Objective To explore the effect of surgical treatment on primary gastrointestinal non-Hodgking lymphoma(NHL) in children.Methods Nine cases of clinical and follow-up data of primary gastrointestinal NHL were studied retrospectively to evaluate the effect of surgical treatment on primary gastrointestinal NHL in children.Results Seven cases were male and 2 cases were female.The mean age was(5.59?3.27)years old.The clinical manifestation included abdominal mass (7 cases),abdominal pain (5 cases),fever (2 cases),haematemesis and melena (2 cases),constipation (1 case) and paroxysmal abdominal pain with vomiting (1 case).Nine cases were diagnosed as primary gastrointestinal NHL,including 1 case of intussusception,1 case of acute appendicitis,2 cases of gastrointestinal obstruction,2 cases of gastrointestinal bleeding and 3 cases of abdominal mass.One case received the operation of intussusception reduction,tumor resection and intestinal anastomosis.One case received appendectomy.One case received the operation of tumor biopsy and transverse colon colostomy.Six cases received laparotomy.Six cases were diagnosed as Burkitt lymphoma.One case was anaplastic large cell lymphoma and 2 cases were diffuse large B-cell lymphoma.One case was at stage Ⅰ,1 case was at stage ⅠE,2 cases were at stage Ⅱ,3 cases were at stage ⅡE and 2 cases were at stage Ⅲ.Nine patients had received operation.One case died after operation and 8 cases had received combined chemotherapy.The 1 and 3 years survival rates were 75.0% and 37.5%,respectively.Conclusions Acute abdomen is often the first symptom of primary gastrointestinal NHL in children and comprehensive surgical treatment is an effective procedure for it.

OBJECTIVETo observe the effect of sensory neuropeptide substance P combined with epidermal stem cells (ESC) on wound healing and nerve regeneration in diabetic rats.

METHODSESC that had been isolated from SD rats were identified and cultured in vitro, and they were inoculated onto nourishing layer of amniotic membrane to construct amniotic membrane-ESC. Four full-thickness skin wounds were produced on the back of each of 48 diabetic rats. The resulted 192 wounds were randomly divided into ESC + substance P group, ESC group, substance P group, and control group according to the lottery method, with 48 wounds in each group. Wounds in ESC + substance P group and ESC group were transplanted with amniotic membrane-ESC, and those in substance P group and control group were transplanted with amniotic membrane. After transplantation, 250 µL substance P in the concentration of 1 × 10(-7) mol/L was injected around and into the middle of the wounds in ESC + substance P group and substance P group, 2 times a day, and continued for 4 days, while 250 µL PBS solution was injected in the above-mentioned position in ESC group and control group as control, 2 times a day, and continued for 4 days. On post injury day (PID) 4, 7, 10, 14, 17, and 23, the wound healing rate (with 8 wounds at each time point) was observed and determined, and changes in wound tissue structure were observed with HE staining. On PID 4, 7, and 10, collagen distribution in wound tissue was observed with Masson staining, and type I and type III collagen deposition in wound tissue was respectively observed after immunohistochemical staining. The distribution of protein gene product 9.5 (PGP 9.5) and regeneration of substance P positive nerve fibers in wound tissue were observed with immunohistochemical staining on PID 14 and 23. Data were processed with one-way analysis of variance and t test.

