0 05). The level of IL-12 in PB serum was higher than that of PB serum( P

Objective To investigate the expression of forkhead box protein P1(FOXP1)in gastric mucosa-associated lymphoid tissue(MALT)lymphomas and its relationship with histological morphology and prognosis. Methods In this study.samples of 43 MALT lymphoma were studied histologically and divided into monomorphic histology group and polymorphic histology group according to their cellular features.The expressions of FOXP1 and NF-κB in gastric MALT lymphoma were evaluated immunohistochemically by two-step method of Envision,and the clinicopathological features and prognosis were analyzed retrospectively.Results The nuclear expressions of FOXP1 in 43 cases with gastric MALT lymphoma were 44%(19 of 43),including strong immunoreactivity in 7 cases and moderate immunoreactivity in 12 cases.There were 4 cases with positive immunoreactivity in moninorphic histology group and 15 cases in polymorphie histology group,and the difference was statistically significant(15%vs.88%,P＜0.01).All the postoperative recurrent cases were strongly positive with FOXP1 stain,and it was closely with FOXP1 expression(P＜0.01).The median survival time(26 months)in polymorphic histology group was significantly shorter than that(123 months)in monmorphic histology group(P＜0.01),and the median survival time was significantly longer in negative FOXP1 expression group than that in moderate FOXP1 expression group and in strong FOXP1 expression group(115 vs.55 vs.12 months)(P＜0.05).similarly,the median survival time in nuclear factor kappa B(NF-κB)expression group was significantly shorter than that in negative NF-κB expression group(26 vs.131 months)(P＜0.01).The median survival time in stageⅠ(98 months)and stage Ⅱ(121 months)was significantly longer than that in stage Ⅱ E+Ⅳ(33 months)(P＜0.01).By multivariate COX regression analysis.FOXP1 nuclear expression and clinical stage were independently prognostic factom. Conclusion FOXP1 expression may be used as a biomarker for the assessment of malignant transformation to diffuse large B-cell lymphoma(DLBCL)and predicting prognosis.

Objective To explore the Clinicopathological characteristics of patients with primary adenosquamous and squamous carcinoma of stomach. Methods The clinical data of 12 cases of primary squamous and adenosquamous carcinoma of the stomach were reviewed retrospectively, and the immunohistochemical staining of CK17 and CKI8 protein were performed in primary gastric adenosquamous carcinoma. Results Primary adenosquamous and squamous carcinoma of the stomach accounted for 0.28% of all the 4352 patients with gastric cancer during the same period. Of the 12 patients, 10 were adenosquamous carcinoma and other two were squamous carcinoma. There were 10 males and 2 females in this group, with their mean age being 65 years. The main clinical presentation included epigastric pain and discomfort, followed by hematemesis and melena. The definite diagnosis rate was 33% (4/12) by gastroscopy and biopsy before operation. The tumors were less than 5 cm in diameter in 3 patients, and more than 5 cm in 9 patients. The surgical procedure was radical resection in 8 patients and palliative resection in 4 patients. There were 1 case of stage Ⅰ, 5 cases of stage Ⅲ, 6 cases of stage Ⅳ. 10 patients died of tumor recurrence and metastasis within 2 years after operation, one died of other unrelated disease, and one was alive for more than 5 months. The component of both adenosquamous and squamous carcinoma were more than 30% in 4 patients with adenosquamous carcinoma who underwent palliative resection and died within 6 months after operation. Conclusions Primary adenosquamous and squamous carcinoma of the stomach were rare, and had specific clinicopathological characteristics. Having both biological behaviours of adenocarcinoma and squamous carcinoma may lead to poor prognosis in adenosquamous carcinoma of stomach.

Objective To investigate the expressions of CD133 and CD44 and their prognostic significance in gastrointestinal stromal tumors (GIST).Methods Streptavidin perosidase (SP) method of immunohistochemistry was used to detect expressions of CD133 and CD44 proteins in 42 cases of GIST, and the relationship between their expressions and tumor size, mitotic count were analyzed by univariate and multivariate factor analyses.Results The expressions of CD133 and CD44 proteins in GIST were 21.4％ (9/42) and 78.6％ (33/42), respectively.The expressions of CD133 and CD44 proteins were significantly correlated with tumor size and mitotic count (P ＜ 0.05).Univariate factor analysis showed that the overall survival of GIST patients with positive CD133 protein (23.2 months) was shorter than that of patients with negative CD133 protein(63.1 months) (P ＜ 0.05).The overall survival of GIST patients with negative CD44 protein (23.2 months) was shorter than that of patients with positive CD44 protein (63.3 months) (P ＜ 0.05).Multivariate factor analysis showed that tumor size, mitotic count and CD44 protein were independent prognostic indicators for survival time after operation.Conclusions The positive expressions of CD133 and CD44 proteins might be the prognostic factors of GIST patients.

