Effects of Genipin on the expression of uncoupling protein 1 in brown adipose and white adipose tissues in mice / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of genipin on promoting brown adipose tissue activation and white adipose tissue browning.@*METHODS@#The male C57BL/6J mice were divided into three groups: normal control group, genipin group and cold-stimulus group.Genipin group were treated consecutively with genipin at a dose of 15 mg/kg once a day for 9 days, normal control group were treated with the saline.The mice with cold-stimulus were exposed to 4℃ environment for 5 days.Daily food amount and body weight were measured.Morphological changes were observed in the subscapular region, inguinal region and epididymis around the adipose tissue.The expression of uncoupling protein 1 (UCP1) was determined by real-time PCR and Western blot respectively.@*RESULTS@#The wet weight of white fat in genipin-treated mice was decreased by 16% , and 28% in that of cold-stimulus mice, compared with the normal control group (P＜0.05).After treatments of genipin and cold-stimulus, the color of white adipose tissues was darker, and the size of lipid droplets in adipocytes was smaller, whereas the number was increased.Compared with the normal control group, UCP1 expression was increased obviously in fat tissues, including the subcutaneous and visceral white adipose tissues, and brown adipose tissue after treated with genipin and cold-stimulus (P＜0.05).@*CONCLUSION@#Genipin promoted activation of brown adipose tissue and browning of white adipose tissue by upregulating UCP1 expression, which could contribute to the loss of body weight against obesity.

Cloning and expression analysis of tocopherol cyclase gene in Carthamus tinctorius / 中国中药杂志

The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named CtTC . Bioinformatics analysis showed theopen reading frame （ORF）of CtTC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62.9 kDa and theoretically isoelectric point was 5.01.Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide. The subcellular localization showed that the CtTC protein was located in the chloroplast. The expression of CtTC gene in safflower seeds at different development stages was determined by quantitative real-time PCR, it was found that the highest expression level of CtTC gene was detected in 50 DAF.Quantitative RT-PCR analysis suggested that expression of CtTC is induced and strengthened by drought stresses. This research provided a candidate gene for metabolic engineering of vitamin E and resisting stress.

Failure to Identify Somatic Mutations in Monozygotic Twins Discordant for Schizophrenia by Whole Exome Sequencing / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Schizophrenia (SCZ) is a severe, debilitating, and complex psychiatric disorder with multiple causative factors. An increasing number of studies have determined that rare variations play an important role in its etiology. A somatic mutation is a rare form of genetic variation that occurs at an early stage of embryonic development and is thought to contribute substantially to the development of SCZ. The aim of the study was to explore the novel pathogenic somatic single nucleotide variations (SNVs) and somatic insertions and deletions (indels) of SCZ.</p><p><b>METHODS</b>One Chinese family with a monozygotic (MZ) twin pair discordant for SCZ was included. Whole exome sequencing was performed in the co-twin and their parents. Rigorous filtering processes were conducted to prioritize pathogenic somatic variations, and all identified SNVs and indels were further confirmed by Sanger sequencing.</p><p><b>RESULTS</b>One somatic SNV and two somatic indels were identified after rigorous selection processes. However, none was validated by Sanger sequencing.</p><p><b>CONCLUSIONS</b>This study is not alone in the failure to identify pathogenic somatic variations in MZ twins, suggesting that exonic somatic variations are extremely rare. Further efforts are warranted to explore the potential genetic mechanism of SCZ.</p>

Inspiratory muscle training followed by non-invasive positive pressure ventilation in patients with severe chronic obstructive pulmonary disease: a randomized controlled trial / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of inspiratory muscle training followed by non-invasive positive pressure ventilation in patients with severe chronic obstructive pulmonary disease (COPD).</p><p><b>METHODS</b>This investigator-initiated randomized, controlled trial recruited 88 patients with stable GOLD stage IV COPD, who were randomized into 4 equal groups to continue oxygen therapy (control group) or to receive inspiratory muscle training followed by non-invasive positive pressure ventilation (IMT-NPPV group), inspiratory muscle training only (IMT group), or noninvasive positive pressure ventilation only (NPPV group) for at least 8 weeks. The outcomes of the patients were assessed including the quality of life (SRI scores), maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), dyspnea (MRC scores), 6-min walking distance (6MWD) and lung function.</p><p><b>RESULTS</b>s Compared to baseline values, SRI scores, 6MWT and MRC scores increased significantly after 8 weeks in IMT-NPPV, IMT and NPPV groups, and the improvements were significantly greater in IMT-NPPV group than in IMT and NPPV groups (P<0.05 for all). In IMT-NPPV and IMT groups, MIP and MEP increased significantly after the training (P<0.05), and the improvement was more prominent in IMT-NPPV group (P<0.05). No significant changes were found in pulmonary functions in the groups after 8 weeks of treatment (P>0.05).</p><p><b>CONCLUSION</b>Inspiratory muscle training followed by non-invasive positive pressure ventilation, compared with inspiratory muscle training or non-invasive positive pressure ventilation alone, can better enhance the quality of life, strengthen the respiratory muscles, improve exercise tolerance and relieve the dyspnea in patients with COPD.</p>

