OBJECTIVETo study the inhibitory effects of matrine, in different concentrations, on invasion and metastasis of human acute lymphocytic leukemia cell line Jurkat.

METHODSIn vitro cultured Jurkat cells were treated by matrine in concentration of 0 g/L, 0.1 g/L, 0.15 g/L and 0.2 g/L, respectively. Then cell adhesion assay, cell migration assay and matrigel invasion assay were used respectively to observe the effects of matrine on adhesion, migration and invasive capacity of Jurkat cells. Meantime, RT-PCR was performed to detect the matrix metalloproteinase (MMP)-2 and MMP-9 mRNA expression levels. Comparison of measurement data among groups was analyzed by variance analysis.

RESULTSAs compared with the control group, the adhesion of Jurkat to fibronectin (FN) was significantly inhibited by 0.15 g/L and 0.2 g/L of matrine (P < 0.05); the cell migration and invasive capacity were significantly lowered by 0.1 g/L, 0.15 g/L and 0.2 g/L matrine (P < 0.01). High mRNA expression of MMP-9 presented but that of MMP-2 was expressed insignificantly in Jurkat cells, matrine at 0.1 g/L, 0.15 g/L and 0.2 g/L showed obvious effect in down-regulating MMP-9 mRNA expression (P < 0.01). Besides, MMP-9 mRNA expression was found to be positively correlated with the invasive capacity of Jurkat cells (r = 0.940, P < 0.01).

CONCLUSIONSMatrine is a good drug for antagonizing the invasion and metastasis of leukemia cells, it may roundly inhibit the adhesion, migration and invasive capacity of Jurkat cells, the mechanism might be related with the down-regulation of MMP-9 mRNA expression.

Alkaloids ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Jurkat Cells ; Leukemia ; drug therapy ; pathology ; physiopathology ; Mice ; NIH 3T3 Cells ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Quinolizines ; pharmacology ; Random Allocation

OBJECTIVETo study the potential effect of hypoxia on invasion and metastasis of leukemia cell line K562.

METHODSK562 cells were cultured with the conventional method in vitro and treated with 1%, 3% and 5% oxygen for 24 hrs. The normoxic cultured K562 cells were used as the control group. Cell adhesion assay, cell migration assay and cell invasion assay were used to detect the adhesion, migration and invasion abilities of K562 cells. RT-PCR was used to measure the mRNA expression of HIF-1alpha, VEGF, MMP-2 and MMP-9. The protein level of HIF-1alpha was measured by Western blot.

RESULTSCompared with the control group, the 3% and 5% oxygen treatment groups significantly increased the adhesion, migration and invasion abilities of K562 cells (p<0.05 or <0.01), and up-regulated the protein level of HIF-1alpha and the mRNA levels of HIF-1alpha,VEGF, MMP-9 and MMP-2 (p<0.05 or 0.01). However, there were no significant differences in the above indexes between the 1% oxygen treatment and the control groups.

CONCLUSIONSModerate hypoxia can enhance the abilities of invasion and metastasis of K562 cells, probably by an up-regulation of HIF-1alpha level and VEGF, MMP-2 and MMP-9 mRNA expression.

Animals ; Cell Adhesion ; Cell Hypoxia ; Cell Movement ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; K562 Cells ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; Mice ; NIH 3T3 Cells ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo investigate the clinical features of nutritional iron deficiency anemia (IDA) and analyze the risk factors for the severity of anemia, and to provide a basis for the prevention and clinical diagnosis and treatment of this disease.

METHODSA retrospective analysis was performed on the clinical data of 372 children with IDA to investigate the risk factors for the severity of IDA.

RESULTSOf 372 cases, the male-to-female ratio of these patients was 2.72 : 1. Of all cases, 79.9% were aged 6 months to 2 years, and 30.7% were premature infants; 22.9% had a birth weight of < 2.5 kg, and 77.1% had a birth weight of ≥2.5 kg; 36.0% were delivered by natural birth, and 64.0% were delivered by caesarean section; 79.3% were not given solid foods in time; 46.2% had a history of lower respiratory tract infection/recurrent upper respiratory tract infection, diarrhea, trauma, or surgery. The univariate analysis showed that age, birth weight, gestational age, timely introduction of solid foods, and a history of lower respiratory tract infection/recurrent upper respiratory tract infection, diarrhea, trauma, or surgery were associated with the severity of anemia. The multivariate analysis showed that birth weight and the mentioned medical history were associated with the severity of anemia.

