI on expression of soluble PD?L1 and function of T lymphocytes in EGFR?mutated lung cancer cells. Methods Flow cytometry was used to analyze the expression of membrane PD?LI. ELISA was performed to detect the level of sPD?L1 in the supernatant of cultured EGFR?mutated and wild type lung cancer cells before and after erlotinib treatment. After treated with anti?PD?L1 monoclonal antibody alone and in combination with erlotinib, the proliferation of T lymphocytes in co?culture system was measured using Cell Counting Kit?8 ( CCK?8) assay. The expression levels of PD?LI and IFN?γin tumor cells and T lymphocytes treated with erlotinib in co?culture system were analyzed by flow cytometry and ELISA, respectively. Results PD?L1 was highly expressed in EGFR?mutated lung cancer PC9 cells (78.7±3.1)% and HCC827 cells (82. 7±2.6)%.After treated with erlotinib, the expression rates of membrane PD?L1 in PC9 and HCC827 cells were down?regulated (64.7%±3.1% and 73.0%±2.6%, respectively),significantly lower than that in the two cell lines without erlotinib treatment ( P<0.05) , and the expression levels of sPD?L1 in the supernatant of PC9 and HCC827 cells were also down?regulated ( 0. 680 ± 0. 120) ng/ml and ( 0. 903 ± 0. 047) ng/ml, respectively, significantly lower than that in the two cell lines without erlotinib treatment ( P<0. 01 ) . However, no significant changes of membrane PD?L1 and sPD?L1 expression were found in EGFR wild type lung cancer cells ( H1299 and A549) before and after erlotinib treatment. In the co?culture system composed of T cells and EGFR?mutated lung cancer cells, treatment with erlotinib alone promoted the proliferation of T lymphocytes (P<0.05), and combined treatment of anti?PD?L1 monoclonal antibody with erlotinib had a stronger effect ( P<0.05) . In the co?culture system composed of T cells and EGFR wild type cell lines, the proliferation of T cells was not changed after using erlotinib alone or combination of erlotinib and anti?PD?L1 monoclonal antibody ( P>0.05) . Before and after treatment with erlotinib, the secretion levels of IFN?γwere (856.0± 70. 3) pg/ml and ( 1 697. 3 ± 161. 0) pg/ml, respectively, showing a significant difference ( P<0.001). The expression rates of membrane PD?L1 were (76.2±0.5)% and (50.9±0.9)%, respectively, also showing a significant difference ( P<0.001) . However, no significant changes in the expression of IFN?γ and membrane PD?L1 were found in the co?culture system composed of T cells and A549 cells. Conclusions Anti?PD?L1 monoclonal antibody combined with EGFR?TKI can effectively promote the proliferation and secretion function of T lymphocytes in the microenvironment of EGFR?mutated lung cancer cells.

Objective To investigate the effects of anti?PD?L1 monoclonal antibody and EGFR?TKI on expression of soluble PD?L1 and function of T lymphocytes in EGFR?mutated lung cancer cells. Methods Flow cytometry was used to analyze the expression of membrane PD?LI. ELISA was performed to detect the level of sPD?L1 in the supernatant of cultured EGFR?mutated and wild type lung cancer cells before and after erlotinib treatment. After treated with anti?PD?L1 monoclonal antibody alone and in combination with erlotinib, the proliferation of T lymphocytes in co?culture system was measured using Cell Counting Kit?8 ( CCK?8) assay. The expression levels of PD?LI and IFN?γin tumor cells and T lymphocytes treated with erlotinib in co?culture system were analyzed by flow cytometry and ELISA, respectively. Results PD?L1 was highly expressed in EGFR?mutated lung cancer PC9 cells (78.7±3.1)% and HCC827 cells (82. 7±2.6)%.After treated with erlotinib, the expression rates of membrane PD?L1 in PC9 and HCC827 cells were down?regulated (64.7%±3.1% and 73.0%±2.6%, respectively),significantly lower than that in the two cell lines without erlotinib treatment ( P<0.05) , and the expression levels of sPD?L1 in the supernatant of PC9 and HCC827 cells were also down?regulated ( 0. 680 ± 0. 120) ng/ml and ( 0. 903 ± 0. 047) ng/ml, respectively, significantly lower than that in the two cell lines without erlotinib treatment ( P<0. 01 ) . However, no significant changes of membrane PD?L1 and sPD?L1 expression were found in EGFR wild type lung cancer cells ( H1299 and A549) before and after erlotinib treatment. In the co?culture system composed of T cells and EGFR?mutated lung cancer cells, treatment with erlotinib alone promoted the proliferation of T lymphocytes (P<0.05), and combined treatment of anti?PD?L1 monoclonal antibody with erlotinib had a stronger effect ( P<0.05) . In the co?culture system composed of T cells and EGFR wild type cell lines, the proliferation of T cells was not changed after using erlotinib alone or combination of erlotinib and anti?PD?L1 monoclonal antibody ( P>0.05) . Before and after treatment with erlotinib, the secretion levels of IFN?γwere (856.0± 70. 3) pg/ml and ( 1 697. 3 ± 161. 0) pg/ml, respectively, showing a significant difference ( P<0.001). The expression rates of membrane PD?L1 were (76.2±0.5)% and (50.9±0.9)%, respectively, also showing a significant difference ( P<0.001) . However, no significant changes in the expression of IFN?γ and membrane PD?L1 were found in the co?culture system composed of T cells and A549 cells. Conclusions Anti?PD?L1 monoclonal antibody combined with EGFR?TKI can effectively promote the proliferation and secretion function of T lymphocytes in the microenvironment of EGFR?mutated lung cancer cells.