Cloning and expression analysis of CtbZIP20 transcription factor gene from Carthamus tinctorius and construction of plant expression vector / 中草药

Objective: To clone bZIP20 (basic region/leucine zipper motif) gene from Carthamus tinctorius, analyze the expression level in different plant tissues, and construct the plant expression vector. Methods: The bZIP20 gene was cloned by RT-PCR techniques, and the protein characteristics were analyzed by bioinformatics, and phylogenetic tree was constructed. The expression of bZIP20 gene in different tissues and the roots after inoculated by Fusarium oxysporum were analyzed using real time-PCR, and the plant expression vector pBASTA-bZIP20 was constructed. Results: The ORF sequence of bZIP20 gene was 981 bp, encoded a protein of 326 amino acids (GenBank: KT692605). Sequence alignment and phylogenetic tree analyses showed that bZIP20 had 85.41% and 83.99% of consistency with bZIP of Sesamum indicum and Camellia assamica. Real-time PCR results showed significant differences, the highest expression level of bZIP20 gene was detected in flower, and was highest in the bud period, bZIP20 gene was significantly increased in root tissue inoculated with F. oxysporum. The plant expression vector pBASTA-bZIP20 was obtained. Conclusion: The bZIP20 gene of safflower is successfully cloned, and the expression is analyzed. The plant expression vector pBASTA-bZIP20 is constructed.

Study on expression of metallothionein 2 gene in safflower / 中草药

Objective: To obtain the transgenic safflower plants which expressed Arabidopsis thaliana metallothionein 2 (MT2) gene, and lay a foundation for development of MT products. Methods: The oleosin-MT gene was obtained from pEASY-oleosin-MT by Nco I/Hind III, then was inserted into plant expression vector pOP. The recombinant plasmid named pOP-oleosin-MT was transferred into Agrobacterium tumefaciens EHA105.The oleosin-MT gene was introduced into safflowers via Agrobacterium-mediated method and positive transgenic plants were determined by PCR analysis. Results: The recombinant plasmid pOP-oleosin-MT was successfully constructed. PCR and Southern blotting analysis confirmed that MT gene was integrated into the genome of safflower plant and three transgenic plants were obtained. Conclusion: The safflower regeneration system is constructed successfully and MT gene is successfully transformed into safflower plant.

Diagnostic Value of (18)F-FDG-PET/CT for Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the diagnostic value of (18)F-FDG-PET/CT in multiple myeloma (MM).</p><p><b>METHODS</b>A total of 66 patients, who were highly suspected of MM in our hospital from July 2012 to December 2014, were chosen as study objects. All patients were diagnosed or excluded by pathological examination. All patients were detected by (18)F-FDG-PET/CT, and its diagnostic value was analyzed. The number of focuses were counted.</p><p><b>RESULTS</b>Out of 66 patients 59 patients (89.39%) were diagnosed with multiple myeloma. The sensitivity of PET was 98.31%, the specificity of PET was 85.71%, the Youden index was 0.8402; the sensitivity of CT was 96.61%, the specificity of CT was 85.71%, the Youden index was 0.8232; the sensitivity of PET/CT was 100.00%, the specificity of PET/CT was 83.33%, the Youden index was 0.8333. In 59 MM patients, 635 focuses were detected, out of them 572 focuses (90.08%) were detected by CT, 593 focuses (93.39%) were detected by PET, 530 focuses (83.46%) were coincided on PET/CT.</p><p><b>CONCLUSION</b>(18)F-FDG-PET/CT has diagnostic value for multiple myeloma, and it can be used in locating and counting focuses, as well as in evaluating the treatment efficacy and guiding the clinical work.</p>

Cloning and prokaryotic expression of transcription factor CtMYB1 gene in Carthamus tinctorius / 中草药

