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Short hairpin RNA targeting N-acetylglucosaminyltransferase V inhibits the proliferation of prostate cancer PC-3 cell line.

Yong-hua YAO ; Ting WEI ; Ya-hui LI ; Jian ZHANG

Journal of Southern Medical University.2010;30(6):1253-1255.

OBJECTIVETo investigate the effect of short hairpin RNA (shRNA) targeting N-acetylglucosaminyltransferase V (GnT-V) on the proliferation of prostate cancer PC-3 cells.

METHODSLipofectamine 2000 was used to transfect the recombinant plasmids carrying the shRNA targeting GnT-V gene into PC-3 cells. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to measure the mRNA expression of GnT-V, and CCK-8 assay was used to measure the cell proliferation after the transfection.

RESULTSThe recombinant plasmids were successfully transfected into PC-3 cells, resulting in a reduction of GnT-V mRNA expression by 73%. The proliferation of PC-3 cells was significantly inhibited after the transfection.

CONCLUSIONThe shRNA targeting GnT-V gene can reduce the expression of GnT-V mRNA and inhibit the proliferation of PC-3 cells in vitro.

Cell Line, Tumor ; Cell Proliferation ; Humans ; Male ; N-Acetylglucosaminyltransferases ; genetics ; Prostatic Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

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Mechanisms of foam cell formation in smooth muscle cell derived from bone marrow stem cells.

Peng-ke YAN ; Shi-huang LI ; Cai-wen DUANG ; Zheng-Rong MEI ; Xiang-ping TAN ; Tu-bing SI ; Guo-hong XIAO

Journal of Southern Medical University.2010;30(6):1250-1252.

OBJECTIVETo establish a model of smooth muscle cells differentiated from bone mesenchymal stem cells (BMSC-SMCs) in vitro and explore the relationship between scavenger receptors A (SR-A) and caveolin-1.

METHODSBMSCs were isolated from the femoral bone of SD rats by adherent culture. After treatment of the BMSC-SMCs with 80 mg/L ox-LDL for 72 h, Western blotting was performed to detect the expression of scavenger receptor SR-A, cell cholesterol transport protein ATP-binding cassette transporter Al (ABCA1) and caveolin-1.

RESULTSBMCS-SMCs became foam cells after treatment with ox-LDL. BMSC-SMC gave rise to more foam cell formation than VSMCs did. Western blotting showed that treatment with 80 mg/L ox-LDL for 72 h resulted in significantly increased expression of SR-A and significantly decreased expressions of ABCA1 and caveolin-1.

CONCLUSIONSTreatment of BMCS-SMCs with ox-LDL results in cholesterol ester accumulation in the cells to result in foam cells, the mechanism of which involves up-regulation of scavenger receptor SR-A expression and down-regulation of the reverse cholesterol transport protein ABCA1 and caveolin-1 expression.

ATP Binding Cassette Transporter 1 ; ATP-Binding Cassette Transporters ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; Caveolin 1 ; metabolism ; Cell Differentiation ; Cells, Cultured ; Female ; Foam Cells ; cytology ; Lipoproteins, LDL ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scavenger Receptors, Class A ; metabolism

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Relation between insulin resistance and glutamic acid decarboxylase antibody titers in latent autoimmune diabetes in adults.

Xia LI ; Gan HUANG ; Xiang YAN ; Lin YANG ; Zhi-guang ZHOU

Journal of Southern Medical University.2010;30(6):1247-1249.

OBJECTIVETo investigate the relation between insulin resistance and glutamic acid decarboxylase antibody (GAD-Ab) titers in latent autoimmune diabetes in adults (LADA).

METHODSThe patients with phenotypic type 2 diabetes were screened for GAD-Ab positivity, and the 141 positive patients were divided into two subgroups according to the GAD-Ab titer, namely the high-titer group (LADA-1 subtype) and low-titer group (LADA-2 subgroup). The clinical features and insulin resistance were compared between the two groups. Insulin resistance was calculated by HOMA 2 software, and GAD-Ab and C peptide were determined with radioligand and radioimmune assay, respectively.

RESULTSCompared with low-titer LADA patients, the patients with high titers had younger age of onset, lower BMI, higher HbA1c level, and worse fasting and postprandial C peptide levels. The insulin resistance index by HOMA 2 was significantly lower in LADA-1 group than in LADA-2 group (1.6-/+1.1 vs 2.1-/+1.1, P=0.001). The HOMA2-IR index showed a negative correlation to GAD-Ab titer.

