Objective: To investigate the promoter region CpG island methylation status of glutathione S-transferase p1 (GSTP1) gene in prostate carcinoma (PCa) tissues and to search for new molecular markers for the early diagnosis of PCa. Methods: The promoter region CpG island methylation status of 31 PCa, 18 benign prostatic hyperplasia tissues (BPH) and 3 normal prostate tissues (NP) were examined by nest methylation-specific PCR (NMSP) technique, cloning and sequencing. Results: NMSP showed that the methylation rates of PCa, BPH and NP tissues were 83%, 0% and 0%, respectively. The CG sites methylation rates of PCa, BPH and NP tissues were 96%, 34% and 37%, respectively as determined by cloning and sequencing (P<0.01), with no significant difference found between those of BPH and NP tissues(P>0.05). A combination of NMSP and prostate-specific antigen (PSA) was better than PSA alone in screen of PCa. Conclusion: CpG islands of promoter region of GSTP1 gene is hypermethylated in PCa tissues. NMSP is sensitive and specific in detection of GSTP1 methylation, which may serve as a new method for early screen of prostate carcinoma.

AIM To determine the dynamic changes of monoamine neurotransmitters and their metabolites in various brain regions of cerebral ischemia reperfusion mice. METHODS Concentrations of monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT) and metabolites were determined by HPLC-ECD on d 0,1,3,5 and d 20 after cerebral ischemia reperfusion by common carotid artery occlusion. RESULTS The cerebral ischemia reperfusion mice showed decreased concentrations of NE, MHPG, DA, DOPAC, 5-HT and 5-HIAA in various brain regions, especially in hippocampus. CONCLUSION Several neuron systems play an important role in neurons damage of cerebral ischemia reperfusion, especially the NE and DA in hippocampus which is sensitive to the ischemia damage. The data offer useful guides for clinical treatments of cerebral ischemia diseases.

BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot.
OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s.
METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method.
RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo evaluate the clinical efficacy and side effects of Shelian capsule (SLC) as adjuvant of embolismic chemotherapy on primary hepatic carcinoma.

METHODSOne hundred and twenty patients with hepatic carcinoma were conducted arterial embolismic chemotherapy with FAM program for two therapeutic cycles. The 60 patients in the treated group were taken SLC orally for auxiliary treatment, and the other 60 in the control group were given glucurolactone instead. The tumor size, Karnofsky score, clinical symptoms, alpha fetoprotein (AFP), NK cell, T cell-subgroup and adverse effect before and after treatment between the two groups were compared respectively.

RESULTSAs compared with the control group, in the treated group the Karnofsky score was better ((P < 0.05) and the remission rate of clinical symptom were superior (66.7% vs 88.3 %, P < 0.05). The ratio of CD4/CD8 and NK cell activity was significantly different between the two groups (P<0.05). There was no obvious adverse effect in the treated group.

CONCLUSIONSLC can effectively enhance the immunity of patients with tumor, remit clinical symptoms, improve quality of life and without any side effects.

Adjuvants, Immunologic ; therapeutic use ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; Capsules ; Carcinoma, Hepatocellular ; immunology ; therapy ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Doxorubicin ; administration & dosage ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fluorouracil ; administration & dosage ; Humans ; Liver Neoplasms ; immunology ; therapy ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Phytotherapy

Animals ; Aorta ; pathology ; Apolipoproteins E ; deficiency ; Atherosclerosis ; metabolism ; pathology ; Female ; Glycosides ; isolation & purification ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To study the outcomes of selective reduction of triplet pregnancy by assisted reproductive technology. Methods The clinical data of 31 women who succeeded in conception by vitro fertilization-embryo transfer in the third affiliated hospital of Guangzhou Medical University were retrospectively investigated to analyze and compare the rates of abortion from triplet pregnancy, twin pregnancy and single pregnancy after selective reduction of triplet pregnancy, the incidence of pregnant complications, outcomes of perinatal period. Results There were no significant differences between triplet pregnancy and the twin and single pregnancy after selective reduction of triplet pregnancy in terms of pregnancy and parity time, fetal disease, premature rupture of membrane, severe eclampsism, gestational diabetes and postpartum hemorrhage (P>0.05). There were significant differences in gestational weeks, birth weight, rate of premature birth, rate of neonatal transfer to NICU and neonatal RDS: The gestational time in the triplet group longer than the groups of twin pregnancy and single pregnancy after selective reduction of triplet pregnancy (P<0.05). (37.3 ± 1.9) vs. (35.2 ± 0.9), (32.6 ± 2.3), respectively), the rate of premature birth dropped (100%vs. 100%, 33.3%, respectively), the body weight was increased (1 707 ± 360.4)g vs. (2 066.1 ± 307.5)g, (2 712.5 ± 514.1)g, respectively and the neonatal complication rate was reduced (P<0.05). Conclusion The selective reduction of multiple pregnancy may decrease the risk of premature birth and reduce the rate of lower body weight of neonates, improving the pregnancy outcomes, but the higher rate of abortion at the early or middle course of pregnancy after selective reduction is worth our attention.

