This study investigated a nano drug delivery system built by one sort of modified trimethyl chitosan (TMC). The TMC was modified by cRGDyk, ligand of integrin receptor avβ3. Single factor screening was used to optimize the prescription in which the particle sizes of TMC nanoparticle (TMC NPs) and cRGDyk modified TMC nanoparticle (C-TMC NPs) were (240.3 ± 4.2) nm and (259.5 ± 3.3) nm. Electric potential of those two nanoparticles were (33.5 ± 0.8) mV and (25.7 ± 1.6) mV. Encapsulation efficiencies were (76.0 ± 2.2) % and (74.4 ± 2.0) %. Drug loading efficacies were (50.1 ± 2.1) % and (26.1 ± 1.0) %. Then the cellular uptake, uptake mechanism and transport efficacy of TMC NPs and C-TMC NPs were investigated using Caco-2 cell line. The uptake rate and accumulating drug transit dose of C-TMC NPs were 1.98 and 2.84 times higher than TMC NPs, separately. Mechanism investigations revealed that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis were involved in the intercellular uptake of both TMC NPs and C-TMC NPs. What is more, free cRGDyk could remarkably inhibit the uptake of C-TMC NPs.
Biological Transport ; Caco-2 Cells ; Caveolae ; Chitosan ; chemistry ; Clathrin ; Endocytosis ; Humans ; Integrin alphaVbeta3 ; chemistry ; Nanoparticles ; Particle Size ; Pinocytosis

This study investigated a nano drug delivery system built by one sort of modified trimethyl chitosan (TMC). The TMC was modified by cRGDyk, ligand of integrin receptor avβ3. Single factor screening was used to optimize the prescription in which the particle sizes of TMC nanoparticle (TMC NPs) and cRGDyk modified TMC nanoparticle (C-TMC NPs) were (240.3 ± 4.2) nm and (259.5 ± 3.3) nm. Electric potential of those two nanoparticles were (33.5 ± 0.8) mV and (25.7 ± 1.6) mV. Encapsulation efficiencies were (76.0 ± 2.2) % and (74.4 ± 2.0) %. Drug loading efficacies were (50.1 ± 2.1) % and (26.1 ± 1.0) %. Then the cellular uptake, uptake mechanism and transport efficacy of TMC NPs and C-TMC NPs were investigated using Caco-2 cell line. The uptake rate and accumulating drug transit dose of C-TMC NPs were 1.98 and 2.84 times higher than TMC NPs, separately. Mechanism investigations revealed that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis were involved in the intercellular uptake of both TMC NPs and C-TMC NPs. What is more, free cRGDyk could remarkably inhibit the uptake of C-TMC NPs.

Objective To investigate the effects of B lymphocyte-induced maturation protein-1 ( Blimp-1) on the number and function of splenic lymphocytes.Methods The mice with defective Blimp-1 in T cells were generated by cross-breeding B6.Blimp-1flox/flox mice with B6.Lck-Cre mice.The mononuclear lymphocytes isolated from spleen of T cell conditional Blimp-1 knockout (Blimp-1CKO) mice and wild type ( WT) C57/B6 mice were comparatively analyzed.Alterations of CD4+T and CD8+T cell subsets, the secre-tion of cytokines as well as the expression of C-C chemokine receptor type 7 ( CCR7 ) and Sphingosine-1-phosphate receptor 1 (S1P1) in mice from the two groups were analyzed by flow cytometry.The changes of CD19+B cell subsets were also detected.Results Compared with WT mice, the total numbers of mononu-clear cells, T and B lymphocytes were all significantly increased in Blimp-1CKO mice ( P<0.05) .The ab-solute numbers of CD4+T, CD8+T and CD19+CD5+CD1d+B cells in mice form Blimp-1CKO group were higher than those of the control group (P<0.05), however, no significant differences with the percentages of these cell populations were observed between two groups.Higher numbers and percentages of CD19+CD5+B cells were detected in mice from Blimp-1CKO group (P<0.01).The Blimp-1CKO mice showed increased secretion of IFN-γ, TNF-α, IL-17 and IL-2, but decreased expression of CCR7 on CD8+T cells as com-pared with WT mice (P<0.05).No significant differences with the changes of S1P1 were found between the two groups.Conclusion Blimp-1 played an important role in the maintenance of number, phenotype and function of T cells.Furthermore, not only T cells but also B cell subsets in mice were affected by the dele-tion of Blimp-1 in T cells.

OBJECTIVETo investigate the role of human adenovirus type 5 (Ad5) early-region 1A (E1A) in premature senescence of human fibroblasts induced by Ras activation.

METHODSHuman fibroblasts were cotransduced with E1A, E1A1-143, or their vector (BP) and Ha-RasV12 or its vector (WH). The growth curves and percentages of the apoptotic cells were determined.

RESULTSExpression of E1A or Ha-RasV12 in human fibroblasts significantly inhibited the cell growth. Transduction of Ha-RasV12 along with E1A or E1A 1-143 into human fibroblast cells resulted in active and rapid cell proliferation.