CONCLUSIONThe degree of insulin resistance is correlated to GAD-Ab titers in LADA, and low titer patients have higher insulin resistance level.

Adult ; Aged ; Autoantibodies ; blood ; Autoimmune Diseases ; diagnosis ; immunology ; Diabetes Mellitus, Type 2 ; diagnosis ; immunology ; Female ; Glutamate Decarboxylase ; immunology ; Humans ; Insulin Resistance ; Islets of Langerhans ; immunology ; physiology ; Male ; Middle Aged

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Changes of neurotransmitter endothelin, thromboxance B2 and 6-keto-prostaglandin F1 alpha in patients with chronic pulmonary heart disease.

Yong CHENG ; Zhi-Kui LI

Journal of Southern Medical University.2010;30(6):1244-1246.

OBJECTIVETo explore the changes of the neurotransmitters in patients with chronic pulmonary heart disease (CPHD) and its clinical significance.

METHODSSeventy-two patients with CPHD (42 males, 30 females, mean age 55.6-/+8.9 years) were enrolled in the study, including 48 patients with compensated CPHD and 24 with uncompensated CPHD. Plasma endothelin (ET), thromboxance B2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-K-PGFlalpha) were detected by radioimmunoassay. Thirty blood donors were selected as the normal control.

RESULTSCompared with the normal controls, CPHD patients showed abnormal pulmonary function, and significantly elevated levels of plasma ET and TXB2 (P<0.01) and lowered 6-K-PGFlalpha(P<0.01), but no significant differences were found between the patients with compensated CPHD and uncompensated CPHD (P>0.05). Plasma ET and TXB2 levels were inversely correlated to 6-K-PGFlalpha level (r=-0.4571, P<0.05).

CONCLUSIONThe patients with CPHD present with obvious changes of plasma ET, TXB2 and 6-K-PGFlalpha.

6-Ketoprostaglandin F1 alpha ; blood ; Adult ; Case-Control Studies ; Chronic Disease ; Endothelins ; blood ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Heart Disease ; blood ; Thromboxane B2 ; blood

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Expression, purification and bioactivity evaluation of streptavidin-tagged human interleukin-21 fusion protein.

Ping-ping FA ; Zhen ZHANG ; Jin-long LI ; Zhi-ming HU ; Ji-min GAO

Journal of Southern Medical University.2010;30(6):1240-1249.

OBJECTIVETo obtain streptavidin-tagged human interleukin-21 (hIL21) fusion protein and evaluate its bioactivities.

METHODShIL21-SA-pET21 and pET24a-SA- hIL21 plasmids were constructed and expressed in BL21(DE3) host bacteria. The hIL21-SA and SA- hIL21 fusion protein were purified through Ni-NTA affinity chromatography and refolded by dialysis. Flow cytometry was used to detect hIL21-SA and SA- hIL21 fusion protein on the biotinylated MB49 tumor cells. MTT assay was used to evaluate the effect of the fusion protein on the proliferation of human peripheral blood lymphocytes (PBLs) stimulated by Anti-CD3.

RESULTSThe recombinant fusion proteins were highly expressed in BL21(DE3) at about 30% of the total bacterial proteins. The two fusion proteins exhibited bifunctional activities, i.e. both biotin-binding property and hIL21 activity and SA-mediated high-affinity binding to biotinylated cell surfaces (with anchoring modified rate of about 95.18% and 96.91%).

CONCLUSIONWe have successfully obtained bifunctional fusion protein hIL21-SA and SA- hIL21,which will provide a basis for further study of tumor biotherapy using the proteins.

Cancer Vaccines ; immunology ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Humans ; Interleukins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Streptavidin ; biosynthesis ; genetics

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Cerebral blood flow, cerebral blood volume, oxygen utilization and oxygen extraction fraction: the influence of age.

Yu ZHANG ; Ying-yin PENG ; Guo-yue CHEN ; Wu-fan CHEN

Journal of Southern Medical University.2010;30(6):1237-1239.

OBJECTIVETo analyze the changes of cerebral blood flow (CBF), cerebral blood volume (CBV), oxygen utilization (CMRO2) and oxygen extraction fraction (OEF) with age.

METHODSThe PET images of 7 young (21.0-/+1 years old) and 7 aged volunteers (60.9-/+4.7 years old) were analyzed to identify the areas where CBF, CBV, CMRO2, OEF had significant differences with age. The images were anatomically normalized by statistical parametric mapping (SPM2). A voxel by voxel calculation was performed to obtain the slope with age. Voxels which had statistically significant differences (P<0.05) with age were shown both on global and ROIs brain images.

