Objective To investigate the effects of two inhibitors of arachidonic acid metabolic pathway (5-cyclooxygenase blockade MK886 and COX 2 blockade celecoxib) on growth and VEGF mRNA expression of human pancreatic cancer cell SW1990.MethodsPancreatic cancer cells SW1990 were cultured with different concentrations of MK886,celecoxib,MK886 and celecoxib,then the cell proliferation was detected by using CCK-8,BLT1 mRNA,PGE2 mRNA and VEGF mRNA expressions were determined by RTPCR.ResultsAfter 10 μmol/L MK886 or 20 mmol/L celecoxib treatment for 24 h,the growth of SW1990 was greatly suppressed ( 1.80 ±0.06 vs 1.65 ±0.10,2.04 ±0.03 vs 1.86 ±0.02,P ＜0.01 ),and the growth suppression of SW1990 cells was increased accompanying the raised concentration of MK886 or celecoxib.After both MK886 and celecoxib treatment for 12 h,the growth of SW 1990 cells was much obviously suppressed (1.72 ±0.05 vs 1.52 ±0.05,P ＜0.01 ).After celecoxib treatment for 48 h,the BLT1 mRNA,PGE2 mRNA and VEGFmRNA expressions were not significantly changed,but the expressions of PGE2 mRNA were significantly decreased ( P ＜ 0.05 ).After MK886 or MK886 + celecoxib treatment,the expressions of BLT1 mRNA,VEGF mRNA were significantly decreased ( P ＜ 0.05 ),but the expressions of PGE2 mRNA were not significantly changed when compared to control group.ConclusionsTwo metabolic pathways of arachidonic acid have a close relation with occurrence and proliferation of pancreatic cancer,when both of the pathways were blocked,the proliferation of the pancreatic cancer cell was suppressed obviously.

OBJECTIVE: To establish a method for whole genome amplification (WGA) based on (multiple displacement amplification MDA), and achieve DNA analysis of the human genome from low copy number (LCN). METHODS: DNA sample was used for WGA according to the REPLI-g kit protocol (QIAGEN, Germany). WGA product was used for DNA analysis according to the Applied Biosystems Profiler Plus kit protocol (ABI, USA). RESULTS: WGA product of DNA sample (10 pg) can be used for STR genotyping. CONCLUSION WGA technology could be helpful for LCN DNA analysis.
DNA/analysis* ; DNA Fingerprinting/methods* ; Genome, Human ; Genotype ; Humans ; Microsatellite Repeats ; Nucleic Acid Amplification Techniques/methods*

OBJECTIVEIn order to investigate whether the presence of distant metastases is associated with serum lipid abnormalities.

METHODSThe fasting serum lipid profile and various clinicopathological data of 324 breast cancer patients with and without synchronous distant metastases were collected and analyzed. The serum lipid profile, including total cholesterol (TC), triglycerides (TG), low-density (LDL-C) and high-density lipoprotein cholesterol (HDL-C) was determined. The nutritional status, the serum albumin was measured and body mass index (BMI) was calculated. Univariate analysis and multiple logistic regression analysis were carried out to investigate the association of serum lipid profile with distant metastases.

RESULTSUnivariate analysis showed that the distant metastasis rate was significantly higher in the breast cancer patients with an higher level of serum TC, TG, LDL-C, and LDL-C/HDL-C ratio (P < 0.05). Multiple logistic regression analysis showed that higher serum levels of TC, LDL-C and LDL-C/HDL-C ratio were independent risk factors for distant metastasis in breast cancer (OR = 2.324, 2.648 and 4.862, respectively).

CONCLUSIONSHyperlipidemia is significantly associated with the distant metastasis in breast cancer patients. Monitoring of serum lipid profile may be helpful to predict the occurrence of distant metastasis in breast cancer patients.

Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; Breast Neoplasms ; blood ; pathology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Humans ; Lipids ; blood ; Middle Aged ; Multivariate Analysis ; Neoplasm Metastasis ; Neoplasm Staging ; Nutritional Status ; Risk Factors ; Serum Albumin ; Triglycerides ; blood

OBJECTIVETo study the protection of Gan by using deoxyschizandrin (Wuzhi Capsule, WC) in the renal transplantation recipients while increasing the blood concentration of tacrolimus (Tac).

METHODSSeventy-three renal transplant recipients were randomly assigned to two groups, i.e., 35 in the WC group and 38 in the control group. All patients received Tac + MMF + Pred triple immunosuppressive therapy. WC was additionally given to patients in the WC group. One year was taken as one therapeutic course. Changes of the blood concentration of Tac were detected one week, 1, 3, 6, and 12 months after medication in the two groups using microparticle enzyme immune assay (MEIA). Meanwhile, the liver and kidney functions, and the blood glucose were tested.