CONCLUSIONE1A can rescue human fibroblasts from Ras-induced premature senescence, and the senescence bypassing activity of E1A resides in its NH2 terminus.

Adenovirus E1A Proteins ; genetics ; metabolism ; Apoptosis ; genetics ; physiology ; Cell Transformation, Neoplastic ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; genetics ; physiology ; Fibroblasts ; cytology ; metabolism ; Genes, ras ; physiology ; Humans ; Skin ; cytology ; Transfection

OBJECTIVE: To investigate the expression of matrix metalloproteinase-3 after brain contusion and its applicability for estimating the age of brain contusion. METHODS: Rats had been divided into three groups: control group, sham operation group and brain contusion group. The expression of matrix metalloproteinase-3 at different time was detected by immunohistochemistry and Western blot. RESULTS: By the immunohistochemistry, no staining was observed in control and sham operation groups. The positive staining of MMP-3 appeared 6 hours after contusion, increased gradually in 24 hours and peaked 5 days after contusion, then started to decrease, 14 days after contusion still could be observed. By the Western blot analysis, no expression of MMP-3 was detected in control and sham groups. The positive staining of MMP-3 appeared 6 hours after contusion, increased gradually and maximized 5 days after contusion, then started to decrease, 14 days after contusion still could be found. CONCLUSION Time-order expression of MMP-3 could be used for estimating the age of brain contusion in forensic pathology.
Animals ; Blotting, Western ; Brain Injuries/enzymology* ; Forensic Pathology ; Immunohistochemistry ; Male ; Matrix Metalloproteinase 3/genetics* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors

The purpose of this study was to establish a method for the monitoring of minimal residual disease (MRD) in bone marrow samples of the children with T acute lymphoid leukemia (T-ALL), and to evaluate its value in clinical application. The immuno-phenotype of the leukemic cells were detected by flow cytometry with two sets of 4-color combinations of antibodies against TdT/CD5/cCD3/HLA-DR(+)CD19(+)CD33 and CD34/CD5/cCD3/HLA-DR(+)CD19(+)CD33 in 32 cases of de novo T-ALL and were compared with the results in 10 normal controls. The antibody combination in regions of the two-parameter plots where the leukemic cells appeared were different from the normal cells was screened as the effective combination which was used to monitor MRD in the bone marrows of the T-ALL children after the inductive treatment. The results indicated that the respective effective frequencies of antibodies against TdT/CD5/cCD3/HLA-DR(+)CD19(+)CD33 and CD34/CD5/cCD3/HLA-DR(+)CD19(+)CD33 were 90.6% and 62.5%. 32 cases of childhood T-ALL were successively screened for antibodies combinations of interest and were identified in 100% (32/32) of these cases. After inductive treatment, the positive rate in 129 times of MRD monitoring was 19.4% (25/129) by flow cytometry and 5.43% (7/129) by FAB morphology. It is concluded that monitoring MRD in patients with T-ALL by flow cytometry with two 4 color combinations of fluorescent antibodies is an quick and effective method. The sensitivity of this method is high and it may be of important significance for the treatment and prognostic evaluation in childhood T-ALL.
Adolescent ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Male ; Neoplasm, Residual ; diagnosis ; immunology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology

OBJECTIVETo elucidate the clinical features of patients at early stage of severe acute respiratory syndrome (SARS).

METHODSFifty-three cases of early SARS were studied retrospectively. The data reviewed included those of epidemiology, clinical manifestations, laboratory investigation and roentgenology.

RESULTSThe patients consisted of 24 men and 29 women, aged 10.85 years (mean 38+/-16.7 years), including 9 infected health-care professionals (17.0%). The mean incubation period was 7.3+/-7.0 days (3.14 days). The onset symptoms were characterized by fever (100%), cough (49.1%), maylgia (24.5%), shortness of breath (20.8%), malaise (17.0%),and diarrhea (5.7%). Routine blood test during the first to the fifth day of the disease revealed WBCs less than 4.0x10(9) /L in 33 patients (62.3%), 4.0-10.0x10(9)/L in 18 patients (34.0%), lymphopenia in 36 patients (67.9%), and PLT less than 100.0x10(9) in 7 patients (13.2%). The main abnormal X-ray finding was single (75.4%) or bilateral (15.1%) localized patchy clouding opacity. The decrease of arterial partial pressure of oxygen occurred in 26 patients (49.1%). The damage of several organs was common, including the elevated ALT or AST in 20 patients (37.7%), elevated BUN or SCR in 6 patients (11.3%) and elevated LDH or CK or HBDH in 23 patients (43.4%).

CONCLUSIONThe clinical manifestations of SARS at the early stage were complex. The close monitoring of the blood cell counts, the blood gas analysis and chest radiography might be crucial to the early diagnosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Cell Count ; Blood Gas Analysis ; Child ; Early Diagnosis ; Female ; Humans ; Male ; Middle Aged ; Radiography, Thoracic ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; diagnosis ; epidemiology

OBJECTIVETo develope a tree analysis pattern of mass spectral urine profiles to discriminate bladder transitional cell carcinoma (TCC) from non-cancer lesions using surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technology.

