This study was purposed to investigate the effect of aminopeptidase N/CD13 on bestatin enhancing all-trans-retinoic acid (ATRA)-inducing differentiation in NB4 cells. The nitroblue-tetrazolium (NBT) reduction assay was performed to determine the differentiation of NB4 cells, MR2 cells and primary APL blasts. The expression of P38 MAPK protein and the phosphorylation of P38 MAPK protein in NB4, MR2 and K562 cells were detected by Western blot. The results showed that pre-incubation with 5 µg/ml WM-15 blocked the enhancement effect of bestatin on differentiation of NB4 cells induced by ATRA. 5 µg/ml CD13 antibody WM-15 partly blocked the inhibition of bestatin on the phosphorylation of P38 MAPK in NB4 cells. 100 µg/ml bestatin inhibited the phosphorylation of P38 MAPK in NB4 cells and MR2 cells in a time-dependent manner. 100 µg/ml bestatin had no effect on the phosphorylation of P38 MAPK in K562 cells with low level of CD13. Bestatin could not restore the sensitivity to ATRA in ATRA-resistant primary APL blasts and MR2 cells. It is concluded that aminopeptidase N/CD13 inhibitor bestatin may enhance the differentiation-inducing activity of ATRA through inhibiting the phosphorylation of P38 MAPK in NB4 cells mediated by the cell surface APN/CD13.
Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; CD13 Antigens ; antagonists & inhibitors ; metabolism ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Humans ; Leucine ; analogs & derivatives ; pharmacology ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Phosphorylation ; Tretinoin ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effects of interleukin 7 (IL-7) on B7 molecules expression and immunogenicity of acute leukemia (AL) cells.

METHODSThe B7 molecules expression on fresh acute leukemia cells and on the IL-7 exposed leukemia cells was detected by FACScan cytometer. B7-1 and B7-2 mRNA in IL-7 treated HL-60 cells were detected by reverse transcription-PCR (RT-PCR). The stimulation of proliferation of allogeneic peripheral blood mononuclear cells (PBMNC) by IL-7 treated leukemia cells was detected by MTT method. The level of interferon-gamma (IFN-gamma) secreted by the stimulated PBMNC was determined using enzyme-linked immunosorbent assays (ELISA). The blocking experiments were performed using monoclonal antibodies against B7-1, B7-2 and W6/32.

RESULTSB7-1 was weakly expressed in three, whereas B7-2 did in only one of eleven AL patients. IL-7 significantly enhanced B7 molecules expression on AL cells in a time-dependent manner. Furthermore, IL-7 could induce higher expression of B7-1 and B7-2 mRNAs on HL-60 cells. IL-7 treated leukemia cells could stimulate PBMNC proliferation and promote their IFN-gamma production. Anti-B7-1 and anti W6/32 but not anti-B7-2 monoclonal antibodies significantly inhibited the stimulated PBMNC proliferation and IFN-gamma secretion.

CONCLUSIONFresh AL cells express low level of B7-1 and B7-2 molecules. IL-7 enhances the B7 molecules expression on AL cells. The IL-7-treated leukemia cells can significantly stimulate the proliferation of allogeneic PBMNC and induce their IFN gamma secretion.

B7-1 Antigen ; genetics ; metabolism ; B7-2 Antigen ; genetics ; metabolism ; Humans ; Interleukin-7 ; pharmacology ; Leukemia ; immunology ; metabolism ; RNA, Messenger ; genetics ; Tumor Cells, Cultured ; Up-Regulation ; drug effects

Objective To investigate upper esophageal sphincter (UES)abnormalities in patients with achalasia (AC),and to analyze the correlation between UES abnormalities and clinical symptoms, treatment efficacy.Methods From February 2012 to December 2014,158 patients with AC and received high resolution manometry (HRM)examination were retrospectivly analyzed.According to whether with UES abnormalities,patients were divided into UES normal group and UES abnormal group.Patients of UES abnormal group were sub-divided into UES hypotensive group (UES resting pressure<34 mmHg, 1 mmHg=0.133 kPa),hypertensive group (UES resting pressure>104 mmHg)and impaired relaxation group (residual pressure>12 mmHg).Analysis of Variance,Kruskal-Wallis H test and Chi square test were performed to compare the clinical data and dynamic characteristics of the patients in each group. Results A total of 74 (46.8%)AC patients had UES abnormalities,the majority of whom were impaired relaxation (35 cases,47.3%).The age of patients in hypotensive group ((60.6 ± 10.1 )years)was significantly older than that of hypertensive group ((43.9 ±11 .1 )years)and impaired relaxation group ((46.8±16.3)years),and the disease course (10 years,4 to 30 years)was obviously longer than that of hypertensive group (6 years,1 to 10 years)and impaired relaxation group (8 years,3 to 15 years),and the differences were statistically significant (F = 7.983,H = 13.816,both P < 0.01).There was no correlation between UES abnormalities and clinical symptoms (P >0.05 ).The results of AC subtyping indicated that type Ⅱ AC accounted 55 .7% (88/158).Type Ⅱ AC cases number of UES normal group and abnormal group was 46 and 42 cases,both was majority (54.8% and 56.8%).Among these patients,123 patients finally received peroral endoscopic myotomy (POEM),47.2%(58/123 )of whom had abnormal UES.More than 85 % patients were satisfied at one month after the operation.And Eckardt scores significantly decreased.There was no significant difference in treatment efficacy between the two groups.Conclusions Most AC patients are with UES abnormality,and impaired relaxation is more common.There is no correlation between UES abnormalities and major symptoms.There is no predictive role of UES abnormalities in treatment efficacy of POEM in AC patients.

