Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.
Angelica ; chemistry ; Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cytarabine ; administration & dosage ; Humans ; K562 Cells ; Leukemia ; drug therapy ; Liver ; drug effects ; Male ; Mice ; Mice, SCID ; Polysaccharides ; administration & dosage

Objective To explore the distribution of water with high level arsenic and prevalence of arsenism along Huai'he River and the surrounding area of Hong'ze lake in Huai'an of Jiangsu. Methods Wate rsamples were collected and tested in 2008 from 18 villages of 6 towns according to history data in 3 counties like Xuyi,Jinhu and Hongze. Samples having arsenic level higher than 0.05 mg/L were investigated by epidemiological method and the patients were diagnosed by Standard of Diagnosis for Endemic Arsenism. Results All 5199 water samples were determined,and 260 water samples were exceeding the national drinking water quality level (0.05 mg/L) in 3 counties,the rates of exceeding diagnosis were 5.6%(247/4454),0.7%(4/597),6.0%(9/148) respectively. Total detected rate of endemic arsenic disease was 5.94%(128/2155). The detected rates of age group of 0 ～ ,20 ～,30 ～ ,40 ～ ,50 ～ ,60 ～ ,70 ～ ,80 ～ were 2.86%(1/35),2.11%(2/95),1.26%(3/239),3.10%(16/516),5.53% (32/579),10.07%(41/407),11.84%(27/228),10.71%(6/56) respectively. The detected rate of male (9.10%,78/857) was higher than that of female(3.85%,50/1298,χ~2 = 25.46,P < 0.01). Conclusions Huai'he River and the surrounding areas of Hong'ze lake like Xuyi,Jinhu and Hongze are identified existing endemic arsenic disease area. The prevention of arsenism should be strengthened in these areas.

To explore the protective effect of Angelica sinensis polysaccharides(ASP) on subacute renal damages induced by D-galactose in mice and its mechanism. Male C57BL/6J mice were randomly divided into 3 groups, with 10 mice in each group. The D-galactose model group was subcutaneously injected with D-galactose (120 mg x kg(-1)), qd x 42; the ASP + D-galactose model group was intraperitoneally injected with ASP since the 8th day of the replication of the D-galactose model, qd x 35; and the normal control group was subcutaneously injected with saline at the same dose and time. On the 2nd day of after the injection, the peripheral blood was collected to measure the content of BUN, Crea, UA, Cys-C; paraffin sections were made to observe the renal histomorphology by HE staining; senescence-associated β-g-alactosidase (SA-β-Gal) stain was used to observe the relative optical density (ROD) in renal tissues; transmission electron microscopy was assayed to observe the renal ultrastructure; the renal tissue homogenate was prepared to measure the content of SOD, GSH-PX, MDA; the content of AGEs and 8-OH-dG were measured by ELISA. According to the result, compared with the D-galactose model group, the ASP + D-galactose model group showed obviously decreases in the content of BUN, Crea, UA, Cysc, AGES, 8-OH-dG, the number of hardening renal corpuscle, renal capsular space and renal tubular lumen, ROD of SA-β-Gal staining positive kidney cells, mesangial cells, basement membrane thickness, podocyte secondary processes fusion and MDA and increases in the number of normal renal corpuscle, ribosome and rough endoplasmic reticulum in podocytes, the activity of SOD and GSH-PX. In Conclusion, A. sinensis polysaccharides can antagonize kidney subacute damages induced by D-galactose in mice. Its protective mechanism may be correlated with the inhibition of the oxidative stress injury.
Angelica sinensis ; chemistry ; Animals ; Deoxyguanosine ; analogs & derivatives ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Galactose ; adverse effects ; Humans ; Kidney ; anatomy & histology ; drug effects ; injuries ; Kidney Diseases ; chemically induced ; drug therapy ; metabolism ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; Oxidative Stress ; drug effects ; Polysaccharides ; administration & dosage ; Protective Agents ; administration & dosage

OBJECTIVETo explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.

METHODAcute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.

RESULTThe purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.

CONCLUSIONASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.

Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Neoplastic Stem Cells ; cytology ; drug effects ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects

BACKGROUNDAbdominal pain is a common symptom among patients with acute appendicitis, yet these patients have long been denied relief from suffering because of widespread misconceptions associated with the use of opioids. We determined whether morphine hydrochloride masked the physical signs in adults with acute appendicitis and assessed the efficacy of morphine in relieving abdominal pain.

METHODSA prospective, double-blind, placebo controlled, clinical trial was conducted with 106 adult patients between 16 and 70 years old with acute appendicitis. Patients were randomly divided into a morphine group (n=54) or a normal saline group (n=52). All patients presented with acute abdominal pain with onset within 3 days. The morphine group received hypodermic injection of morphine (0.15 mg/kg; maximum 20 mg) and the control group members were given an equivalent volume of normal saline solution. The clinical symptoms, physical signs, and patients' cooperation during physical examination were assessed before and after 30 minutes of morphine or normal saline administration.