Objective: To study the extraction technology of ursolic acid from sambucus chinensis Lindl. Methods: The method of ethanol extraction and agglutination separation was adopted for extracting ursolic acid. Results: The extraction rate was 90%, its purification was 98%, the product was recognized to be ursolic acid by physicochemical contents and spectral identification. Conclusion: This method is advanced, practical, reasonable and feasible. It can be applied in industrial production.

AIM: In order to replace mouse embryonic fibroblast cells by human embryonic fibroblast cells to support the growth of human embryonic stem cells, the effects of human/mouse embryonic fibroblast cells on the growth of human embryonic stem cells were compared. METHODS: Both mouse and human embryonic fibroblast cells were used as feeder layer to support human embryonic stem cells. The proliferation and differentiation of human embryonic cells were observed. RESULTS: Combined use of human leukemia inhibitory factor with human/mouse feeder layer cells would support growth and proliferation of human embryonic stem cells and kept in undifferentiated condition. There was no difference between human/mouse cell. CONCLUSION: Human embryonic fibroblast cells can be used to support proliferation of human embryonic stem cells, eliminating the influence of foreign protein.

BACKGROUND:It has been reported that treatment with granulocyte-colony stimulating factor (G-CSF) increases the abundance of circulating CD34+ cells in rats. Data from the study, more important, suggested that mobilized by G-CSF may enhance rapid reendothelization and reduce neointimal formation after vascular injury.OBJECTIVE: To evaluate whether BM-derived CD34+ cells could enhance rapid reendothelization and reduce neointimal formation after balloon-injured carotid artery in an intact rat model.DESIGN: Randomized control animal study.SETTING: Nanjing First Hospital Affiliated to Nanjing Medical University.MATERIALS: The experiment was carried out in Nanjing First Hospital from December 2005 to April 2006. A total of 40 male Sprague-Dawley rats, weighing 200-250 g and of SPF grade, were purchased from National Rodent Laboratory Animal Resources, Shanghai Branch. The recombinant human G-CSF was purchased from Qilu Pharmaceutical. The 2F Fogarty arterial embolectomy catheters were purchased from Edwards Lifesciences. Anti-human CD34 and anti-human CD45 were purchased from Multi Sciences.METHODS: SD rats were divided randomly into treated group (n =20) and control group (n =20). Subcutaneous injection of recombinant human G-CSF (100 μg/kg/day) once daily for 8 days for treated group. Control group as treated with subcutaneous injection of saline. Five days after initiation of G-CSF treatment or saline, the rats were anesthetized by intraperitoneal injection with ketamine. The left common carotid artery was exposed through a midline incision of the ventral side of the neck. A 2F Fogarty arterial embolectomy catheter was inserted through the external carotid artery,inflated with 200 μL air, and passed 3 times along the length of the segment, which was defined proximally by the carotid bifurcation and distally by the edge of the omohyoid muscle. After removal of the catheter, the proximal ligature of the external carotid artery was tied off. ① An average of 1 mL venous blood per rat was collected for enumeration of the white blood wells (WBCs) and CD34+ cells before and 5 days after initiating G-CSF or saline treatment. ② Ten rats in each group were killed with overdose ketamine at 14 and 28 days after balloon injury and left common carotid arteries were harvested. The luminal surface of carotid arteries (n =5, each group) was exposed to calculate the reendothelialized area, which was manually traced with software (Image ProPlus). Reendothelialized area = non-stained with Evans blue area/the total area of balloon-injuried. The cross sections of carotid arteries (n =5, each group) were stained with hematoxylin and eosin (HE) and calculated intima-to-media area ratio (I/M) with software (Image ProPlus) to assess the extent of neointimal thickening. ③ To evaluate the extent of reendothelialization of arteries injury, sections were stained with CD31 and vWF by immunohistochemistry analysis.MAIN OUTCOME MEASURES: ① The number of WBCs and CD34+ cells; ② the extent of reendothelialization of arteries injury; ③ the extent of neointimal hyperplasia (I/M); ④ CD31 + and vWF+ endothelial cells.RESULTS: A total of 40 rats were involved in the final analysis. ① The number of WBCs and CD34+ cells: After 5 days of treatment, the number of WBCs in the treated rats increased more than 2.7-fold compared with control group [(27.60±2.45) ×109 L-1, (10.11±1.81) ×109 L-1, P ＜ 0.01], CD34+ cells increased more than 12.2-fold compared with control group (38.31×107 L-1, 3.14×107 L-1, P ＜ 0.01). ② The extent of reendothelialization: At 14 and 28 days after balloon injury,carotid artery of reendothelialization in the treated group were (68.3±8.3)％ and (97.6±4.1)％, superior than the control group (33.8±6.3)％ and (76.1±5.2)％ (P ＜ 0.01). ③ The extent of neointimal hyperplasia: At 14 and 28 days after balloon injury, the neointima-media (I/M) ratios in the treated rats were 0.39±0.11 and 0.45±0.09, less than the control group 0.87±0.15,1.26±0.16 (P ＜ 0.01). A highly significant inhibition of neointimal hyperplasia was observed in the treated group. ④ CD31+ and vWF+ endothelial cells: At 28 days after injury, sections from G-CSF treated group showed almost complete and continuous monolayer of CD31 and vWF positive cells.In contrast, a patchy and interrupted CD31 and vWF positive cells were found lining the lumen of control group.CONCLUSION:Treatment with G-CSF significantly increases the number of CD34+ cells and accelerates the rate of reendothelialization of injured vessels, leading to marked inhibition of neointimal formation after vascular injury in rats.