CONCLUSIONSNutritional IDA is common in children aged 6 months to 2 years. Nowadays, improper feeding pattern is still one of the main causes of IDA. Birth weight and a history of lower respiratory tract infection/recurrent upper respiratory tract infection, diarrhea, trauma, or surgery are closely associated with the severity of anemia.

Adolescent ; Anemia, Iron-Deficiency ; etiology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Logistic Models ; Male ; Retrospective Studies ; Risk Factors

Objective To study the clinical characteristics of bulbar myasthenia gravis (MG). Methods Retrospective review was performed on 166 patients with bulbar type of myasthenia gravis, diagnosed at Tongji Hospital in the period of May 1983 through October 2005.Results Bulbar MG was a relatively rare type of MG,accounting for 5.7% (166/2888) of MG classifications.Females were more often affected than males (the ratio of male:female was 1:1.35).The peak of onset age was at 20—40 years.The incidence of myasthenia crisis in the group was 26.5% (44/166).Myasthenic crisis occurred in 10.8% (18/166) of the bulbar MG patients within 6 months after onset,resulting in a mortality rate of 6.0% (10/166) in the group.Out of the group,30 cases experienced puhnonary infections (18.1%). Thirty cases were initially misdiagnosed as other diseases such as nasopharyngeal disorders (33/166, 19.9%).The routine therapy was not very satisfactory.Median dose cyclophosphamide therapy appeared to be effective for ameliorating refractory MG.Thymectomy was performed in 25 patients,with optimistic efficacy rate up to 80.0% (20/25) in a 3-year follow-up.Conclusions The clinical analysis in the current study suggested that the bulbar MG had its own characteristics in such aspects as progression of the disease, complications,treatment and prognosis.The information of the clinical manifestations presented in this study may be useful in diagnosing and treating bulbar MG.

BACKGROUNDThe technique of intersphincteric resection of tumors combined with coloanal anastomosis has been used to avoid permanent colostomy for patients with a rectal cancer located < 5 cm from the anal verge. This study aimed at assessing the preservation of continence function of the residual rectum and the clinical prognosis of patients with lower rectal cancer after intersphincteric resection using a prolapsing technique.

METHODSThis study included patients with the following inclusion criteria: (1) pathological evidence of rectal cancer and the tumors within distal margins located 5 cm or less from the anus by preoperative endoscopic examination; (2) no evidence by MRI of infiltration of either the external sphincter, puborectalis or the levator muscle; (3) the patients are eligible for intersphincteric resection and lower coloanal anastomosis with a preoperative biopsy showing the tumors with well-to-moderate differentiation. From January 2000 to June 2004, 23 patients with low rectal cancer were included in this study. We used the standard abdominoperineal approach to perform radical resection of tumors with excision of the mesorectum and total or part of the internal sphincters. The patients were followed for assessment of the function of the residual rectum and of cancer recurrence after the operations.

RESULTSThe median tumor distance from the anal margin was 4.5 (range 3.5 - 5.0) cm and the mean distal surgical margin 1.6 (range 1.0 - 2.0) cm. Cancer was classified into Stage I (30.4%), Stage II (47.8%), and Stage III (21.7%) according to the TNM classification. Two patients developed anastomotic fistula after the surgical resection and 2 patients (8.7%) developed later stages of anastomotic stricture at the site of coloanal anastomosis. The median follow-up period was 31.5 months (range 12 - 54) and 2 patients (8.7%) developed local recurrence. Three deaths were associated with distal organ metastasis. Twenty patients (87.0%) have maintained competence to control solid or liquid stool and the capacity of flatus continence after the surgery. Among these patients, 2 patients were able to control solid stool and occasionally lose continence of liquid stool. And only 1 patient (4.4%) has retained partial rectum function with good continence of solid stool but not liquid after the operations. Average times of defecation per day of 3, 6, 12, 24 and 36 months after the surgery were 13.1, 4.7, 3.1, 2.9, and 3.2 times/day. Anal manometer measurements showed a decrease of pressure during the resting time after intersphincteric resection and this change remained during the period of follow-up. The maximum squeeze pressure was improved after an initial decrease after the surgery.