RESULTSAbdominal pain was significantly relieved and the patients' cooperation was improved in the morphine group after 30 minutes treatment compared with the control group and before morphine administration (P<0.05). The physical signs were unaffected by either treatment (P>0.05).

CONCLUSIONSMorphine relieved abdominal pain and improved the patients' cooperation for treatment and care. Furthermore, the morphine did not mask the physical signs of acute appendicitis.

Abdominal Pain ; drug therapy ; pathology ; Adolescent ; Adult ; Aged ; Analgesics, Opioid ; therapeutic use ; Appendicitis ; drug therapy ; pathology ; Female ; Humans ; Male ; Middle Aged ; Morphine ; therapeutic use ; Young Adult

Objective To improve the CR image quality and to promote the digital image standard constitution by analyzing the common problems and malpractiees in application of computed radiography. Methods Phenomenon and reasons of 107 CR junk-films from nine three-"A"-hospitals were analyzed, discussed,recorded,and statistised by 20 radiologists,radiographers and engineers.Results Among 107 junk fihns,there are 36 cases(33.64%)of incorrect operations,29 cases(27.10%)of artifacts in reading and transferring the data of IP,15 cases(14.02%)of artifacts in IP system,and 13 cases (12.15%)of selection of inappropriate radiographic parameters,and 9 eases(8.41%)of printer-failures, and 5 eases(4.67%)of inappropriate post-processing techniques.By analyzing the reasons of 107 junk films we found that 60.74% were due to less responsibilities and incorrect operations,and 35.51% were due to new problems in CR techniques,and other were due to inappropriate post-processing techniques. Conclusion Responsibilities,operation regulations,digital image quality standards,studying of new techniques and appropriate use of the post-processing techniques are the key points for improving the CR image quality and the diagnosis level.

Thrombotic microangiopathy (TMA) is a lethal transplantation-associated complication which exactly likes acute intestinal graft-versus-host disease (GVHD) in the clinical manifestation. 373 consecutive patients with hematological diseases received family HLA matched or mismatched HCT from May, 2002 to July, 2004. To analyse the clinical and pathological characteristics of TMA, 30 patients who suffered from severe diarrhea and received colonoscopic examination and gut biopsy were retrospectively analyzed. The results indicated that 7 patients originally diagnosed as gut GVHD showed the pathological evidence of enteric TMA. The incidence of TMA was 7 out of 30 specimen (23.3%). Pathological evidence of enteric TMA shown microvascular disorder characterized by thrombus in the capillary without infiltration of lymphocytes and perivascular hemorrhages in the mucosa, swelling and focal denudation of epithelial cells. All patients with TMA were associated with cytomegalovirus (CMV) antigenemia/disease. Among these patients, 4 cases, who only showed TMA without the evidence of gut GVHD pathologically, displayed treatment-resistant bloody diarrhea, renal failure, veno-occlusive disease, hemorrhagic cystitis, hemolytic anemia as well as thrombocytopenia. But the other 3 cases, with co-existence of both TMA and GVHD pathological characteristics had better treatment response. Survival analysis indicated that 3 patients with TMA-GVHD survived for 461 to 536 days but three out of four TMA patients died from VOD with liver failure as well as multiple organ failure during 101 to 254 days after HCT. In conclusion, to better diagnose those patients with severe and refractory diarrhea following HCT, pathological examination may indicate crux evidence to identify intestinal TMA from gut GVHD. Furthermore, this primary report has first evidenced that TMA and TMA-GVHD are two pathologically well-recognized subtypes with the difference between the pathological characteristics, treatment response and clinical outcomes.
Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Intestinal Diseases ; diagnosis ; etiology ; pathology ; Purpura, Thrombotic Thrombocytopenic ; diagnosis ; etiology ; pathology ; Reference Standards ; Retrospective Studies ; Thrombosis ; diagnosis ; etiology ; pathology

OBJECTIVETo summarize the clinical characteristics, diagnosis and surgical in-treatment results of congenital coronary artery fistulas (CAF) in adults.

METHODSFourteen patients (8 men, 6 women), aged from 18 to 60 years with a mean of 32 +/- 13 years, underwent surgical correction of CAF between March 1985 and April 2002. Eleven of the 14 patients (78.57%) were symptomatic. The diagnosis of CAF was made by echocardiography or angiocardiography preoperatively. The fistulae originated from the right, left and double coronary arteries in 10 (71%), 3 (21%) and 1 (7%) patient(s), respectively. The fistulae drained into the right ventricle (8 patients), left ventricle (4), right atrium (1) and pulmonary artery (1), respectively. The diameter of fistulae ranged from 0.30 to 1.80 cm with a mean of (1.16 +/- 0.49) cm. There were 6 CAF patients associated with coronary artery aneurysms and 4 CAF patients with other coexisting cardiac defects. The distal fistulae were closed in 10 patients with cardiopulmonary bypass (CPB) and 4 patients without CPB. The coexisting defects were corrected simultaneously.