[Objective]To investigate the effects of mesenchymal stem cells (MSC) feeder layer, culture sere and freeze-thaw lysates on expansion and differentiation of cord blood CD34~+ cells in vitro. [Methods] MSC were isolated from human bone marrow and cultured until the third passage. Sera were obtained from the cultured MSC, and freeze-thaw lysates were obtained by repeated freeze-thaw procedures. Cord blood CD34~+ cells were isolated by magnetic cell separation system, and were co-cultured with the MSC feeder layer, culture sera, freeze-thaw lysates and hematopeietic growth factors (HGFs), respectively. The nucleated cells, CD34~+ cells, CD34~+CD38~- cells, CD41~+ cells and CD3~+ cells in the above culture system were detected by flow cytometry on day 6 and day 12. [Results] ①MSC feeder layer had a strong effect on nucleated cells, CD34~+,CD34~+CD38~- cells expansion. The MSC sera and freeze-thaw lysates had similar effect on cell expansion, but the effect was weaker than that of feeder layer (P<0.05). ② Both MSC sera and feeder layer inhibited cord blood CD34~+ cells differentiation toward CD3~+ cells or CD19~+ cells, and no significant differences were found between these two groups (P>0.05). ③ Both MSC sera and feeder layer promoted cord blood CD34~+ cells differentiation toward CD41~+ cells, and the effect was stronger in the feeder layer than that of the sera (P<0.05). ④ Freeze-thaw lysates had no effect on cell expansion and differentiation, and were similar with that of HGFs (P>0.05). [Conclusions] The MSC sera have positive effects on expansion of cord blood CD34~+ and CD34~+CD38~- cells, moreover they have the ability of promoting cord blood CD34~+ cells differentiation toward CD41~+ cells.

AIM and METHODS: To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group. RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13. CONCLUSION: Measles vaccine could down-regulate IL-13 level, therefore decrease total IgE synthesis, but not affect IL-12 level in asthmatic children.

AIM: To investigate the efficacy of dendritic cells (DCs) that augments the cytotoxic activity of cytokine-induced killer (CIK) cells, natural killer (NK) cells from a same donor and the CD45RO expression on CIK cells. METHODS: The expanded killer cells were divided into two groups: group A was pre-cocultured with DCs for 6 days, group B was the control that without any stimulation. Cytotoxicity of CIK and NK cells was measured at different effect-target ratio against K562 and HL-60. CD45RO and CD45RA expression on CIK cells in different groups were detected by flow cytometry. RESULTS: Cytotoxicity of CB derived killer cells was positive correlation with effect-target ratio. The cytotoxicity of group A against HL-60 was higher than that of group B significantly. At 20∶1 effector-target ratio, the lytic activity of group A CIK, NK cells against K562 was higher than that of group B significantly, but no significant difference between them at 10∶1 effector-target ratio. The CD45RO expression on CIK cells in groups A was significantly higher than that in groups B. CONCLUSION: CIK and NK cells cocultured with DCs can augment the killer's cytotoxicity against tumor cells and promote the CD45RO expression on CIK cells.