CONCLUSIONSMore residual rectum function after the surgery may be preserved by intersphincteric resection of low rectum cancer. At the same time this technique is safe with few postoperative complication and low tumor recurrence after the surgery.

Digestive System Surgical Procedures ; methods ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; physiopathology ; surgery ; Rectum ; pathology

OBJECTIVETo investigate the clinical features of Epstein-Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH), to analysis the outcome of HLH-2004 protocol, and to explore the prognostic factors in EBV-HLH patients.

METHODSThe clinical features at onset and outcome of HLH-2004 protocol from 83 pediatric patients with EBV-HLH enrolled from January 2006 to December 2009 in our hospital were analyzed retrospectively. Univariate and multivariate COX regression analysis were used to identify statistically significant prognostic factors.

RESULTS(1) Among the 83 patients, 45 were males and 38 were females. The age of onset ranged from 6 months to 14 years 4 months. 44 patients were treated with HLH-2004, and 3-year overall survival (OS) was (55.8 ± 7.9)%. (2) The most common clinical features of EBV-HLH included high fever, cytopenia, hepatosplenomegaly, and coagulopathy; The respiratory symptoms, angina phlogistic, skin rashes, neurologic abnormality were rare. 97.3% of patients showed an elevation of serum ferritin, liver dysfunction and lipid metabolism disorders was found in most of EBV-HLH patients. 89.0% of patient had hemophagocytosis in bone marrow at diagnosis of EBV-HLH. (3) COX regression analysis revealed that anemia degree, serum albumin < 30 g/L, CD4:CD8 abnormity, NK cell < 3%, treatment protocol were related with the prognosis significantly (P < 0.05).

CONCLUSIONEBV-HLH in pediatric patients has severe clinical feature and poor prognosis. HLH-2004 protocol is an effective treatment for patients with EBV-HLH. Symptomatic treatment can't rescue the patients of EBV-HLH.

Adolescent ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; drug therapy ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Killer Cells, Natural ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; virology ; Male ; Prognosis ; Retrospective Studies ; Treatment Outcome

OBJECTIVEThe abnormality of hemopoietic inductive microenvironment (HIM) is involved in the pathophysiology of aplastic anemia (AA). Mesenchymal stem cells (MSC) are main source of bone marrow stromal cells which constitute the bone marrow HIM. Thus, the bone marrow failure in AA may be related to the function of MSC. The aim of the study was to investigate the hematopoiesis support function of MSC in children with AA in vitro.

METHODSBone marrow samples were collected from 24 children with AA at diagnosis and 19 children with idiopathic thrombocytopenic purpura (ITP), infectious mononucleosis or lymphadenitis (controls). MSCs from bone marrow samples were isolated, cultured and expanded. Morphology, proliferation activity and colony forming unit-fibroblast (CFU-F) were measured. The ability of bone marrow MSC to adhere hemopoietic cells was assayed by MTT. The concentration of stem cell factor (SCF) released from MSC was tested using ELISA. Mononuclear cells (MNC) of bone marrow were plated onto a feeder layer formed by MSC. Cells count and BFU-E, CFU-GM, CFU-GMME productions were measured.

RESULTSThe first and third passage time of MSC in children with AA was longer than that in the controls. The number of CFU-F in children with AA (15.70+/-5.78) was less than that in the controls (21.73+/-5.74) (P<0.05). The concentration of SCF in MSC supernatants in children with AA (30.69+/-16.82 pg/mL) was significantly lower than the controls (50.74+/-14.83 pg/mL) (P<0.01). The total MNC count and the number of BFU-E, CFU-GM and CFU-GMME colonies in the support of MSC in children with AA were significantly lower than those in the controls (P<0.01).

