Triterpenoid saponins from Stauntonia chinensis ameliorate insulin resistance via the AMP-activated protein kinase and IR/IRS-1/PI3K/Akt pathways in insulin-resistant HepG2 cells.

Inflammation and oxidative stress play crucial roles in the etiology of type 2 diabetes mellitus. In this study, we examined the anti-diabetic effects of triterpenoid saponins extracted from Stauntonia chinensis on stimulating glucose uptake by insulin-resistant human HepG2 cells. The results showed that saponin 6 significantly increased glucose uptake and glucose catabolism. Saponin 6 also enhanced the phosphorylation of AMP-activated protein kinase (AMPK) and activated the insulin receptor (IR)/insulin receptor substrate-1 (IRS-1)/phosphoinositide 3-kinase (PI3K)/Akt pathway. Therefore, our results suggest that saponins from S. chinensis improve glucose uptake and catabolism in hepatic cells by stimulating the AMPK and the IR/IRS-1/PI3K/Akt signaling pathways. The results also imply that saponins from S. chinensis can enhance glucose uptake and insulin sensitivity, representing a promising treatment for type 2 diabetes mellitus.

[The assay of intracelluar cytidine deaminase activity and its clinical significance in patient with acute leukemia].

OBJECTIVE: To investigate the relationship between the intracellular cytidine deaminase (CDD) activity and drug resistance in acute leukemia (AL) of different stage. METHODS: CDD activity of 80 AL patients was determined by radioactive isotope assay. RESULTS: CDD activity of previously untreated and refractory/relapse AL patients was much higher than that of patients in complete remission (P<0.05). There was no significant difference of CDD activity between previously untreated and refractory/relapse patients (P>0.05). The activity of CDD had no correlation with age, sex, classification of AL and effect of chemotherapy containing cytidine deaminase (Ara-C). CONCLUSION: Intracellular CDD activity in AL patients may be related to the stages of the disease. It seemed that CDD might not be considered as an independent predictive factor for the drug healing efficacy of Ara-C.

[Diffuse alveolar hemorrhage as a complication in patients with hematologic malignancies after chemotherapy: report of two cases and literature review].

OBJECTIVE: To study diffuse alveolar hemorrhage (DAH) in patients with hematologic malignancies after chemotherapy and discuss the possible etiology and appropriate therapy. METHODS: Symptoms, physical examinations, laboratory examination, chest radiographs or computed tomographic (CT) scans, treatments and outcomes of two patients with acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL) each after chemotherapy were presented. RESULTS: Both of the patients developed cough, progressive dyspnea, a drop of hemoglobin level, hypoxemia and widespread pulmonary infiltrate on chest radiographs or CT scans after chemotherapy. Moreover, case 1 (ALL) had high fever and bloody fluid drained from the intubation of mechanical ventilation, case 2 (NHL) developed continual hemoptysis. They were diagnosed as DAH and improved significantly after intermediate- or high-dose corticosteroid therapy. CONCLUSIONS: DAH is a rare fatal acute noninfectious pulmonary complication in patients with hematologic malignancies after chemotherapy. Early accurate diagnosis, identifying the underlying cause and appropriate treatment are critical for the management of DAH.

Measurement of electrolyte concentrations in expressed prostatic secretion and urine from patients with chronic prostatitis and its implications.

This study was aimed at measuring concentration of electrolytes, especially K+ in expressed prostatic secretion (EPS) and urine from patients with chronic prostatitis. The concentration of potassium, sodium, chloride, calcium in EPS and urine of 31 controls and 79 patients with prostatitis were measured and analyzed. There was no significant difference in the concentrations of potassium, sodium, chloride and calcium between the patients and the controls. Among the patients treated effectively, potassium concentration was 40.66 +/- 17.10 mmol/l before treatment and 33.42 +/- 17.27 mmol/l after treatment. While among the patients treated ineffectively, potassium concentration was measured as 37.57 +/- 16.93 mmol/l and 50.66 +/- 18.77 mmol/l before and after treatment respectively. The concentrations of electrolytes in prostatic fluid varied greatly between individuals. Potassium concentration in EPS decreased significantly after treatment among the patients with obvious treatment effectiveness, while increased among those who failed the treatment. EPS potassium concentration was also found to be lower in patients with pain than those without pain. No significant difference was found between the normal group and the no-pain patients.

[Studies of treatment strategy and prognosis on acute myeloid leukemia with chromosome 8 and 21 translocation].