Objective To investigate and evaluate the biomechanical property of the skin in pig's back in order to provide the essential theoretical basis for clinical and skin products.Method Taking the skin in pig's back as experimental material,the monotonic tensile and cyclic tension-tension tests with difierent loading rates was researohed respectivaly.Meanwhile,with different loading directions and stress levels the creep and cy-clic tension-tension tests were also been studied experimentally.Result The capacity of resisting tensile,creep and cyclic deformation of pig's skin in the direction along the Langer's line is stronger than that perpen-dicuiar to the Langer's line.The creep curve of pig's skin is load-dependent and consisted of three phases a-bout deceleration phase,stabilization phase and destruction stage.Pig's skin exhibits apparent ratcheting un-der asymmetry stress cycle.Ratcheting deformation displays significant mean stress,stress amplitude and loading speed dependence.Condusion Based on the experiment,the biomechanics property of skin's vis-coelasticity and anisotropic feature have been sysmarie stadied,it's provide necessaw theoretical fundation for clinical and leather products.

Objective To explore the value of post processing technique of dual source CT in diagnosing peripheral PE.Methods One hundred and fifty patients with suspected PE were underwent dual source CT scan,and 20 of the all patients met the criterion. The raw CT data were reconstructed by the pulmonary embolism detection (PED)software,double energy perfusion imaging (DEPI)and CT pulmonary artery angiography (CTPA).The PED,DEPI and CTPA map were analyzed by 2 senior doctors in cardiovascular diagnosis profession.According to the diagnosis standard of PE,the location,number of the emboli in segmental and sub-segmental pulmonary arteries were respectively recorded on the PED and DEPI map.We calculated the detection rate,evaluated the significant difference and evaluated the diagnosis consistency between the DEPI and CTPA map.Results Emboli were found in 30 segmental and 40 sub-segmental arteries on the CTPA map with the detection rate of 7.50% and 5.00%,respectively.Emboli were found in 48 segmental and 62 sub-segmental arteries on PED map with the detection rate of 12.00% and 7.75%,respectively.There was significant difference between the CTPA and PED map (χ2 =4.60,5.06,P<0.05).The DEPI and PED map had higher consistency in diagnosing PE.The Kappa coefficient was 0.94 if the PED map was regarded as the reference standard,Among 48 cases with segmental PE detected by the PED map,there were 13 cases of complete filling defects and 35 cases of partial filling defects.Among 13 cases of complete filling defects,there were perfusion defect in 10, and perfusion thin in 3 on the DEPI map.Among 35 cases of partial filling defects,there were perfusion defect in 2,perfusion thin in 29,and perfusion normal in 4 on the DEPI map.Conclusion The PED map combined the DEPI map of dual source CT can significantly improve the diagnosis rate of peripheral PE and has a high clinical value.

Objective To investigate and evaluate the biomechanical property of the skin in pig's back in order to provide the essential theoretical basis for clinical and skin products.Method Taking the skin in pig's back as experimental material,the monotonic tensile and cyclic tension-tension tests with difierent loading rates was researohed respectivaly.Meanwhile,with different loading directions and stress levels the creep and cy-clic tension-tension tests were also been studied experimentally.Result The capacity of resisting tensile,creep and cyclic deformation of pig's skin in the direction along the Langer's line is stronger than that perpen-dicuiar to the Langer's line.The creep curve of pig's skin is load-dependent and consisted of three phases a-bout deceleration phase,stabilization phase and destruction stage.Pig's skin exhibits apparent ratcheting un-der asymmetry stress cycle.Ratcheting deformation displays significant mean stress,stress amplitude and loading speed dependence.Condusion Based on the experiment,the biomechanics property of skin's vis-coelasticity and anisotropic feature have been sysmarie stadied,it's provide necessaw theoretical fundation for clinical and leather products.

The bioactivities, chemical composition and distribution of aerial parts of Panax species are different from the roots. The present paper summarized the phytochemical and analytical studies of aerial parts of Panax species, including P. ginseng, P. notoginseng, P. quinquefoliun and P. japonicus. This review aims so as to provide scientific evidences for further investigation of chemical profile, quality control and optimal utilization of these resources.
Chemistry Techniques, Analytical ; Drugs, Chinese Herbal ; chemistry ; Panax ; chemistry ; Plant Components, Aerial ; chemistry ; Quality Control ; Saponins ; analysis ; chemistry

