Up- and Down-regulated Leukemia-related Protein 16 Affects ERα Expression and Prolactin Secretion by GH3 Cells / 生物医学与环境科学（英文）

Prolactinoma is an estrogen-related tumor and leukemia-related protein 16 (LRP16) is correlated with the progression of estrogen-related tumors, but the regulatory mechanism between LRP16 and prolactinoma remain unclear. This study demonstrates a variation in LRP16 with estrogen receptor α (ERα) in prolactinoma models and the up and downregulation effects of LRP16 on prolactin secretion of pituitary adenomas cells (GH3 cells). In our study, 50 male SD rats (30-day-old) were randomly divided into five groups of 10 rats each. After 120 days of treatment, the rats were sacrificed, and the expression of LRP16 and ERα were examined by Western blot and immunohistochemistry to explore the changes in ERα, LRP16, and prolactin. After siRNA transfection of the respective genes, the GH3 cells were cultured, and their secretory function as well as the expression of ERα mRNA and prolactin were analyzed by enzyme-linked immunosorbent assay and real-time-polymerase chain reaction analysis. The results show that secretion of prolactin by GH3 cells can be affected by up and downregulating LRP16 expression, which may provide a novel medical therapy in clinical trials.

Gene Expression Profile of Persistent Postoperative Hypertension Patients with Aldosterone-producing Adenomas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hypertension often persists after adrenalectomy for primary aldosteronism (PA). Many studies have analyzed the outcomes of adrenalectomy for aldosterone-producing adenomas (APA) to identify predictive factors for persistent hypertension. However, differentially expressed genes in persistent postoperative hypertension remain unknown. Our aim was to describe gene expression profile of persistent postoperative hypertension patients with APA.</p><p><b>METHODS</b>In this study, we described and compared gene expression profiles in persistent postoperative hypertension and postoperative normotension in Chinese patients with APA using microarray analysis. Confirmation was performed with quantitative real time-polymerase chain reaction analysis. Bioinformatic analysis (gene ontology analysis, pathway analysis and network analysis) was used for further research.</p><p><b>RESULTS</b>Microarray analysis identified a total of 99 differentially expressed genes, including 18 up-regulated and 81 down-regulated genes. Among the dysregulated genes were fat atypical cadherin 1 as well as fatty acid binding protein 4 and other genes that have not been previously studied in persistent postoperative hypertension with APA. Bioinformatics analysis indicated that differentially expressed genes were associated with lipid metabolic process, metal ion binding, and cell differentiation. Pathway analysis determined that five pathways corresponded to the dysregulated transcripts. The mRNAs-ncRNAs co-expression network was composed of 49 network nodes and 72 connections between 18 coding genes and 31 noncoding genes.</p><p><b>CONCLUSIONS</b>This study revealed differentially expressed genes in persistent postoperative hypertension with APA and provided a resource of candidate genes for exploration of possible drug targets and prognostic markers.</p>

Glitazones protects beta cell function from cytotoxic cytokines through PPAR gamma-dependent mechanisms / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of glitazones on islet beta cells and PPAR gamma dependence of such effects.</p><p><b>METHODS</b>IL-1beta and IFN-gamma were used to treat NIT-1 cells, a beta cell line, to induce beta cell damage. The cells were pretreated with rosiglitazone and pioglitazone at different concentrations to study the protective effects of these drugs. The cell apoptosis rate was determined with Annexin V-FITC by flow cytometry, and the insulin secretion capacity of the cells was assessed with ELISA. GW9662 and PPARgamma-SiRNA were used to specifically inhibit PPAR to investigate the PPAR gamma-dependent mechanisms.</p><p><b>RESULTS</b>Rosiglitazone and pioglitazone at 10 micromol/L could significantly decrease the apoptosis of beta cells induced by the cytokines (apoptotic rates of 13.99% and 16.67% vs 51.33%, P<0.01). Rosiglitazone at 10 micromol/L and pioglitazone at 20 micromol/L were less effective than 10 micromol/L rosiglitazone and pioglitazone. The insulin secretion of the cytokine-treated cells decreased from 8.5-/+0.6 ng/ml of the control group to 3.6-/+0.5 ng/ml, while rosiglitazone and pioglitazone could increase the insulin secretion to 6.8-/+0.7 ng/ml and 5.9-/+0.9 ng/ml, respectively. When PPAR gamma was specifically inhibited by GW9662 and PPARgamma-SiRNA, the protective effects of rosiglitazone and pioglitazone were almost undetectable, and the apoptotic rate increased and insulin secretion decreased to the level of the cytokine-treated cells.</p><p><b>CONCLUSION</b>Glitazones can protect beta cells from apoptosis and impairment of insulin secretion function resulting from the cytotoxic cytokines via a PPAR gamma-dependent mechanism.</p>