METHODSUrine samples from 61 bladder transitional cell carcinoma (TCCs) patients, 53 healthy volunteers and 42 patients with other urogenital diseases were analyzed using IMAC-Cu-3 ProteinChip. Proteomic spectra were generated by SELDI-TOF- MS. A preliminary "training" set of spectra derived from analysis of urine from 46 TCC patients, 32 patients with benign urogenital diseases (BUD), and 40 age-matched unaffected healthy men were used to train and develop a decision tree classification algorithm which identified a fine-protein mass pattern that discriminated cancers from non-cancers effectively. A blinded test set including 38 cases was used to determine the sensitivity and specificity of the classification system.

RESULTSThe algorithm identified a cluster pattern that, in the training set, segregated cancer from non-cancer with a sensitivity of 84.8% and specificity of 91.7%. The discriminatory pattern was correctly identified. A sensitivity of 93.3% and a specificity of 87% for the blinded test were obtained when compared the TCC versus non-cancers.

CONCLUSIONSELDI-TOF-MS technology is a rapid, convenient and high-throughput analyzing method. The urine tree analysis proteomic pattern as a screening tool is effective for differential diagnosis of bladder cancer. More detailed studies are needed to further evaluate the clinical value of this pattern.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Transitional Cell ; diagnosis ; urine ; Cystitis ; diagnosis ; urine ; Decision Trees ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; diagnosis ; urine ; Protein Array Analysis ; Proteomics ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Urinary Bladder Neoplasms ; diagnosis ; urine

OBJECTIVETo evaluate the bioequivalence of tiopronin enteric capsules (testing preparation, T) versus tablets (reference preparation, R).

METHODSA single oral dose of tiopronin enteric capsules or tablets at 200 mg was administered in 2 groups of Chinese healthy volunteers (n=9) in a randomized crossover design at the interval of 2 weeks. The plasma concentrations of tiopronin were measured by HPLC-MS/MS, and the pharmacokinetic parameters were calculated by DAS 2.0 program. The bioequivalence between the two preparations was evaluated.

RESULTSThe main pharmacokinetic parameters were as follows: C(max)(microg.ml(-1)) 3.612-/+1.2393 (R), 3.644-/+1.540 (T); t(max) 4.333-/+1.0853 (R), 3.611-/+1.420 (T); t((1/2))(h) 18.245-/+11.270 (R), 23.403-/+10.500 (T); AUC0-t (microg.h.ml(-1)) 18.732-/+6.92318 (R), 18.713-/+6.585 (T); AUC0-infinity (microg.h.ml(-1)) 21.900-/+7.31220 (R), 20.780-/+7.965 (T). The relative bioavailability of tiopronin enteric capsule was 103.712-/+23.956%, with 90% confidential intervals of ln(AUC0-->72), ln(AUC0-infinity) and ln(C(max)) of 91.1%-111.8%, 96.8%-118.3%, and 85.1%-113.0%, respectively.

CONCLUSIONThe tiopronin enteric capsules were bioequivalent to the tablets.

Biological Availability ; Capsules ; Cross-Over Studies ; Health ; Humans ; Linear Models ; Male ; Reproducibility of Results ; Therapeutic Equivalency ; Tiopronin ; pharmacokinetics ; Young Adult

The research was aimed to detect the expression levels of retinoblastoma protein (pRb) in child acute leukemia cells, and to explore its possible association with leukemia cells cycle, the risk of disease, minimal residual disease (MRD) monitoring and prognosis of B-ALL. Flow cytometry (FCM) was used to detect the expression of pRb in 89 cases of acute leukemia (including 25 AML, 10 T-ALL and 54 B-ALL) and bone marrows from 7 normal children (control group). Meanwhile the cell cycle in some cases was analyzed. The results showed that (1) the FCM could accurately detect the expression of pRb in acute leukemia cells; (2) the high level of pRb expression was frequent in all types of child acute leukemias. In the same case, the expression of pRb was significantly increased in leukemia cells when compared with non-leukemia cells. And no detectable pRb protein was found in partial cases of acute leukemia; (3) there was a close relation between expression of pRb and the cell cycle of leukemia cells, the number of G(1) phase cells in pRb positive case of B-ALL was more than that in pRb negative case (92% vs 77%); (4) in B-ALL, the level of pRb expression in MRD positive group was significantly lower than that in MRD negative group (P < 0.05), but pRb expression was stable in non-leukemia cells during therapy; (5) pRb expression was related to the early response to therapy in B-ALL, the expression of pRb was significantly increased in sensitive group when compared with insensitive group (P < 0.05). It is concluded that high level or absence of pRb expression can be found in child acute leukemia cells. The expression of pRb is positively related to cell cycle of leukemia cells, MRD monitoring and the early response to therapy. In short, the detection of pRb expression level can guide the therapy and the evaluation of prognosis in B-ALL.
Burkitt Lymphoma ; metabolism ; Child ; Female ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; Male ; Neoplasm, Residual ; Prognosis ; Retinoblastoma Protein ; biosynthesis ; genetics