RESULTSThere was no early and late death. One patient had low cardiac output syndrome and cured during early postoperative period. Twelve patients (85.71%) were followed up for a mean period of 3.35 +/- 4.28 years without myocardial ischemia or infarction and recurrent fistulae. Heart function was improved to NYHA functional class I in 11 patients and class II in 1 patient.

CONCLUSIONSAll adult patients with CAF who have demonstrable hemodynamic and cardiovascular morphological changes should be surgically treated as early as possible. The appropriate surgical management and reliable myocardial protection are key points of good surgical results.

Adolescent ; Adult ; Coronary Vessel Anomalies ; diagnosis ; surgery ; Female ; Fistula ; congenital ; diagnosis ; surgery ; Humans ; Male ; Middle Aged

BACKGROUNDThe multidrug resistance (MDR) associated with the expression of the mdr1 gene and its product P-glycoprotein is a major factor in the prognosis of hepatocellular carcinoma cell (HCC) patients treated with chemotherapy. Our study was to establish a stable HCC MDR cell line where a de novo acquisition of multidrug resistance specifically related to overexpression of a transgenic mdr1.

METHODSThe 4.5-kb mdr1 cDNA obtained from the plasmid pHaMDR1-1 was cloned into the PCI-neo mammalian expression vector, later was transferred by liposome to human hepatocarcinoma cell line HepG2. Then the transfected HepG2 cells resisting G418 were clustered and cultured and the specific fragment of mdr1 cDNA, mRNA and the P-glycoprotein (Pgp) in these HepG2 cells were detected by PCR, RT-PCR and flow cytometry, respectively. The accumulation of the daunorubicin was determinated by flow cytometry simultaneously. The nude mice model of grafting tumour was established by injecting subcutaneously HepG2/mdr1 cells in the right axilla. When the tumour diameter reached 5 mm, adriamycin was injected into peritoneal cavity. The size and growth inhibition of tumour were evaluated.

RESULTSThe mdr1 expression vector was constructed successfully and the MDR HCC line HepG2/mdr1 developed. The PCR analysis showed that the specific fragment of mdr1 cDNA in HepG2/mdr1 cells, but not in the control group HepG2 cells. Furthermore, the content of the specific fragment of mdr1 mRNA and Pgp expression in HepG2/mdr1 cells were (59.7 +/- 7.9)% and (12.28 +/- 2.09)%, respectively, compared with (16.9 +/- 3.2)% and (3.07 +/- 1.06)% in HepG2 cells. In the nude mice HCC model, the tumour genes of both groups were identified. After ADM therapy, the mean size of HepG2 cell tumours was significantly smaller than HepG2/mdr1 cell tumours.

CONCLUSIONThe approach using the transfer of mdr1 cDNA may be applicable to the development of MDR hepatocarcinoma cell line, whose MDR mechanism is known. This would provide the experimental basis of MDR research.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Animals ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; therapeutic use ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Flow Cytometry ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; genetics ; pathology ; Mice ; Mice, Nude ; Mitomycin ; pharmacology ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction ; Xenograft Model Antitumor Assays ; methods

Objective To evaluate the irnmunogenicity, safety and stability of the manufacture process regarding three consecutive lots of influenza split vaccines (Anflu ). Methods A double-blind, randomized and controlled clinical trial was conducted in healthy volunteers. A total of 566 subjects aged 18 to 60 years were recruited and stratified into four age groups before randomly assigned into four groups. Each group would receive one dose of influenza vaccine from either one of the three lots ofAnflu or one lot of the licensed control vaccine. Each dose of the vaccines contained 15 μg of each of the H1N1, H3N2 and B type antigen. Safety was assessed through 30-minute observation for immediate allergic reaction and three-day observation after vaccination. HI antibody titers were determined before vaccination and on day 21, after vaccination. Results Mild adverse reaction was reported and the overall incidence rates on fever of the four groups were from 1.4% to 2.8% but no significant difference was observed between groups. Seroconversion rates of the three viral strains in four groups were 80.3% and above with fold increase as≥11.1 and protection rate was≥93.4%. For the three lots of investigated vaccines, all of the indexes of the three viral strains in four groups exceeded the standards on EMEA and FDA for influenza vaccine. Conclusion The three consecutive lots of Anflu appeared to be good, with both consistent immunogenieity and safety, indicating the stability of manufacture process.