Objective: To obtain a transcription factor gene MYB, clone a CtMYB1 gene from the safflower petals of Carthamus tinctorius, and performe its sequence analysis and prokaryotic expression vector construction. Methods: According to high expression Unigene123993 sequence in safflower transcriptome, MYB gene was cloned from safflower by RT-PCR and RACE methods. The full-length cDNA sequences CtMYB1 gene as templates, the open reading frame (ORF) of cDNA sequences was obtained by PCR. Prokaryotic expression vector pEASY-E1-CtMYB1 was constructed and the expression in E. coli BL21 (DE3) was transformed. Results: A MYB gene was successfully cloned from the safflower petals of C. tinctorius, named CtMYB1 (GenBank accession No. KJ524853). A full length cDNA of CtMYB1 was 893 bp and ORF was 750 bp, encoding a protein of 249 amino acid. The prokaryotic expression vector was obtained. SDS-PAGE results showed that the molecular weight was 30000, same with the relative molecular weight of predicted protein. Conclusion: CtMYB1 is cloned from safflower and the prokaryotic expression vector is constructed, which preliminarily proves that the gene is successfully expressed in E. coli.

MiRNA-365 and miRNA-520c-3p respond to risperidone treatment in first-episode schizophrenia after a 1 year remission / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>MicroRNAs (miRNAs) control gene expression by destabilizing target transcripts and inhibiting their translation. Aberrant expression of miRNAs has been described in many human diseases, including schizophrenia. However, the effects on miRNA expression in response to antipsychotic treatment in peripheral circulation have not been thoroughly examined.</p><p><b>METHODS</b>Using quantitative real-time PCR (qRT-PCR), We quantified the expression of seven candidate miRNAs in plasma samples of 40 first-episode schizophrenics before and after antipsychotic treatment. The patients were all treated with risperidone and achieved remission in 1 year.</p><p><b>RESULTS</b>Compared with the baseline, the expression levels of miR-365 and miR-520c-3p were significantly down-regulated after 1 year of risperidone treatment (P < 0.001). There were no significant correlations between the clinical symptoms and the expression levels of these two miRNAs (P > 0.05).</p><p><b>CONCLUSIONS</b>This study analyzed possible circulating miRNAs in response to antipsychotic monotherapy for schizophrenia, the further mechanism need to be confirmed.</p>

Establishment of the Chang liver cell line stably overexpressing human UCP2 gene and its effect on mitochondrial membrane potential and reactive oxygen species / 中华肝脏病杂志

To establish the Chang liver cell line stably overexpressing human uncoupling protein 2 (UCP2) and observe the effect of UCP2 on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The Chang liver cell line was transfected with recombinant plasmid containing full-length human UCP2 cDNA (pcDNA3.1-hUCP2) or pcDNA3.1 empty vector. The stable cell line was established by antibiotic screening with Zeocin. UCP2 expression was detected by Western blotting and immunocytochemistry. The UCP2 overexpressing cells were pretreated with genipin at various doses (25, 50 and 100 munol/L). MMP and intracellular ROS were detected by fluorescence spectrophotometry. The total normalized protein content in UCP2 overexpressing cells was 1.6-fold higher than that in unmanipulated normal cells. The fluorescence intensities of Rhodamine123 and DCFH-DA in UCP2 overexpressing Chang liver cells (11.11+/-2.76 and 4.97+/-0.62, respectively) were significantly lower than those in unmanipulated normal cells (15.56+/-2.55, P less than 0.01 and 6.14+/-1.25, P less than 0.05, respectively) and in cells transfected with empty vector (16.11+/-2.93, P less than 0.01 and 6.23+/-1.13, P less than 0.05, respectively). Treatment of UCP2 overexpressing cells with 25, 50 and 100 munol/L genipin caused a dose-dependent increase in fluorescence intensities of Rhodamine123 (14.89+/-2.89, 17.89+/-2.93 and 24.00+/-2.55, respectively, all P less than 0.01) and DCFH-DA (9.16+/-0.78, 10.84+/-1.09 and 11.83+/-1.25, respectively, all P less than 0.01). The Chang liver cell line stably overexpressing UCP2 was established successfully. Using this cell system, UCP2 was found to play a role in mitochondrial function by regulating MMP and ROS.