This study was purposed to investigate whether aminopeptidase inhibitor, bestatin, can potentiate all-trans retinoic acid (ATRA)-inducing differentiation in NB4 cells, and to explore its mechanism. The NB4 cells were exposed to either bestatin and ATRA alone or in combination, the morphological changes of NB4 cells were observed by optical microscopy, the CD11b expression was measured by flow cytometry, the function of defferentiation cells was analyzed by nitroblue-tetrazolium (NBT) reduction assay, the mRNA expressions of c-myc and c-EBPepsilon in NB4 cells were detected by RT-PCR, the c-Myc protein expression was determined by Western blot. The results showed that treatment with bestatin alone induced no significant changes in morphology, NBT reduction activity and CD11b expression in NB4 cells. NB4 cells incubated with 10 nmol/L ATRA plus 100 microg/ml bestatin showed more morphologic feature of metamyelocyte and band neutrophil than ATRA alone treated cells. 100 microg/ml bestatin enhanced the NBT reduction activity in NB4 cells induced by various concentrations of ATRA (10, 20, 40 nmol/L). The effects of various concentrations of ATRA in combination with 100 microg/ml bestatin were statistically different from the effect of ATRA alone (P < 0.01). From 48 to 96 hours, 100 microg/ml bestatin time-dependently increased NBT reduction in NB4 cells induced by 10 nmol/L ATRA (P < 0.01). 10 nmol/L ATRA plus 100 microg/ml bestatin for 72 hours prominently elevated CD11b expression in NB4 cells as compared with ATRA alone treated NB4 cells (P < 0.01). There was a substantial decrease in c-myc mRNA levels when 100 microg/ml bestatin was added to 10 nmol/L ATRA (P < 0.05). Various concentrations (50, 75, 100 microg/ml) of bestatin combined with 10 nmol/L ATRA down-regulated the expression of c-Myc protein, which was negatively correlated with the NBT reduction activity of NB4 cells induced by 10 nmol/L ATRA alone or plus bestatin at various concentrations (r = -0.940, P = 0.017). However, 100 microg/ml bestatin plus 10 nmol/L ATRA could not induce any significant changes in the levels of c-EBPepsilon mRNA as compared with ATRA alone treated NB4 cells. It is concluded that an aminopeptidase inhibitor bestatin can potentiate ATRA-inducing differentiation of NB4 cells, possibly by down-regulating c-myc expression in synergy with ATRA.
Aminopeptidases ; antagonists & inhibitors ; Antibiotics, Antineoplastic ; pharmacology ; Cell Transformation, Neoplastic ; drug effects ; Down-Regulation ; Drug Synergism ; Humans ; Leucine ; analogs & derivatives ; pharmacology ; Leukemia, Promyelocytic, Acute ; pathology ; Proto-Oncogene Proteins c-myc ; drug effects ; metabolism ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo investigate the effect of aminopeptidase N inhibitor ubenimex on differentiation induction of all-trans-retinoic acid (ATRA) in acute promyelocytic leukemia (APL) cells and its mechanism.

METHODSThe expression of CD11b was analyzed by flow cytometry and nitroblue-tetrazolium (NBT) reduction assay was performed to determine the cell differentiation of APL cells. The expressions of c-Myc, ERK1/2, p38MAPK protein and the phosphorylation of ERK1/2, p38MAPK protein in NB4 cells were detected by Western blot assay.

RESULTSUbenimex alone induced no significant changes in NBT reduction activity and CD11b expression but potentiated the differentiation induction activity of ATRA in APL cells. 100 microg/ml of ubenimex could enhance the NBT reduction activity induced by 10 nmol/L of ATRA, intensify the down-regulation of c-Myc protein expression and inhibit the phosphorylation of p38MAPK protein induced by 10 nmol/L of ATRA in NB4 cells.

CONCLUSIONSUbenimex could potentiate ATRA induced differentiation in APL cells, which may be correlated with the inhibition of p38 MAPK protein phosphorylation and regulation of c-Myc protein expression.

Aminopeptidases ; antagonists & inhibitors ; Cell Differentiation ; drug effects ; Drug Synergism ; Flow Cytometry ; Humans ; In Vitro Techniques ; Leucine ; analogs & derivatives ; pharmacology ; Leukemia, Promyelocytic, Acute ; pathology ; Tretinoin ; pharmacology

Objective To investigate the effects of interleukin-18 ( IL-18 ) on M cells of intestinal mucosa in acute pancreatitis (AP) in rats. Methods Seventy SD rats were divided into seven groups: AP 6 hour group, 12 hour group, 24 hour group, IL-18 6 hour group, 12 hour group, 24 hour group, and control group at random. The models of AP were established. Serum, pancreatic tissue, and ileal mucosa were harvested. Serum amylase, glutamate pyruvate transaminse, total bilirubin, endotoxin, IL-27, and TNF-α were detected. Pathologic changes of pancreatic tissue and ileal mucosa were observed. The protein with reverse transcription-polymease chain reaction (RT-PCR). Results The level of serum TNF-α increased obviously in IL-18 6 hour group, 12 hour group, and AP group. The level of serum IL-27 increased in IL-18 6 hour group and AP group and back to normal in IL-18 24 hour group. The in IL-18 group than AP group (P < 0. 05 ). Conclusions IL-18 may affect M cells function and play important roles in intestinal barrier in acute pancreatitis.