OBJECTIVE: To investigate the relationship between the biological features and the treatment efficacy and prognosis in acute myeloid leukemia subtype M2 (AML-M2) patients with chromosome 8 and 21 translocation. METHODS: By using Cox regression model and Kaplan-Meier analyses, prognostic factors in 54 cases of de novo adult AML with t(8;21) in our institute from 1990 to 2003 were retrospectively analyzed. RESULT: The complete remission (CR) rates were 81.9% for all M2 patients, 82.4% for patients with normal karyotype, 88.5% for patients with t(8;21) [P > 0.05 for normal karyotype vs t(8;21)], 100.0% for 28 patients with t(8;21) alone and 75.0% for 24 patients with additional chromosome abnormalities (P < 0.01). The actuarial 3 year overall survival(OS) was 26% for M2 patients with normal karyotype, 25% for patients with t(8;21) [P > 0.05 for normal karyotype vs t(8;21)], in whole t(8;21) group, 46.4% for patients with t(8;21) alone and 0% for patients with additional chromosome abnormalities (P < 0.01). Multivariate analysis of prognostic factors showed that chromosome abnormalities besides t(8;21) was the only factor affecting CR, disease-free survival (DFS) and OS. DFS of allogeneic hematopoietic stem cell transplantation (HSCT) and intermediate-dose cytarabine/high dose cytarabine (IDAC) groups were better than the group received routine dose cytarabine as postremission therapy (P < 0.01). CONCLUSION: AML with t(8;21) is not a single defined AML subset, and patients with additional chromosome abnormalities have a worse prognosis. HSCT and IDAC could improve the outcome. HSCT is the best choice for patients with high risks, especially with additional chromosome abnormalities.

[Treatment of refractory and relapsed acute lymphocytic leukemia in adults].

OBJECTIVE: To analyze on the efficacy and toxicity of fludarabine and teniposide + mitoxantrone (MIT) regimens on treating refractory and relapsed acute lymphocytic leukemia in adult patients. METHODS: Teniposide 100 mg/d, 5-7 d, MIT 10 mg/d, 2 d and fludarabine regimens [Flu 30 mg/(m(2) . d), 3- 5 d, Cytarabine (Ara-c )1-2 g/(m(2) . d), 5 d; Flu 50 mg/d, 5 d, Ara-c 200 mg/d, 5 d, MIT 4 mg/d, 4 d] were used to treat 42 cases of adults with refractory and relapsed acute lymphocytic leukemia(ALL). G-CSF 5 microg/(kg . d) were used when WBC<1.0 x 10(9)/L. RESULTS: In both the regimens fludarabine and VM (teniposide + MIT), the complete remission (CR) rate was 45% versus 31.8% (P>0.05); the median neutropenia began 6 days after the regimens arresting and lasting 10 versus 7.5 days, P>0.05; thrombocytopenia begin at time of 10 versus 6.5 days (P<0.05) after the regimens arresting and lasting 6 versus 10 days (P>0.05). Fludarabine regimen had less non-haematological toxic effect than that of VM. CONCLUSION: Compared with VM, Fludarabine regimen was a very effective alternative treatment for CR induction in adult patients with refractory and relapsed ALL and low toxicity.

[Clinical study of Philadelphia chromosome-positive acute lymphoblastic leukemia].

OBJECTIVE: To study the clinical characteristics and therapeutic outcome of Ph+ acute lymphoblastic leukemia (ALL). METHODS: Thirty previously untreated cases of Ph+ B-ALL were diagnosed in our institute. The patients were treated with combination chemotherapy of CODP +/- L regimen, Imatinib (400 approximately 600 mg/d) was continuously given to those who couldn't reach CR. Fourteen patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CR, while 16 received consolidation of intensive chemotherapy. RESULTS: Thirty (32.6%) of 92 ALL patients were diagnosed as Ph+ ALL, with a median age of 25.5 (14 - 60). Among them Ph+ as the sole anomaly was seen in 16 patients, and Ph+ with additional chromosome abnormalities in 14. Besides the B cell markers, 23 (76.7%) patients had CD34+ and 13 (43.3%) CD13+ and/or CD33+. Nineteen of the Ph+ ALL patients underwent molecular analysis; 13 (68.4%) expressed P190 and 6 (31.6%) P210. Increased WBC (> 30 x 10(9)/L) was found in 22/30 cases while WBC > 100 x 10(9)/L in 9/30 cases. The chemotherapy complete remission rate was 68.8% in patients with only Ph+ versus 28.6% in those with additional chromosome abnormalities. All seven refractory/relapsed patients reached CR with Imatinib therapy. The total complete remission rate was 73.3% in all Ph+ ALL patients. The median remission duration was shorter in patients with additional chromosome than in those with only Ph+ (1 vs 7 months, P < 0.05), and so was the survival period (7 vs 9 months, P > 0.05). The remission duration was significantly longer in patients received allo-HSCT than in those received chemotherapy only (8 vs 0.5 month, P < 0.05), and so was the survival period (12.5 vs 6 months, P < 0.05). CONCLUSION: Additional chromosome abnormalities negatively affect the prognosis and therapeutic effect of Ph+ ALL patients. Imatinib is effective for the induction therapy of Ph+ ALL. The survival period of patients who received allo-HSCT was obviously longer than those who received chemotherapy only.