Expression of nucleophosmin in glandular epithelium of non-pregnant human endometrium during the menstrual cycle / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nucleophosmin plays a critical role in embryonic development. This study aimed to examine the expression pattern of nucleophosmin in glandular epithelium of human endometrium during the menstrual cycle.</p><p><b>METHODS</b>Endometrial tissues used for this study were obtained from 46 non-pregnant patients who underwent hysterectomy which had been performed to treat benign diseases. Nucleophosmin expression was assessed by in situ hybridization and immunohistochemistry.</p><p><b>RESULTS</b>At the early-, mid- and late-proliferative phase, nucleophosmin mRNA was highly expressed in glandular epithelium of human endometrium. At the secretory phase, the expression of nucleophosmin mRNA was reduced in glandular epithelium in early-secretory phase, and the expression in mid- and late-secretory phases was not detected. Similarly, nucleophosmin protein was strongly expressed in endometrial glands throughout the proliferative phase, but was gradually reduced during secretory phase.</p><p><b>CONCLUSION</b>Nucleophosmin mRNA and protein are expressed in glandular epithelium of human endometrium throughout the menstrual cycle.</p>

Inhibiting effect of HPV16 E6 small interfering RNA on the growth of human cervical carcinoma xenotransplanted in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the inhibiting effect of human papillomavirus 16 E6-specific small interfering RNA (siRNA) on cervical cancer transplanted in nude mice.</p><p><b>METHODS</b>CaSki cells transfected with HPV16 E6 siRNA were transplanted into nude mice. HPV16 E6 siRNA was injected into the tumor, and the control group was treated with NS. Tumor growth was monitored once every other day. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) was performed to detect apoptosis. The expression of E6 and p53 protein was assessed by immunohistochemistry. The contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured and the liver and kidney were examined by histopathology.</p><p><b>RESULTS</b>Inhibition of tumor growth was demounstrated after treatment with HPV16 E6 siRNA. The volume and weight of tumors were significantly decreased in comparison with those of control group (P < 0.05). Apoptosis occurred more frequently in the experiment group than in the control. The expression of E6 and p53 was down-regulated. The contents of ALT and AST underwent no significant changes, and the histopathology of liver and kidney showed no abnormal changes.</p><p><b>CONCLUSION</b>The growth ability of human cervical cancer xenograft tumors in nude mice can be inhibited by HPV16 E6-specific siRNA, with no toxic side effect on the liver. It may provide an useful method of gene-therapy against cervical cancer.</p>

Protective effects of glucagon-like peptide 2 on intestinal ischemia/reperfusion injury in mice / 中国应用生理学杂志

<p><b>AIM</b>To investigate the protective effects of glucagon-like peptide 2(GLP-2) on intestinal ischemia/reperfusion (I/R) injury in mice.</p><p><b>METHODS</b>Intestinal ischemia/reperfusion model in mice were set up and 32 mice of Kunming species were divided randomly into 4 groups (n=8): Sham group, I/R group, I/R + GLP-2 group and I/R + glutamine group. The morphologic changes of intestinal mucosa were observed under LM. The villus height and crypt depth of intestine, the activity of diamine oxidase (DAO) in intestine and bacterial translocation rates of mesenteric lymph nodes (MLN) were detected.</p><p><b>RESULTS</b>Compared with sham operation group, the intestinal villi were sloughed in I/R group with decreased villus height and crypt depth (P < 0.01), the DAO activities were decreased (P < 0.01), and MLN bacterial translocation rates were increased (P < 0.05). While GLP-2 administration improved the villus damage, increased DAO activity (P < 0.01), and decreased MLN bacterial translocation rates (P < 0.05), compared with I/R group.</p><p><b>CONCLUSION</b>GLP-2 have protective effects on intestinal morphology and barrier function after ischemia/reperfusion injury in mice.</p>

Effects of vagotomy on UCP2 mRNA expression and gastric acid secretion in rats / 中国应用生理学杂志

<p><b>AIM</b>The relationship between gastric acid secretion and ATP level, and regulation of uncoupling protein 2 (UCP2) mRNA expression by vagus nerve were studied in vagotomies rats.</p><p><b>METHODS</b>With the high selective vagotomy model, the gastric acidity was titrated to pH 7.0 with 0.1 mol/L NaOH solution and ATP contents were quantified by using fluorimetry. The expression of UCP2 mRNA was observed by using Northern blot in stomach of rats.</p><p><b>RESULTS</b>Both of gastric acidity and ATP contents in stomach body decreased significantly at 24 h after vagotomy. The expression of UCP2 mRNA was markedly increased as compared with sham operation group.</p><p><b>CONCLUSION</b>ATP contents decreased and vagus nerve down-regulates expression of UCP2 mRNA in stomach corpus in vagotomies rats. The results indicates that vagus nerve could underlay the gastric acidity by inhibiting expression of UCP2 mRNA and increasing ATP contents in rats.</p>