RESULTS(1) The wound healing rate in ESC + substance P group reached 100.0% on PID 14, which was obviously earlier than that in ESC group, substance P group, and control group, healing was respectively observed on PID 17, 17, and 23. The wound healing quality in ESC + substance P group was better than that in the other three groups as shown by HE staining. (2) On PID 10, collagen that was darkly stained and widely distributed was observed in wound tissue of ESC + substance P group and substance P group, while collagen in the other two groups was lightly stained and narrowly distributed. Deposition quantity of type I collagen gradually increased, and that of type III collagen gradually decreased in the wounds of each group over time. On PID 4, 7, and 10, distribution amount of type I collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.72, 118.21, 26.71, P values all below 0.01) and control group (with t value respectively 44.37, 22.76, 30.32, P values all below 0.01), while there was no significance between ESC + substance P group and substance P group. On PID 4, 7, and 10, distribution amount of type III collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.27, 28.68, 14.51, P values all below 0.01) and control group (with t value respectively 35.68, 22.52, 22.24, P values all below 0.01). (3) A large amount of PGP 9.5 and regeneration of substance P positive nerve fibers, and some peripheral nerve fibers in deep wound extending to epidermis were observed in wound tissue of ESC + substance P group and substance P group. A small amount of PGP 9.5 and regeneration of substance P positive nerve fibers without peripheral nerve fibers extending to epidermis were observed in deep wound tissue of ESC group and control group. On PID 14, 23, ratios of area of PGP 9.5 positive nerve fiber in the wounds of ESC + substance P group were (3.86 ± 0.25)% and (7.03 ± 0.28)%, and they were significantly higher than those of ESC group [(1.48 ± 0.30)%, (3.01 ± 0.43)%, with t value respectively 23.95, 30.27, P values all below 0.01] and control group [(1.46 ± 0.23)%, (2.84 ± 0.29)%, with t value respectively 27.35, 40.32, P values all below 0.01]. On PID 14, 23, ratios of substance P positive nerve fiber area in the wounds of ESC + substance P group were (2.01 ± 0.14)% and (1.19 ± 0.11)%, which were obviously higher than those of ESC group [(0.85 ± 0.17)%, (1.34 ± 0.21)%, with t value respectively 20.50, 2.60, P < 0.05 or P < 0.01] and control group [(0.74 ± 0.15)%, (1.30 ± 0.17)%, with t value respectively 23.98, 2.41, P < 0.05 or P < 0.01].

CONCLUSIONSJoint application of substance P and ESC can effectively promote healing of wound and nerve regeneration in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Epithelial Cells ; cytology ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; Substance P ; pharmacology ; therapeutic use ; Wound Healing

OBJECTIVE To establish a RP-HPLC method for determination of simvastatin and its related substance lovastatin. METHODS The chromatographic conditions were: a Waters Symmetry C18 column (250 mm×4.6 mm, 5 μm), a mixture of acetonitrile-sodium dihydrogen phosphate (pH 5.4) (65∶35) as mobile phase, flow rate 1.0 mL·min-1, and detected at 238 nm. RESULTS The linear ranges of lovastatin and simvastatin were 0.3～3.0 μg·mL-1,0.03～0.30 mg·mL-1, respectively. The average recovery were 100.2% (RSD=1.5%) and 99.4% (RSD=1.7%), respectively. CONCLUSION The method is simple, quick, sensitive, accurate, and reproducible. It can be used to the quality control of synthetic simvastatin products.

To observe the expressions of CD10 in childhood B-acute lymphoblastic leukemia (B-ALL) and to define the role of CD10 in minimal residual disease (MRD) detection. 58 cases of childhood B-ALL were studied in this program. Four-color flow cytometry was used to analyze the characteristics of B-ALL phenotypes. The four-color fluorochrome labeled antibody combinations of CD10 with other markers were used to detect MRD. The results showed that CD10 overexpression (CD10(bright)) was detected in 65.5% (38/58) of B-ALL patients and a strong correlation between CD10(bright) and CD34 expression was also observed, i.e. CD10(bright) expression most frequently happened in B-ALL with high percentage of CD34 positive cells. In detection of MRD, CD10(bright), combined with other markers, could effectively distinguish normal cells with leukemic cells, even if there was no any other marker that can be used. It is concluded that CD10(bright) expression correlates with high expression of CD34 in B-ALL, it is a good marker for MRD detection. The combination of CD10 and other markers can be applied in B-ALL MRD detection with flow cytometry.
Adolescent ; Antigens, CD34 ; analysis ; Burkitt Lymphoma ; diagnosis ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Male ; Neoplasm, Residual ; Neprilysin ; analysis ; physiology

OBJECTIVETo study the in vitro antitumor activity of bcl-2 fully phosporothioated antisense oligodeoxynucleotide (bcl-2 ASODN) to malignant lymphoblastic cells.