[Cytogenetic and clinical study of Philadelphia chromosome positive adult acute leukemia].

To explore the cytogenetics and related clinical characteristics of adult acute leukemia with Philadelphia chromosome positive (Ph(+)AL), MIC classification by morphology, immunology and cytogenetics was used to retrospectively study 79 patients with Ph(+)AL hospitalized in the Institute of Hematology, People Hospital in Beijing from October 1991 to September 2003. The results showed that 6.9% cases were diagnosed as Ph(+)AL and classified into three subtypes: acute lymphoblastic leukemia (Ph(+)ALL) in 56 patients (18%), acute myeloid leukemia (Ph(+)AML) in 10 patients (1.2%) and mixed acute leukemia (Ph(+)MAL) in 13 patients. B-cell antigen expression was found in 52 out of 56 patients with Ph(+)ALL. 54.4% (43/79) patients had additional chromosome abnormalities including chromosome 7, double Ph and plus 8, etc. Complete remission (CR) rate of Ph(+)ALL and Ph(+)MAL was 57.0%, none of Ph(+)AML achieved CR. Median overall survival of Ph(+)ALL, Ph(+)MAL and Ph(+)AML were 10, 10 and 2.5 months respectively. It is concluded that Ph(+)AL has highly heterogeneity involving various differentiated stages of immature leukemic cells. Since the poor prognosis associated with this kind of AL, early diagnosis with MIC classification is a prerequisite to take more effective conditioning regimen and prospectively consideration of allogeneic stem cell transplantation to improve prognosis.

[MICM classification and prognostic analysis of 80 adolescents with acute lymphoblastic leukemia].

OBJECTIVES: To explore MICM classification and adverse prognostic factors in adolescents with acute lymphoblastic leukemia (ALL). METHODS: The MICM classification, clinical characteristics of 80 adolescents with ALL admitted to our hospital from January 1998 to December 2002 were retrospectively analyzed. Survival data were estimated by the Kaplan-Meier method and the prognostic factors were analyzed with the COX regression model. RESULTS: In the 80 patients, B-ALL and T-ALL accounted for 69.12% and 26.47%, respectively. The percentage of Ph(+)ALL was 18.37% (9/49), and that of hyperdiploidy was 4.08%. Patients at diagnosis with high leukocyte counts (> 50 x 10(9)/L) accounted for 27.94%. Among the 78 cases treated with VDP(L) or CODP(L) regimens, 73 (91.03%) obtained CR in 4 weeks. After a median follow-up of 24 months, the estimated 3-year disease-free survival (DFS) rates of patients receiving chemotherapy or allo-HSCT were (32.55 +/- 16.50)% and (69.58 +/- 8.72)%, respectively (P < 0.05). In COX analysis, high initial leukocyte counts (> 50 x 10(9)/L) and Philadelphia chromosome positivity were adverse prognostic factors for long-term survival. CONCLUSIONS: MICM classification has important clinical and prognostic significance in the risk-directed therapy of adolescents with ALL. The adverse prognostic features for these patients were high leukocyte counts, less incidence of chromosome hyperdiploidy and Ph chromosome positivity.

An epidemiological study of patients with chronic prostatitis.

OBJECTIVE: To assess possible causes, clinical symptoms and improvements in treatment for chronic prostatitis in China. PATIENTS AND METHODS: The study comprised 3000 patients with chronic prostatitis (aged 20-59 years), selected from urological clinics at province, city and county levels in Anhui (a province in mid-eastern China). Anonymous questionnaires were distributed which included 29 items to ascertain patient age, height, weight, educational background, personality, career, disease course, treatment status, prostatic fluid test and score of the National Institute of Health Chronic Prostatitis Symptom Index (NIH-CPSI). RESULTS: In all, 2498 valid questionnaires were collected (response rate 83.3%); 78.2% of the patients were aged <40 years and there were fewer patients in groups of increasing age. Discomfort and pain in the pelvis was reported by 52.3%, pain on urination by 23.0%, sexual discomfort by 21.8%, urinary frequency by 65.8%, and voiding discomfort by 74.4%; 34.9% of men were satisfied with their previous treatment. CONCLUSION: In China there are fewer patients with chronic prostatitis as age increases. The main symptoms were voiding dysfunction, e.g. frequent urination. The prevalence of pain, e.g. on urination, was significantly less than documented in other parts of the world. Most patients had used antibiotics to treat their prostatitis; generally the effect of treating prostatitis was unsatisfactory.