RESULTSThe CBF decreased with age as was consistent with previous reports. The age-related changes in CBV and CMRO2 were similar to CBF, but OEF increased with age.

CONCLUSIONCBF, CBV and CMRO2 generally decline with age. The increase in OEF with age suggests a greater reduction in CBF than in CMRO2. The most significant decreases of CBF and CMRO2 occur in the convexity of the frontal cortex and inferior parietal cortex in all the functional images, while in the white matter, the influence of age is minimal.

Age Factors ; Aged ; Aging ; metabolism ; physiology ; Blood Volume ; physiology ; Brain ; blood supply ; diagnostic imaging ; Cerebrovascular Circulation ; physiology ; Female ; Humans ; Male ; Middle Aged ; Oxygen Consumption ; physiology ; Positron-Emission Tomography ; methods ; Reference Values ; Regional Blood Flow ; Young Adult

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Clinical features of panereatic disease-associated portal hypertension.

Li-shu XU ; Jian-hu LIU ; Ping LIN ; Kai-hong HUANG ; Qi-kui CHEN ; Yin-ting CHEN ; Zhao-hua ZHU

Journal of Southern Medical University.2010;30(6):1234-1236.

OBJECTIVETo investigate the clinical features and management of pancreatic disease-associated portal hypertension.

METHODSA retrospective analysis was carried out in patients with portal hypertension and concurrent pancreatic diseases. The medical records of these patients were reviewed including the data of demographics, etiologies, venous involvement, clinical presentations, laboratory tests, imaging studies, therapeutic modalities and outcomes.

RESULTSFifty-two patients with portal hypertension resulting from pancreatic diseases were found in our hospital, accounting for 4% of all the patients with portal hypertension in 11 years. The underlying pancreatic diseases were chronic pancreatitis (21 cases, 35.6%), pancreatic carcinoma (20 cases, 33.9%), acute pancreatitis (8 cases, 13.6%), pancreatic pseudocyst (3 cases, 5.1%). Of the 40 patients whose venous involvement was identified, splenic vein obstruction occurred in 27 cases (67.5%) and portal vein obstruction in 16 cases (40.0%). Mild or moderate splenomegaly was present in 48 cases (81.4%), with leukocytopenia as the most common manifestation of the 31 cases (52.5%) with concomitant hypersplenism. Forty-five patients (76.3%) developed gastroesophageal varices (including 35 with isolated gastricvarices), and among them 22 experienced bleeding (42.3%). Conservative treatment was effective in controlling acute bleeding, but could not prevent re-bleeding. Splenectomy was performed in 18 patients mainly due to gastrointestinal hemorrhage. No postoperative bleeding occurred during the follow-up ranging from 8 months to 9 years.

CONCLUSIONPancreatic diseases may compromise portal vein and its tributaries, leading to generalized or regional portal hypertension. Pharmacological therapy can effectively control acute variceal bleeding, while surgical treatment is the appropriate procedure of choice in case of hemorrhagic recurrence.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Esophageal and Gastric Varices ; etiology ; surgery ; Female ; Humans ; Hypertension, Portal ; etiology ; Male ; Middle Aged ; Pancreatic Neoplasms ; complications ; Pancreatitis, Chronic ; complications ; Retrospective Studies ; Young Adult

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Forward genetic screening for zebrafish mutants defective in myelopoiesis.

Zhao-xia DAI ; Guang YAN ; Ying-hua CHEN ; Wei LIU ; Zhong-jun HUO ; Zong-hua WEN ; Jing LIU ; Kun WANG ; Zhi-bing HUANG ; Ning MA ; Xiao-hui CHEN ; Ping-yun MA ; Wei-hao LUO ; Ying ZHAO ; Shu FAN ; Hong-hui HUANG ; Zi-long WEN ; Wen-qing ZHANG

Journal of Southern Medical University.2010;30(6):1230-1233.

OBJECTIVETo identify zebrafish mutants with myelopoiesis defects by ENU mutagenesis and large-scale forward genetic screening.

METHODSMale zebrafish were mutagenized with N-ethyl N-nitrosourea to induce mutations in the spermatogonial cells to generate the founders, which were outcrossed with AB to raise F1 fish. The F1 fish from different founders were mated to generate the F2 families. The F3 embryos from F2 sibling crosses were screened by Sudan black B staining and neutral red staining.