Analysis on iodized salt surveillance of Changzhou City in Jiangsu Province during 1996-2007 / 中国地方病学杂志

Objective To explore the qualification and consuming of iodized salt at wholesale and household levels after Salt Iodization.Methods Iodized salt surveillance at wholesale and household levels every year by detecting iodine content.Direct titration method(GB/T 13025.7-1999)was used for salt iodjne detecting and arbitration method was used for Sichuan salt and special salt.Results Five thousand six hundred and seventy five samples of 227 batches from 3 wholesale industries were detected during 1996-2000,batch qualification rate was 60.79%(138/227)and iodized salt qualification rate was 61.83%(3509/5675).During 2001-2007,2556 samples of 252 batches from wholesale levels were detected.The batch qualification rate and iodized salt qualification rate were 1 00%(252/252)and 99.88%(2553/2556),respectively.At household level.1583 samples from 236 villages were detected during 1996-2000.Iodized salt qualification rate was 74.24%(1 170/1576)and consuming rate of qualified iodized salt was 73.91%(1 170/1583)and iodine median was 45.14 mg/kg.During 2001-2007,13 140 samples from 1656 villages were detected.Iodized salt qualification rate,consuming rate 0f qualified iodized salt and iodine median were 98.03%(12 830/13 088),97.64%(12 830/13 140)and 30.13 mg/kg,respectively. The most difference of iodine content was 3.46 mg/kg in 3 wholesale industries.At household level there was a 4.95%reduction in comparison with at wholesale level.Conclusions Salt iodization level and edible iodine salt reach the national requirements of iodine deficiency control from the starting stage.The quality 0f iodized saIt at household level related to the exclusive wholesale industry and loss phenomenon maybe existed when salt was sold from wholesale industries to residents.

Construction and identification of recombinant avian adeno-associated virus expressing GFP reporter gene / 病毒学报

To generate recombinant avian adeno-associated virus (rAAAV) for gene transfer studies in avian cells, the recombinant plasmid containing the whole genome of AAAV was digested with restriction enzymes to remove the Rep and Cap genes, resulting in AAAV transfer vector pAITR. GFP-expressing cassette was amplified by PCR and inserted into the AAAV transfer vector. The Rep-Cap gene of AAAV amplified by high fidelity PCR was subcloned into eukaryotic expression vector pcDNA3, resulting in an AAAV helper vector pcDNA-ARC. The Rep and Cap genes amplified by high fidelity PCR were subcloned separately into the co-expression vector pVITRO2-mcs, resulting in another AAAV helper vector pVITRO2-ARC. Using calcium phosphate precipitation method, rAAAV-GFP was generated by co-transfecting AAV-293 cells with a cocktail of pAITR-GFP, pcDNA-ARC or pVITRO2-ARC, and adenovirus helper vector pHelper. The three structural proteins VP1, VP2 and VP3 of correct molecular masses were detected by SDS-PAGE and the GFP reporter gene was detected by PCR in purified rAAAV-GFP virions. Chicken embryonic fibroblast (CEF) cells and CEL cell line were transduced with the recombinant virus, the GFP-positive cells were easily observed under fluorescent microscope, expression of which lasted for at least two weeks. These data demonstrate that an efficient helper virus-free packaging system has been established for generating recombinant AAAV particles for gene transfer studies in avian cells and for development of recombinant vaccines against avian diseases.