AIM: To observe the expression of β-catenin in patients with chronic myeloid leukemia (CML) at different disease phases, and to analyze the relationship between BCR-ABL and cytogenetic response to imatinib mesylate. METHODS: RT-PCR and Western blotting were used to detect β-catenin mRNA and protein expression in bone marrow mononuclear cells (BMMNCs) from 99 patients with CML. The association with BCR-ABL and BCR-ABL fusion was determined by FISH in 94 patients after one year treatment with imatinib mesylate, and the relationship between β-catenin and cytogenetic response to imatinib mesylate was analyzed. RESULTS: The expression of β-catenin was increased significantly in patients with blast crisis and accelerated phase (P<0.01), while the expression of β-catenin between normal person and chronic phase of CML patients was not statistically different (P>0.05). No significant relation between β-catenin and BCR-ABL expression (r=0.314, P>0.05) was observed. The expression of β-catenin was increased significantly in the patients who did not reach main cytogenetic remission (P<0.01). CONCLUSION: The patients in progression phases of CML over-express β-catenin. The expression of β-catenin is not significantly related to BCR-ABL expression, but related to the therapeutic response of imatinib. Beta-catenin may be involved in the mechanism of CML progression and could be used as a new therapeutic target.

AIM: To investigate the effects of in situ transplantation of mobilized autologous bone marrow stem cells on infarction size and cardiac function in patients with acute myocardial infarction. METHODS:25 patients with first acute myocardial infarction were randomly divided into stem cells in situ transplantation group and control group, 12 patients in stem cells in situ transplantation group were injected subcutaneously with 300 ?g granulocyte colony-stimulating factor(G-CSF) daily for four days in addition to standard therapy. 13 patients in control group were treated with standard therapy alone. The conventional 12 leads electrocardiogram were recorded on 1, 28 days after admission and the cardiac function was scored by the QRS scoring system proposed by Wagner. Furthermore, the infarction size was assessed by radionuclide myocardial perfusion imaging 7, 28 days after admission. RESULTS:4 weeks after admission, the QRS scores decreased, the infarction size reduced significantly in the stem cells in situ transplantation group (from 36.0%?8.3% to 18.0%?5.8%, P

Objective To observe the clinical efficacy of Wendan decoction in the treatment of phlegm-heat accumulation type acie vulgaris.Methods 62 patients with phlegm-heat accumulation type ache vulgaris were randomly divided into a treatment group and a control group randomly,with 31 patients in each group.The control group was given dietary advice and skin cleansing techniques,while the treatment group was given oral administration of Wendan decoction on the basis of the control group.Results ①skin damage comparison:after the treatment,skin damage symptoms such as tenderness (0.71 ±0.32),redness (0.47±0.63),cyst(0.59±0.53),and tubercle (0.63±0.54) in the treatment group were significantly improved than the control group [the values were (1.12±0.32),(1.44±0.21),(1.40±0.36),(1.18±0.43),respectively] (P＜0.05).②symptoms score comparison:The symptoms score in the treatment group after the treatment (42.95±1.53) was significantly improved than that before the treatment (52.89± 1.98),(P＜0.05),and also obviously better than the control group after the treatment.③clinical efficacy comparison:Total efficacy was 96.77％ in the treatment group and 54.83％ in the control group,showing marked difference(P＜0.05).Conclusion Wendan decoction is effective in treating phlegm-heat accumulation type acne vulgaris.

AIM:To investigate effects of thrombopoietin(TPO) and TPOⅡ on human platelet activation in vitro. METHODS:Human platelets were incubated in the phosphate-buffered saline containing rhTPO or TPOⅡ at the concentration of 100 μg/L for five minutes. In order to determine the rate of platelet activation. The CD62P and CD41 expressions on platelets were analysed by flow cytometry using fluorescence labelled monoclonal antibody to CD62P and CD41. RESULTS:The results demonstrated that expression of CD62P on platelets which were incubated with rhTPO or TPOⅡ didn't increase compared with that of contrast group. CONCLUSION:Both rhTPO and TPOⅡdidn't cause the disorder of platelet activation.

OBJECTIVE To study the relationship of the expression of PLOD2 protein in laryngeal carcinoma and the clinicopathological features of patients. METHODS The expression of PLOD2 in paraffin-embedded specimens of 114 patients with laryngeal carcinoma was detected by immunohistochemistry. The relationship between the expression of PLOD2 and clinicopathological features was analyzed by χ2 t est, s urvival a nalysis b y K aplan-Meier method, and multivariate analysis of Cox proportional hazard model. The fresh frozen specimens of 8 patients randomly selected from the patients were detected by realtime quantitative polymerase chain reaction and Western blotting for the expression of PLOD2 in tumor tissues and adjacent normal tissues. RESULTS PLOD2 protein was associated w ith c linical s tage a nd T s tage(P <0.05). The expression level of PLOD2 protein in laryngeal squamous cell carcinoma was higher than that in adjacent normal tissue(P <0.05). Kaplan-Meier survival analysis showed that low expression of PLOD2 was associated with patient survival rate(χ2=12.484, P <0.001). Multivariate Cox regression analysis showed that PLOD2 protein expression and M stage were independent risk factors for laryngeal cancer growth (P value, both <0.05). CONCLUSION The level of POLD2 protein expression was positively correlated with clinical stage and T stage. PLOD2 protein is an independent risk factor for the growth of laryngeal cancer. The higher the expression of PLOD2 protein, the lower the prognosis of patients. PLOD2 protein expression may play an important role in the growth and prognosis of laryngeal cancer, and may be a new molecular marker for judging the growth and prognosis of laryngeal cancer.

AIM: To investigate the biological activity of thrombopoietin Ⅱ(TPOⅡ) in vivo , which consists of two new kinds of ligand binding with thrombopoietin receptor. METHODS: Purified ligandⅠof TPOⅡ, artificial compound ligandⅡ of TPOⅡand rhTPO were injected into purebred Babl/c mice respectively in 7 days by intraperitoneal injection once for a day. Then the biological activity of TPOⅡ was analyzed by measuring peripheral platelet counts by the end of the seventh day. RESULTS: On the seventh day, the platelet counts of mice treated by ligandⅠof TPOⅡ were higher than that in the negative control group( P 0.05). On the fourteenth day, the platelet counts increased in two all experimental groups of TPOⅡcompared with negative control group( P 0.05). Moreover the platelet counts of mice in two experimental groups of TPOⅡ and the positive group showed increase with experimental days. CONCLUSION: The purified ligandⅠof TPOⅡ had obvious activity in increasing platelet production, which is not different from the effect of rhTPO.

AIM: To investigate effects of thrombopoietin(TPO) and TPOⅡ on human platelet activation in vitro. METHODS:Human platelets were incubated in the phosphate-buffered saline containing rhTPO or TPOⅡ at the concentration of 100 ?g/L for five minutes. In order to determine the rate of platelet activation. The CD62P and CD41 expressions on platelets were analysed by flow cytometry using fluorescence labelled monoclonal antibody to CD62P and CD41. RESULTS: The results demonstrated that expression of CD62P on platelets which were incubated with rhTPO or TPOⅡ didn't increase compared with that of contrast group. CONCLUSION: Both rhTPO and TPOⅡdidn't cause the disorder of platelet activation.

Objective To explore the correlation of urinary cysteinyl leukotriene F4 (CysLTE4) and diagnosis of bronchopulmonary dysplasia (BPD) in premature infants.Method One hundred and fifty-eight newborn infants were consecutively admitted to the neonatal intensive care units of First Affiliated Hospital of Nanjing Medical University from November 2014 to October 2015.The infants were divided into 3 groups according to the diagnosis on discharge.Sixty-one term infants were classified as having no pulmonary diseases,52 premature infants were classified as without BPD,and 45 premature infants with BPD were diagnosed at 28 d after birth.Urinary CysLTE4 levels of newborns within 3 days after birth were measured in a blinded way by enzyme-linked immunosorbent assay and were compared among 3 groups,and were evaluated for the diagnostic value and the correlation of gestational age and birth weight.Statistical analysis was performed using correlation analysis,one-way analysis of variance and x2 test etc.Result In infants with BPD,the mean urinary CysLTE4 level was (191.0 ± 29.3) ng/L which significantly higher than the premature group without BPD ((164.1 ±22.7) ng/L) and term infant group ((151.6 ±41.9) ng/L,F =18.70,P ＜ 0.05).Urinary CysLTE4 level within 3 days of life in newborn inversely correlated with gestational age and birth weight (Pearson =-0.33,-0.38,P ＜ 0.01).The area under the curve was 0.78,95％ CI:0.70-0.86,P ＜0.01,when cutoff was 187.7 ng/L,with Youden index 0.59,sensitivity 77.8％ and specificity 81.4％,respectively.Conclusion Urinary CysLTE4 level is up-regulated in BPD infants within early days of life which may be a useful biomarker of early diagnoses of BPD.

Objective:To investigate the effects and mechanisms of allogeneic epidermal stem cells (ESCs) on the survival of allogeneic full-thickness skin grafts in nude mice with full-thickness skin defect wounds.Methods:Experimental research methods were applied. Primary ESCs that appeared paving stone-like after being cultured for 7 d were obtained by enzymatic digestion method from one 4-week-old male BALB/c-NU nude mouse (the same strain, age, and sex below). The cells of third passage were identified by flow cytometry to positively express ESC marker CD44 and negatively express CD45, meanwhile, the positive expression of ESC markers of p63 and integrin 6α, and negative expression of CD71 were identified by immunofluorescence method. The ESCs of third passage in the logarithmic growth phase were used for the following experiments. Twenty-six nude mice were equally divided into phosphate buffered saline (PBS) group and ESCs group according to the random number table. A full-thickness skin defect wound was made on the back of each nude mouse, and then the wounds of the two groups were sprayed with equal volumes of PBS and ESCs, respectively. The wounds were transplanted with full-thickness skin grafts cut from the backs of 4 other nude mice. Each ten nude mice from the two groups were selected, the wound healing and skin survival on post surgery day (PSD) 0 (immediately), 3, 7, 14, and 21 were observed, and the survival ratio and shrinkage rate of skin grafts on PSD 3, 7, 14, and 21 were calculated (the number of sample was the number of surviving skin grafts at each time point); the blood perfusion in the skin grafts on PSD 3, 7, and 14 was detected by the laser speckle blood flow imager, and the blood flow ratio of nude mice skin grafts in ESCs group to PBS group at each time point was calculated (the number of sample was the pair number of surviving skin grafts in group pairing at each time point). The skin graft tissue of each 3 nude mice remained in the two groups were collected on PSD 7, and the mRNA expressions and protein expressions of tumor necrosis factor α (TNF-α), interleukin 8 (IL-8), IL-10, type Ⅰ collagen, type Ⅲ collagen, and matrix metalloproteinase 9 (MMP-9) in the tissue were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction and Western blotting, respectively. Data were statistically analyzed with Log-rank test, analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and Bonferroni correction. Results:Taking the condition on PSD 0 as a reference, the wounds of nude mice in the two groups healed gradually on PSD 3, 7, 14, and 21, and the shrinkage of skin grafts was gradually obvious. Among them, the shrinkage healing of wound of nude mice in PBS group was more significant than that in ESCs group. On PSD 3, the skin graft of 1 nude mouse failed in ESCs group, while the skin graft of 3 nude mice failed in PBS group. On PSD 7, the skin graft of another nude mouse failed in PBS group. The survival ratio of skin grafts of nude mice in the two groups was similar on PSD 3, 7, 14, and 21 ( P>0.05). On PSD 3, 7, 14, and 21, the shrinkage rates of skin grafts of nude mice in ESCs group were (9.2±0.4)%, (19.7±1.2)%, (53.6±3.5)%, and (62.2±5.1)%, respectively, which was significantly lower than (11.0±0.9)%, (47.8±2.8)%, (86.1±7.1)%, and (89.7±9.0)% in PBS group ( t=5.719, 26.650, 11.940, 7.617, P<0.01). On PSD 3, 7, and 14, blood perfusion signals were observed in the skin grafts of nude mice in the two groups. The average blood perfusion ratios of the skin grafts of nude mice in ESCs group to PBS group were greater than 1, and there was no statistically significant difference in the overall comparison of 3 time points ( P>0.05). On PSD 7, compared with those of PBS group, the mRNA and protein expressions of TNF-α, IL-8, type Ⅰ collagen, and type Ⅲ collagen in the skin graft tissue of nude mice in ESCs group were significantly reduced, while the mRNA and protein expressions of IL-10 and MMP-9 in the skin graft tissue of nude mice in ESCs group were significantly increased (in mRNA comparison, t=2.823, 2.934, 2.845, 2.860, 3.877, 2.916, P<0.05). Conclusions:Allogeneic ESCs can reduce the shrinkage of allogeneic full-thickness skin grafts transplanted on full-thickness skin defect wounds in nude mice, promote the formation of new blood vessels between the skin graft and the wound, reduce inflammation and collagen protein expression, and promote the expression of MMP-9, thus improving the survival quality of skin grafts.