Objective To report a 33-year-old man with post-headache stroke-like episodes,with whom ischemic changes were found in basal ganglia and occipital-temporal lobe and muscle biopsy revealed abnormal mitochondrial structure and function without regular mutations detected in mtDNA.Methods Gene chip technique was used to detect the mutation of whole sequence of mtDNA,and direct sequencing technique was used to confirm the mutations.Results Three mutations were found.A new mutation in the mitochondrial cytochrome C oxidase subunit 1 (MTCO1),a T→C transition at nucleotide position 6253 resulted in conservative methionine transferring to threonine.His mother also held the mutation,which was not found in 98 control samples.So T6253C was considered the nosogenetic mutation.Conclusion This is the first time to report a mutation in MTCO1 responsible of MELAS.

Objective To investigate the expression levels of platelet microparticles(PMPs)in patients with diabetic nephropathy(DN)and their relationship with renal injury.Methods Thirty DN patients,30 T2DM patients without DN and 30 healthy controls were enrolled.Flow cytometry was used to detect the quantity and phenotype of PMPs,and ELISA was used to measure the levels of plasma renin,angiotensin Ⅱ(angiotoninⅡ,AngⅡ),vascular endothelial growth factor(vascular endothelial growth factor,VEGF)and transforming growth factor-β1(transforming growth factor-β1,TGF-β1).Kidney function was determined by blood biochemistry.Results The quantity of PMPs in the DN group was(1 564±346)particles/μL,which was significantly higher than that in the non-DN group(1 246±312)particles/μL and the control group(1 223±299)particles/μL(P＜0.05).The PMPs in the DN group mainly expressed CD62P and CD41a,accounting for(76.5±12.3)%.Moreover,the levels and activation of PMPs increased with the progression of DN.The quantity of PMPs was positively correlated with renin and other factors(r=0.56-0.62,P＜0.05),negatively correlated with the glo-merular filtration rate(GFR)(r=-0.64,P＜0.05),and positively correlated with the urinary albumin excretion rate(UAE)(r=0.66,P＜0.05).PMPs were also an independent influencing factor of UAE(β=12.34,P＜0.05).Conclusion PMPs expression is elevated in DN patients,which may be associated with renal injury,but it is insufficient to confirm its role in the pathogenesis of DN.

OBJECTIVETo observe the effects of Relinqing granules (powder of Polygonum capitatum extract) on the bacterial pyelonephritis model in rats.

METHODThe rat bacterial pyelonephritis model was induced by injecting the escherichia coli ATCC-25922 into kidney parenchyma. The rats were divided ramdamly into Relinqing groups(52.32, 26.16 g x kg(-1)), norflorin group (0.03 g x kg(-1)), model group and normal control group, and were given experimental drugs by gastrogavage. The contents of leucocytes (WBC), occult bloo (BLD), glucose (GLU), protein (PRO), ketones, bilirubin and urobilinagen in urine were determined.

RESULTAs compared with the model group, Relinqing granules 6.0 g x kg(-1) (crude drug 52.32 g x kg(-1)) could decrease significantly the contents of WBC and BLD in urine and, however, had no markedly effects on the other biochemical parameters of urine.

CONCLUSIONRelinqqing granule has significant effects of decreasing urine WBC and BLD on the bacterial pyolonephritis in rats.

Animals ; Anti-Infective Agents, Urinary ; isolation & purification ; pharmacology ; Bilirubin ; urine ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Escherichia coli Infections ; urine ; Female ; Glycosuria ; urine ; Ketones ; urine ; Leukocyte Count ; Male ; Occult Blood ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Proteinuria ; urine ; Pyelonephritis ; urine ; Rats ; Rats, Sprague-Dawley