RESULTSA total of 350 F2 families from F1 sibling crosses were screened, and 1424 F2 crosses were analyzed. Six mutations were identified resulting in abnormal Sudan black B staining and neutral red staining, indicating the involvement of neutrophil deficiency or macrophage abnormalities.

CONCLUSIONIt is simple and cheap to induce and screen myelopoiesis deficiency in zebrafish by ENU chemical mutagenesis and Sudan black B staining and neutral red staining. These mutants shed light on the identification of the genes important to myelopoiesis in zebrafish.

Animals ; Gene Expression Regulation, Developmental ; genetics ; Genetic Testing ; Male ; Mutagenesis ; Mutation ; Myeloid Progenitor Cells ; physiology ; Myelopoiesis ; genetics ; Zebrafish ; genetics

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Effects of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells: a study using chromatin immunoprecipitation.

Jian-feng CHEN ; Xin-hua QIAN ; Dan-hua ZHAO ; Xin-lai QIAN

Journal of Southern Medical University.2010;30(6):1222-1225.

OBJECTIVETo develop a real-time PCR-based chromatin immunoprecipitation (ChIP) assay for determining the effect of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells.

METHODSK562 cells were grown in the presence or absence of 0.5 mmol/L sodium butyrate for 48 h, and 1=10(7) cells per group were used for real-time PCR-based ChIP with anti-acetylated histone H3 or H4 antibodies. The levels of acetylated histone H3 and H4 (acH3 and acH4) in Ggamma- and Agamma-globin gene promoter regions were measured.

RESULTSIn the K562 cells with sodium butyrate treatment or without any treatment, the levels of acH3 or acH4 in Ggamma- or Agamma-globin gene promoter were higher than that in the necdin gene (negative control). Compared with the untreated K562 cells, the cells treated with 0.5 mmol/L sodium butyrate showed a 3.1-fold or 2.6-fold increase in acH3 or acH4 in Ggamma-globin gene promoter region, with also a 3.7-fold or 3.2-fold increase in acH3 or acH4 in Agamma-globin gene promoter region, respectively (P<0.01).

CONCLUSIONWe have successfully developed a real-time PCR-based ChIP assay for analyzing the acetylation of histone H3 and H4 in gamma-globin gene promoter regions. Our results support the role of sodium butyrate in increasing the level of acetylated histone in gamma-globin gene promoter regions.

Acetylation ; Butyrates ; pharmacology ; Chromatin Immunoprecipitation ; methods ; Histones ; chemistry ; Humans ; K562 Cells ; Promoter Regions, Genetic ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; gamma-Globins ; genetics

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Effects of exercise on expression and phosphorylation of PI3K and PKB in insulin signaling in the skeletal muscles of type 2 diabetic rats.

Shi-cheng CAO ; Gang ZHAO ; Bo CHANG ; He ZHANG

Journal of Southern Medical University.2010;30(6):1217-1221.

OBJECTIVETo investigate the effect of exercise on the expressions of phosphatidylinositol 3 kinase (PI3K) and protein kinase B (PKB) phosphorylation, protein and glucose transport proteins (GLUT4) at both the protein and mRNA levels in the skeletal muscles of type 2 diabetic rats.

METHODSThirty male SD rats were randomly divided into control group with normal diet feeding, diabetic group and diabetic exercise group with high-fat diet feeding. After 8 weeks of the high-fat diet, each rat received an intraperitoneal injection of streptozotocin (STZ, 30 mg/kg). Three weeks after the injection, the rats were rated for the presence of diabetes, and the rats in the exercise groups took swimming training for 4 weeks; all the groups maintained their assigned diets. The gastrocnemius of all the rats were dissected 48 h after the last training session. Western blotting was applied to detect the phosphorylation and protein expression of PI3K and PKB and the protein expression of GLUT4. The expression of GLUT4 mRNA was determined by RT-PCR.

RESULTSCompared with the diabetic group, the diabetic rats in the exercise group showed significantly increased protein expression and phosphorylation of PKB (P<0.05) and elevated GLUT4 protein and mRNA expressions in the skeletal muscles (P<0.01, P<0.05).

CONCLUSIONExercise training can modulates insulin signal transduction through the protein expression and phosphorylation of the protein kinase to promote glucose uptake in the skeletal muscle of type 2 diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Insulin ; metabolism ; Male ; Muscle, Skeletal ; metabolism ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Phosphorylation ; Physical Conditioning, Animal ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

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