Objective To investigate the effect of testosterone on mitotic orientation in rat prostate epithelial cells and the relative differential gene expression.Methods Twenty SPF male SD rats were divided into 2 groups at random and then subjected to castration.One group of rats was administrated with testosterone 3.7 mg daily for 30 days and the control group was only injected with olive oil.Microscopic analysis was performed using immunohistochemistry.Differential gene expression analysis was conducted by gene microarray and RT-PCR techniques.Results In the testosterone-adminis-trated group, there was a significant mitosis orientation parallel to the basement membrane.But in the control group, mito-sis orientation was oriented perpendicular to the basement membrane.Using the gene microarray and RT-PCR techniques, the cell proliferation genes such as Ran, Tgm4 and Wnt2 in Wnt signal pathway were up-regulated in the testosterone group.Conversely, suppressor cell proliferation genes such as Dkk3 and Fas were down-regulated.Conclusions Mitotic orientation of prostate epithelia cells is changed after testosterone administration.Wnt signal pathway and AR singling path-way also have an influence on the mitosis orientation and cell proliferation.

Adult ; Diagnosis, Differential ; Female ; Humans ; Skin ; pathology ; Skin Neoplasms ; pathology ; Skin Ulcer ; pathology ; Tibia

Hyperlipidemia is a major factor causing coronary heart disease and atherosclerosis. The high-density lipoprotein cholesterol (HDL-C) is a major indicator for measuring lipid levels. However, there is no an effective medicine that can obviously increase HDL-C at present. According to previous laboratory studies, atractylodes macrocephalae extracts could significantly increase HDL-C level. In this study, the metabolic hyperlipidemia rat model was established by feeding high-sugar and fat diets and alcohol-drinking to explore the effect and mechanism of atractylodes macrocephalae extracts on hyperlipidemia rats. According to the findingins, different doses of atractylodes macrocephalae extracts could reduce the levels of TC, TG, LDL-C, ACAT and increase the contents of LCAT, HDL-C. Particularly, the atractylodes macrocephalae extracts (100 mg · kg(-1) group showed increase in HDL-C by about 50% and significant declines in HMG-CoA reductase, TC, TG. In conclusion, Atractylodes Macrocephelae Rhizoma extracts could effectively regulate the dyslipidemia of hyperlipidemia rats, especially on HDL-C. Its mechanism may be related to reduction in cholesterol synthesis by inhibiting HMG-CoA reductase in livers and increase in lipid metabolism and transport by regulating LCAT and ACAT levels.
Acyl Coenzyme A ; antagonists & inhibitors ; genetics ; metabolism ; Animals ; Atractylodes ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; administration & dosage ; Hyperlipidemias ; drug therapy ; enzymology ; metabolism ; Lipoproteins, HDL ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Triglycerides ; metabolism

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

The Rhei Radix et Rhizoma was one of the most widely used traditional Chinese medicine for its special biological activities. The content of rhein, one of its major compounds, was an important standard for the quantity control of Rhei Radix et Rhizoma. The major method used for the detection of rhein was instrumental analysis like HPLC, but it was complex, time-consuming and cannot detect large samples at the same time. The enzyme-linked imunmosorbent assay (ELISA) was accurate, reliable, simple, low costs, and of a high-throughout. Recently, it was widely used for the determination of those small molecule compounds in some traditional Chinese medicinal plants. In this study, an artificial antigen were synthesized by the carbodiimide (CDI) method. Rhein-bovine (rhein-BSA) conju gate and rhein-ovalbumin (rhein-OVA) conjugate, were produced as the immunogen and coating antigen, respectively. The conjugate and the hapten number in the conjugate were determined by UV-Vis spectrophotometry (UV). The conjugation ratio of Rhein and BSA was about 4.0:1, rhein acid and OVA was 2.6 : 1, respectively. Rhein-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The antiserum titer of the Rhein were higher than 8000 detected by ELISA. The successfully synthesized conjugate antigen rhein-BSA implies its feasibility in the establishment of fast immunoassay for the rhein content determination.
Animals ; Anthraquinones ; analysis ; immunology ; Antibodies ; analysis ; immunology ; Antigens, Plant ; analysis ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Mice ; Mice, Inbred BALB C ; Rheum ; chemistry ; immunology ; Rhizome ; chemistry ; immunology

OBJECTIVETo analyze the complications of percutaneous kyphoplasty except bone leakge for the treatment of osteoporotic thoracolumbar vertebral compression fractures.