METHODSProliferation and apoptosis of Raji cells incubated with bcl-2 ASODN were evaluated by MTT assay, flow cytometry (FCM) and electron microscopy, and the level of bcl-2 protein and mRNA expression were assessed by FCM and RT-PCR, respectively.

RESULTSMTT assay demonstrated that bcl-2 ASODN could partially inhibit the growth of Raji cells. After incubated with ASODN for 48 hours, Raji cells exhibited characteristic morphologic changes of apoptosis, including cytoplasm membrane blebbing, chromatin condensation crescents formation and nuclear fragmentation. The apoptosis rate of Raji cells treated with 20 micromol/L bcl-2 ASON for 72 hrs was 43.86% which is significantly higher than that of control (10.05%). The bcl-2 ASODN induced apoptosis of Raji cells was accompanied by declined expression of bcl-2 mRNA, which decreased to 0.88% at 72 hrs and was significantly lower than that of control (79.54%).

CONCLUSIONbcl-2 ASODN induced Raji cells apoptosis by downregulating bcl-2 protein.

Apoptosis ; drug effects ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured ; drug effects ; metabolism

This study was aimed to investigate the effect of recombinant mutant human TNF-related apoptosis-inducing ligand (rmhTRAIL) combined with As(2)O(3) on inducing apoptosis of adriamycin-resistant leukemia cell line K562/A02 (mdr-1(+)). The morphologic changes of cells treated with rmhTRAIL were observed by inverted microscope, taking adriamycin-sensitive cell line K562 (mdr-1(-)) as control; the inhibitory rate of cell proliferation after being treated with rmhTRAIL, As(2)O(3) alone or combined was assayed by MTT method; the apoptosis peaks of K562/AO2 and K562 were quantitatively detected by flow cytometry with PI staining after being treated with rmhTRAIL, As(2)O(3) alone or in combination. The results indicated that the inhibition effect of rmhTRAIL and As(2)O(3) in combination on K562/AO2 and K562 cells was higher than that of riTRAIL and As(2)O(3) alone (p < 0.01), rmhTRAIL combined with As(2)O(3) had synergistic effect in killing K562/AO2 and K562 cells by king's formula. The apoptosis rates of K562/AO2 and K562 cells were 34.93 +/- 0.10% and 10.53 +/- 0.16% (p < 0.01), as well as 5.95 +/- 0.07%, and 3.50 +/- 0.01% (p < 0.05), 50.95 +/- 0.91% and 20.75 +/- 0.95% (p < 0.05) respectively when their cells were treated by rmhTRAIL and As(2)O(3) alone. The apoptosis rate in K562/AO2 group was higher than that in K562 group. It is concluded that rmhTRAIL can induce K562/A02 and K562 cell apoptosis; rmhTRAIL combined As(2)O(3) had synergistic effects; the efficacy of on rmhTRAIL or As(2)O(3) inducing K562/AO2 cell apoptosis is higher than that on their parental cell line K562.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Oxides ; pharmacology ; Recombinant Proteins ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

OBJECTIVETo probe into the occurrence rates of the effective antigen combinations which were used to detect the minimal residual disease (MRD) by flow cytometry in childhood B-lineage acute lymphoblastic leukemia (B-ALL), as well as the relationship between clinical-biologic factors and different combinations.

METHODSAmong the 327 B-ALL children enrolled in our study, 289 cases were identified with at least one antigen combination as MRD marker. Their bone marrow samples were monitored by using 9 combinations with 4 antigens each, analyzed the occurrence rates and compared them with international reports. Also the differences in the distribution of each antibody combination in the different clinical-biologic groups were compared by the chi-square test and Fisher's exact test.