CONCLUSIONSThe hematopoiesis support function of MSC was significantly reduced in children with AA in vitro. The decreased hematopoiesis support function of MSC may be related its decreased proliferation capacity and SCF release activity.

Adolescent ; Anemia, Aplastic ; physiopathology ; Cell Adhesion ; Child ; Child, Preschool ; Female ; Hematopoiesis ; Humans ; Leukocytes, Mononuclear ; physiology ; Male ; Mesenchymal Stromal Cells ; physiology ; Stem Cell Factor ; physiology

OBJECTIVETo summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment.

METHODThe process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed.

RESULTSThe main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding.

CONCLUSIONFor children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.

Blood Coagulation Disorders ; chemically induced ; diagnosis ; therapy ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Rodenticides ; poisoning ; Vitamin K 1 ; administration & dosage ; therapeutic use

OBJECTIVETo analyze the clinical and laboratory data from acute lymphoblastic leukemia (ALL) patients and the results of treatment using 04 Protocol (suggested by the Pediatric Hematology Group of Chinese Medical Association in 2004).

METHODSThis study included 88 children with ALL below the age of 18 years during the period from October 1, 2004 to June 30, 2007. Minimal inhibitory concentration (MIC) and clinical risk classification were done and the new chemotherapy regimen was used according to the protocol. Patients were stratified into low-risk (LR), medium-risk (MR), and high-risk (HR) groups. Life table method was used to estimate survival rate and statistical analysis was done by using software SPSS for Windows.

RESULTSFrom October 2004 to June 2007, 88 childhood ALL patients were treated with the 04 Protocol. Sixty-three (91.30%) patients attained complete remission (CR) and 17 patients lost to follow up. The overall 4-year-event-free survival (EFS) rate (+/- SE) was (59.73 +/- 7.22)%. EFS was (75.60 +/- 9.71)% in the LR (n = 30), (65.50 +/- 11.69)% in the MR (n = 20) and (44.03 +/- 12.36)% in the HR. Relapse occurred in 18.18% of patients. Seven (7.95%) of 88 patients with ALL died during he induction therapy. Infection was the most common cause of death.

CONCLUSIONThe outcome of patients treated with the 04 Protocol was favorable. Clinical risk classification and the leukemia cells of D19 are independent predictors of prognosis of ALL. High dose methotrexate played an important role in prevention and treatment of central nervous system leukemia. The mortality rate of this chemotherapeutic protocol during induction therapy was high.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; drug therapy ; Retrospective Studies ; Risk Factors ; Treatment Outcome

OBJECTIVETo establish the tridimensional culture method for tissue-engineered skin to observe the histomorphological change in human immortal KC strain (HacaT)cocultured with xenogenic acellular dermal matrix (ADM).

METHODSThe ADM was prepared from SD rats by a modified method. HaCaTs were cultured in defined KC-serum free medium. HaCaTs in log growth phase were inoculated on ADM at the cell density of 2 x 10(5)/cm(2). They were submergedly cultured for 5 days and then changed to air-liquid phase culture for another 5 days. ADM and growth of HaCaTs on day 1 and 5 after cocultured with ADM were observed with scanning electron microscope. The histological change in ADM and HaCaTs on day 1, 5, and 10 after cocultured with ADM were examined by HE staining.

RESULTSThe gross appearance of ADM was white with smooth and soft texture, and intact collagen bundles without cellular residue. HaCaTs adhered and stretched out pseudopodia on the surface of the ADM on day 1 after combined culture, and a monolayer of cells was formed on day 5, growing into 3-6 layers of cells on day 10 with a tendency to grow into ADM.

CONCLUSIONSSD rats ADM is benefit for the adhesion of HaCaTs and the permeation of nutrient solution, from which an engineered multiple-layered human skin can be obtained within 10 days.

Animals ; Cells, Cultured ; Coculture Techniques ; Dermis ; cytology ; Humans ; Keratinocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Skin, Artificial ; Tissue Engineering ; methods