RESULTSOne week after the application of WC, the blood Tac concentration increased 67.2% averagely. During the 1 -12 months of WC treatment, the Tac dosage was significantly lower in the WC group than in the control group (P<0.01). There was no significant difference in the liver and renal functions, or the blood glucose levels between the two groups (P>0.05).

CONCLUSIONWC could significantly increase the blood Tac concentration of renal transplant recipients, reduce their Tac dosages, with no obvious adverse reaction.

Adolescent ; Adult ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Graft Survival ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Postoperative Period ; Tacrolimus ; blood ; Treatment Outcome ; Young Adult

OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo investigate whether acute dipterex poisoning (ADP) may cause oxidative stress and free radical damage in the bodies of acute dipterex poisoning patients (ADPPs), and to explore the mechanisms by which ADP may cause oxidative stress and free radical damage.

METHODSFifty ADPPs and fifty healthy adult volunteers (HAVs) whose ages, gender and others were matched with the ADPPs were enrolled in a randomized controlled study, in which concentrations of nitric oxide (NO), vitamin C (VC), vitamin E (VE) and beta-carotene (beta-CAR) in plasma as well as concentration of lipoperoxide (LPO), and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and acetylcholinesterase (AChE) in erythrocytes were determined by spectrophotometric analytical methods.

RESULTSCompared with the average values of experimental parameters in the HAVs group, the average values of plasma NO and erythrocyte LPO in the ADPPs group were significantly increased (P<0.0001), while those of plasma VC, VE and beta-CAR as well as erythrocyte SOD, CAT, GPX and AChE in the ADPPs group were significantly decreased (P<0.0001). Bivariate correlation analysis and partial correlation analysis suggested that when NO and LPO values were increased, and VC, VE, beta-CAR, SOD, CAT and GPX values were decreased in the ADPPs, AChE value was decreased gradually in the ADPPs (P<0.001-0.0001). Reliability analysis of experimental parameters reflecting oxidative stress and free radical damage in the ADPPs showed that the reliability coefficient (8 items) alpha=0.6909, and the standardized item alpha=0.8574.

CONCLUSIONThe findings in the present study suggest that ADP can cause oxidative stress and free radical damage, and inhibit markedly erythrocyte acetylcholinesterase activity in ADPPs.

Acetylcholinesterase ; blood ; Adolescent ; Adult ; Ascorbic Acid ; blood ; Case-Control Studies ; Catalase ; blood ; China ; Cholinesterase Inhibitors ; poisoning ; Erythrocytes ; drug effects ; enzymology ; Female ; Free Radicals ; Glutathione Peroxidase ; blood ; Humans ; Insecticides ; poisoning ; Lipid Peroxides ; blood ; Male ; Nitric Oxide ; blood ; Oxidative Stress ; Poisoning ; blood ; Random Allocation ; Superoxide Dismutase ; blood ; Trichlorfon ; poisoning ; Vitamin E ; blood ; beta Carotene ; blood

OBJECTIVETo establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.

METHODSMTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured.

RESULTSJAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05.

CONCLUSIONJAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.

Annexin A5 ; analysis ; Apoptosis ; Cell Adhesion ; Cell Division ; Cell Line, Tumor ; Choriocarcinoma ; drug therapy ; genetics ; pathology ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology

OBJECTIVEThe protection rate of inoculation with BCG vaccine is only 50 percent, and most of patients with tuberculosis had a history of BCG vaccine inoculation. Adenosine (ADO) has an immunomodulating effect; it promotes immune reaction by increasing number of macrophage and enhancing phagocytosis. The present study was designed to investigate if combined use of adenosine with BCG enhances the anti-Mycobacterium tuberculosis effect of macrophage in mice.

METHODSFifty BALB/C mice were divided randomly into 3 groups: BCG group (n = 21), BCG plus ADO group (n = 21) and control group (n = 8). The mice in BCG and BCG plus ADO groups were inoculated with 0.1 ml BCG intradermally and the mice in BCG plus ADO group were injected intraperitoneally with ADO 30 mg/(kg.d) for 5 days. The mice in BCG group and control group were injected with NS 0.1 ml/d for 5 days. Six weeks after the last injection, all mice were challenged with intravenous 1 x 10(6) CFU human Mycobacterium tuberculosis virulent strain. After challenging, lung and spleen specimens were taken at the 10th, 20th and 30th days from the mice of BCG and BCG plus ADO groups and at the 30th day from mice in control group. The pathological examinations of lung and spleen sections were performed after HE staining and acid-fast staining, and detection of cell apoptosis was also performed.

RESULTSConsolidation with neutrophil infiltration was found in most of the lung tissue taken at the day 30; there were a lot of tuberculous granulomas and Mycobacterium tuberculosis in the lungs of control group. The alveolar septum in BCG gradually became wide and in interstitium lymphocyte infiltration dominated, and there were less tuberculous granulomas but there were large number of Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging. The widening of alveolar septum and consolidation of lung tissue in BCG plus ADO group became milder with monocytes infiltration, and there were few tuberculosis granulomas and Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging.

CONCLUSIONADO could increase the number of monocyte-macrophages and promoted anti-bacterial effects of these cells.

Adenosine ; administration & dosage ; immunology ; Animals ; BCG Vaccine ; administration & dosage ; immunology ; Disease Models, Animal ; Drug Therapy, Combination ; Injections, Intradermal ; Injections, Intraperitoneal ; Macrophages ; drug effects ; immunology ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Neutrophil Infiltration ; drug effects ; Phagocytosis ; drug effects ; Tuberculosis, Pulmonary ; immunology ; prevention & control

OBJECTIVESTo investigate CD3+CD56+ lymphocytes and their subsets in the peripheral blood of chronic hepatitis B patients and to explore the relationship between these cells and the pathogenesis of their diseases.

METHODSBlood samples from 53 chronic hepatitis B patients, 17 from HBV asymptomatic carriers (ASC) and 19 from healthy controls (HC) were collected. CD3+CD56+ lymphocytes were detected by flow cytometry (FCM), then the CD3+CD56+ lymphocytes were gathered to analyze their expressions of CD4, CD8, TCR Valpha24, TCRalpha/beta and TCRgamma/delta.

RESULTSThe number of CD3+CD56+ lymphocytes of chronic hepatitis B patients (7.4+/-4.6%) was more than those of ASC (4.5%+/-3.5%) and healthy controls (4.4%+/-3.7%). The expressions of TCR Valpha24 on CD3+CD56+ lymphocytes showed no significant differences among the three groups, but the expression of TCR Valpha24 on CD3-CD56+ lymphocytes of ASC ( 2.8%+/-1.4% ) was much more than that of the HC (1.7%+/-1.0%). For the subsets analysis, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of chronic hepatitis B (61.9%+/-16.8% and 68.1%+/-16.9%) were significantly higher than those of the HC (49.2%+/-15.6% and 56.4%+/-17.9%), while the TCRgamma/delta subsets of chronic hepatitis B and ASC (29.6%+/-15.4% and 30.5%+/-14.8%) were decreased significantly than those of the HC (41.4%+/-19.4%). On the other hand, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of severe chronic hepatitis B (69.0%+/-14.0% and 76.1%+/-12.9%) and CD8 subsets of moderate chronic hepatitis B patients (66.4%+/-14.9%) were significantly higher than those of the mild chronic hepatitis B patients (51.4%+/-16.2% and 62.1%+/-14.6%).

CONCLUSIONThe pathogenesis of chronic hepatitis B may positively relate to the high expression of CD8 on the CD3+CD56+ lymphocytes.

Adult ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

OBJECTIVETo assess the utility of P504S immunohistochemistry in the diagnosis and differential diagnosis of prostatic adenocarcinoma.

METHODSLight microscopy and immunohistochemistry examinations (EnVision staining) were performed in 117 cases of prostatic adenocarcinoma, PIN, AAH, ASAP, BPH and normal prostatic tissue to correlate the morphology and protein expression of P504S, 34betaE12, and P63.

RESULTSSeventy-one of the 78 (91%) cases of prostatic adenocarcinoma stained positive for P504S, with strong cytoplasmic granular staining in most cases, and a weak or intense granular staining along the circumferential luminal and apical cell border membrane in a few cases. Negative P504S immunostaining was observed in 7 of 78 (9%) cases of prostatic adenocarcinoma, all of which were clear cell type prostatic adenocarcinoma. Cases of PIN (9 cases), AAH (6 cases) and ASAP (2 cases) showed various expression levels of P504S. Sixty-five of 68 (96%) cases of normal prostates and BPH were negative for P504S and basal cell hyperplasia cases were also negative.

CONCLUSIONSP504S is a useful marker for microscopic diagnosis of prostatic adenocarcinoma, and immunohistochemistry study using a combination of P504S and 34betaE12/p63 may be of greater benefit in aiding the differential diagnoses.

Adenocarcinoma ; diagnosis ; DNA-Binding Proteins ; Diagnosis, Differential ; Genes, Tumor Suppressor ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Male ; Phosphoproteins ; analysis ; Precancerous Conditions ; diagnosis ; Prostate ; enzymology ; Prostatic Hyperplasia ; diagnosis ; Prostatic Intraepithelial Neoplasia ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Racemases and Epimerases ; analysis ; Trans-Activators ; analysis ; Transcription Factors ; Tumor Suppressor Proteins