[Expression of cytidine deaminase mRNA in bone marrow cells from patients with acute leukemia].

The aim of the study was to determine the expression of cytidine deaminase (CDD) gene in bone marrow cells from patients with acute leukemia (AL) and evaluate the relationship between CDD expression and clinical feature. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used for detection of expression level of CDD mRNA in bone marrow cells from 83 patients with acute leukemia and from 15 healthy peoples as control. CDD/beta-actin ratio >or= 0.5 was considered to be positive. The results showed that expression levels of CDD of 31 previously untreated patients were higher than those of 23 cases of AL in complete remission and of normal controls. Expression levels of CDD of 29 relapse/refractory patients were also higher than those of 23 AL patients in complete remission and of normal subjects. The expression levels of CDD in relapse/refractory ALL were higher than those in AML while expression levels of CDD were not correlated with the outcome of therapy. It is concluded that the level of CDD mRNA expression varies at the different stage of acute leukemia. The expression level of CDD seems not to be a prognostic factor.

[The identical effects of B7-1 and B7-2 on regulation of human IL-2 gene transcription factors NF-kappa B and AP-1].

To detect effects of B7 co-stimulation on cytokines, especially on IL-2 mRNA and transcription factors NF-kappa B and AP-1, antiB7-1 McAb, antiB7-2 McAb and C TLA-4 Ig were added into mixture lymphocyte reaction (MLR) system to block B7/C D28 signal transduction, IL-2 mRNA and IL-4 mRNA were determined by using competitive PCR and IFN-gamma mRNA by using semi-quantitative PCR. MHC class II molecules and B7 transfectants were used to stimulate CD28(+) T cell, effects of B7 on NF-kappa B and AP-1 were detected by DNA-protein binding assay. The results showed that IL-2, IL-4 and IFN-gamma mRNA were significantly lower when blockade of B7-2 in MLR than blockade of B7-1. Synergistic effects could be seen with combination of two or three antibodies. One to six hours after MLR, tDR7 alone induced NF-kappa B binding activity; cotransfecting B7 no significantly difference at early time point. After 6 hours, induction of tDR7 was decreased whereas B7 prolonged the induction of NF-kappa B till 72 hours. tDR7 alone also upregulated AP-1 binding activity, on the contrary to NF-kappa B, co-transfecting B7-1 and B7-2 decreased AP-1 binding activity within 24 hours. But during 48 - 72 hours, B7 also prolonged the AP-1 binding activity. It is concluded that after MLR, B 7 increased IL-2 secretion by decreasing the degradation of IL-2 mRNA and upregulating IL-2 transcription factors. B7 also induced several kinds of cytokines secretion. Effects of B7-1 and B7-2 had no significant difference on transcription factors. It is suggested that the different functions between B7-1 and B7-2 maybe related to the difference of cell expression and kinetics. To study the molecular mechanism of B7 mediated T cell immune tolerance can help us to design a better clinic schema to prevent transplantation rejection and GVHD.

[Transfer and Expression of Recombinant Human Thrombopoietin Gene in COS-7 Cells and Mice In Vivo]

In order to investigate the expression of recombinant TPO gene in COS-7 cells and in vivo of the mouse model, eukaryotic expressing plasmid pcd2/TPO with human TPO cDNA was constructed with DNA recombinant techniques. The plasmid pcd2/TPO was transiently transfected into the COS-7 cells by means of lipofection, the naked pcd2/TPO plasmid was injected into the skeletal muscle of mice with electric pulses. RT-PCR and ELISA methods were used to detect the TPO expression of the transfected COS-7 cells, both showed high level expression. The MTT test showed the expressed TPO had proliferative activity to TPO-dependent cell line. High efficiency of gene transfer in transgenic mice was also observed by RT-PCR and immunohistochemical methods. The serum TPO level [(1 185 +/- 264) ng/L] in transgenic mice was quite different compared with the normal mice [(250 +/- 76) ng/L]. All these results provided solid foundations for the research of TPO gene therapy in the future.