RESULTS(1) Totally 327 cases of childhood B-ALL were screened for antibody combinations of interest and 88.4 percent of them (289 cases) were identified with effective antibody combinations. (2) The occurrence frequencies of antigen combinations were different. The highest frequency was seen with TdT/CD10/CD34/CD19 combination which was 70.59 percent. Expressions of antigen combinations in Chinese children were different from those in western countries. (3) Some antibody combinations presented different frequency among different clinical groups. CD38/CD10/CD34/CD19 was expressed more often in samples of relapsed patients (P = 0.045). CD66c/CD10/CD34/CD19 expression was significantly higher in BCR/ABL positive group (P = 0.037) and relapsed patients group (P = 0.047). TdT/CD10/CD34/CD19 was expressed more in MLL-AF4 negative group (P = 0.005) and Prednisone Good Response group (P = 0.002). CD58/CD10/CD34/CD19 was correlated with low relapse rate (P = 0.032).

CONCLUSION(1) The coverage rate of 9 antigen combinations in our study was 88.4%. The occurrences of frequency of different antibody combinations in B-ALL were different, and also different from that of western countries. The occurrence frequencies of antibody combinations CD21/CD10/CD34/CD19, CD22/CD10/CD34/CD19, CD10/CD56/CD34/-CD19 and TdT/Cu/CD34/CD19 were lower than those of the western report, while CD38/CD10/-CD34/CD19, CD45/CD19/CD10/CD34, CD58/CD10/CD34/CD19 and CD66c/CD10/CD34/CD19 were similar to those of the reports from western countries. (2) TdT/CD10/CD34/CD19 may work as a simplified method to detect MRD in Chinese population. (3) The occurrence frequency of CD38/CD10/CD34/CD19, CD45/CD19/CD10/CD34, CD58/CD10/CD34/CD19, TdT/CD10/CD34/CD19 could be effective remediation and evidence to evaluate the remission quality and guide the therapy, especially for those with no original MRD marker record. (4) CD58/CD10/CD34/CD19 and TdT/CD10/CD34/CD19 may correlate with good prognosis, but CD66c/CD10/CD34/CD19 and CD38/CD10/CD34/CD19 may predict poor prognosis. These results might contribute to individual risk evaluation and guide the therapy selection.

Child ; Child, Preschool ; Humans ; Infant ; Leukemia, B-Cell ; immunology ; therapy ; Neoplasm, Residual ; Neprilysin ; immunology ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; therapy

The present study was purposed to investigate the inhibition effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on growth of RPMI8226 cells and adhesion between RPMI8226 cells and bone marrow stroma cells (BMSC), and to explore its mechanism as well. The inhibition effects of TRAIL on cells growth and adhesion were assayed by MTT; cell apoptosis was detected by Annexin V and PI; expression of genes bax, bcl-2, mcl-1, CARP1, CARP2, XIAP and cFLIP were determined by semi-quantitative RT-PCR; apoptosis-related protein expression was detected by Western blot. The results showed that TRAIL inhibited the proliferation of RPMI8226 cells in dose- and time-dependent manners. TRAIL induced apoptosis in RPMI8226 cells, the expression level of genes bcl-2, mcl-1, CARP1, CARP2, XIAP and cFLIP decreased, while the expression level of Bax increased, but the expression level of caspase-3 and NF-kappaB P65(RelA) proteins decreased. Moreover, TRAIL up-regulated the expression level of adherent molecule CXCR4 in RPMI8226 cells significantly. It is concluded that TRAIL up-regulated the expression level of adherent molecule CXCR4 in RPMI8226 cells significantly, and induced the apoptosis of RPMI8226 cells. Growth inhibition effect of TRAIL on RPMI8226 cells is in dose- and time-dependent manners.
Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; metabolism ; Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Multiple Myeloma ; metabolism ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; pharmacology

In order to investigate the role and the mechanism of protein tyrosine phosphatase (PTPase) signaling pathway in the regulation of proliferation, cell cycle and apoptosis in lymphoma cells, the effects of sodium orthovanadate, Na(3)VO(4), a specific PTPase inhibitor, were explored on Raji lymphoblast-like cell line by MTT assay and CFU-Raji culture, morphologic observation, DNA gel electrophoresis, FCM and RT-PCR. Results showed that MTT assay and CFU-Raji culture demonstrated that sodium or thovanadate inhibited the growth of Raji cells in a concentration-dependent fashion; morphologic observations showed that Raji cells exhibited cytoplasm shrinkage, cytoplasm membrane blebbing, nuclear fragmentation and chromatin condensation forming crescents along nuclear membrane characteristic of apoptosis in the presence of Na(3)VO(4); DNA gel electrophoresis revealed typical DNA ladder reminiscent of DNA cleavage at internucleosomal sites in Na(3)VO(4) treated cells; FCM and RT-PCR indicated that Na(3)VO(4) intervention increased the fraction of annexin V(+) PI(-) cells, reduced the value of mitochondrial transmembrane potential, induced G(2)/M arrest and down-regulated the expression of Bcl-2 and cyclin B1 at both mRNA and protein level in a concentration-dependent manner. It was concluded that PTPase pathway might be implicated in the regulation of cell proliferation, cell cycle and apoptosis, and PTPase specific inhibitor Na(3)VO(4) could induce Raji cell growth inhibition, G(2)/M arrest and apoptosis via down-regulation of Bcl-2 and cyclin B1, and reduction of mitochondrial transmembrane potential.
Apoptosis ; drug effects ; Cell Division ; drug effects ; Cyclin B ; analysis ; Cyclin B1 ; Enzyme Inhibitors ; pharmacology ; Humans ; Leukocyte Common Antigens ; analysis ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; physiology ; Protein Tyrosine Phosphatases ; antagonists & inhibitors ; Vanadates ; pharmacology

In order to investigate the anti-tumor activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors and the mechanism underlying the cell proliferation and apoptosis modulated in myeloma cells, the effects of mevastatin, an HMG-CoA reductase inhibitor, on cell growth, cell cycle progression and apoptosis in U266 human multiple myeloma (MM) cell line in vitro were explored by MTT colorimetric assay, morphologic observation, flow cytometry, DNA gel electrophoresis, and RT-PCR. The results demonstrated that mevastatin inhibited the growth of U266 cells in time- and dose-dependent manners. Cell cycle analysis showed that U266 cells underwent G(0)/G(1) arrest under exposure to mevastatin, but it did not affect p27 expression at both mRNA and protein level. Morphologic observations revealed cytoplasm shrinkage, nuclear condensation and fragmentation in mevastatin-treated cells, and fraction of annexin V(+)PI(-) cells increased significantly in the presence of the agent as determined by flow cytometric assay. In addition, mevastatin caused the collapse of mitochondrial transmembrane potential (Deltapsim), induced DNA fragmentation, and down-regulated the mRNA expression of bcl-2. The growth-inhibitory, cell cycle arresting, and proapoptotic effects of mevastatin in U266 cells could be effectively reversed by the addition of mevalonate (MVA), the immediate endproduct of the reaction catalyzed by HMG-CoA reductase. It is concluded that mevastatin suppresses proliferation by inducing G(0)/G(1) phase arrest and triggering apoptosis via down-regulation of bcl-2 and reduction of Deltapsim, which may be attributed to the inhibition of MVA pathway by mevastatin. Statins including mevastatin may find their future application in the treatment of MM.
Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; G1 Phase ; drug effects ; Genes, bcl-2 ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Lovastatin ; analogs & derivatives ; pharmacology ; Multiple Myeloma ; drug therapy ; pathology