METHODSFrom October 2008 to October 2012,178 patients with 224 osteoporotic vertebral compression fractures were treated with percutaneous kyphoplasty under local anethsia. There were 72 males and 106 females,ranging in age from 58 to 92 years old,with an average of 75.3 years,including 93 thoracic vertebrae and 131 lumbar vertebrae. The complications except bone cement leakage were analyzed during operation and after operation.

RESULTSAll operations were successful and all patients were followed up from 12 to 60 months with an average of 26.2 months. No death was found. Bone cement leakage occurred in 27 cases, about 15.1% in 178 cases; and complications except bone cement leakage occurred in 15 cases. There was 1 case with cardiac arrest,was completely recovery by cardiopulmonary resuscitation (CPR) immediately; and 1 case with temporary absence of breathing,was recovery after treatment. There were 3 cases with fall of blood pressure and slower of heart rate; 1 case with intestinal obstruction; 2 cases with local hematoma and 1 case with intercostal neuralgia. Vertebral body fractures of 2 cases were split by bone cement and the fractures of adjacent body occurred in 4 cases.

CONCLUSIONIt's uncommon complication except bone cement leakge in treatment of osteoporotic thoracolumbar vertebral compression fractures with percutaneous kyphoplasty. The complication of cardiopulmonary system is a high risk in surgery; and cytotoxicity of bone cement,nervous reflex,fat embolism and alteration of intravertebral pressure may be main reasons.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; surgery ; Humans ; Kyphoplasty ; adverse effects ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery ; Postoperative Complications ; etiology ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

In order to find the cardiotonic constituents of lateral roots of Aconitum carmichaelii Debx., the investigation was carried out. Silica gel column chromatography, Sephadex LH-20, medium-pressure MCI and reverse phase ODS column chromatography were used to separate the 90% EtOH extract of the lateral roots of Aconitum carmichaelii Debx. The structures of the isolated compounds have been identified by chemical properties and spectroscopic analyses. Ten compounds were isolated and their structures were elucidated as benzoic acid-5-hydroxy-2-benzoyl-amino methyl ester (1), honokiol (2), pinoresinol (3), salicylic acid (4), p-hydroxy-cinnamic acid (5), songorine (6), karakoline (7), mesaconitine (8), hypaconitine (9) and 14-benzoylhypaconitine (10), separetely. Compound 1 is a new compound and its structure has been established by NMR, HR-ESI-MS, UV, IR and X-Ray. Compound 2-5 are isolated from the lateral roots of Aconitum carmichaelii Debx. for the first time.
Aconitum ; chemistry ; Cardiotonic Agents ; chemistry ; isolation & purification ; Plant Roots ; chemistry

BACKGROUNDThe CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.

METHODSA SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.

RESULTSOne SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.

CONCLUSIONSWe conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.

Animals ; Chimera ; Genes, env ; HIV-1 ; genetics ; physiology ; Humans ; Macaca mulatta ; Proviruses ; genetics ; Receptors, CCR5 ; physiology ; Simian Immunodeficiency Virus ; genetics ; physiology

OBJECTIVETo evaluate the efficacy of infliximab combined with surgery in the treatment of perianal fistulizing Crohn disease (CD).

METHODSClinical data of 15 patients with perianal fistulizing CD receiving infliximab combined with surgery in the Affiliated Hospital of Nanjing University of Chinese Medicine from March 2010 to June 2011 were analyzed retrospectively. One week after operation, all the patients received infliximab infusion thrice at weeks 0, 2, and 6. Crohn disease activity index (CDAI), perianal Crohn disease activity index (PDAI), body mass index (BMI), routine blood test and endoscopy were evaluated at week 0, 14. Adverse reactions and healing time were recorded.

RESULTSAt week 14, the response rate was 100% with 86.7% (13/15) complete responders. One patient had local improvement and one developed recurrent fistula. The mean healing time was 32.5 (20-45) d. Anorectal stenosis in 4 patients was significantly improved. At week 14, CDAI decreased to 114.0±90.3 from 230.5±97.5 after IFX treatment. PCDAI decreased to 2.8±3.2 from 9.9±3.4, and BMI increased to (21.5±3.0)kg/m(2) from (19.1±3.1)kg/m(2). C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), platelet and neutrophil were significantly decreased from baseline (all P<0.01). Intestinal mucosa healed completely in one patient. There were no serious adverse events except hypokalemia in one patient and severe infusion reaction in another.

CONCLUSIONInfliximab combined with surgery is effective and safe for perianal fistulizing CD.

Adolescent ; Adult ; Antibodies, Monoclonal ; therapeutic use ; Combined Modality Therapy ; Crohn Disease ; complications ; drug therapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Infliximab ; Male ; Rectal Fistula ; drug therapy ; etiology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult