Effect of laparoscopic sleeve gastrectomy on sex hormone in male severe obesity / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of laparoscopic sleeve gastrectomy(LSG) on sex hormone in male patients with severe obesity.</p><p><b>METHODS</b>Retrospective analysis was performed in 31 male patient with severe obese [body mass index(BMI) ≥28 kg/m, obesity group] who underwent LSG in Shanghai Tenth People's Hospital of Tongji University from December 2012 to May 2016. The anthropometric parameters(weight, BMI, waist circumference, hip circumference, waist-hip ratio, body fat percentage), glucose metabolic indices [fasting plasma glucose(FPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index(HOMA-IR)], and sex hormone parameters [estradiol(E2), total testosterone (TT), follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] were collected preoperatively and 1, 3, 6 months postoperatively. In addition, 31 healthy male volunteers with normal BMI were consecutively recruited in this study as control group. The above-mentioned parameters were also determined in control group. Changes of these variables before and after surgery were analyzed. Pearson method was used to analyze the correlation of TT with anthropometric parameters and glucose metabolic indices before and after surgery.</p><p><b>RESULTS</b>The average age of patients in obesity and control group was (32.9±9.7) (18 to 56) years and (30.7±8.9) (18 to 49) years. Compared to the control group, obesity group had significantly higher anthropometric parameters and glucose metabolic indices before surgery (all P<0.05). In obesity group, the anthropometric and glucose metabolic indices significantly decreased at 1 to 6 months after surgery compared to those before surgery (all P<0.05). At 1 month after surgery, the anthropometric parameters and glucose metabolic indices in obesity group were significantly higher than those in control group (all P<0.05). At 3, and 6 months after surgery, there were no significant differences in glucose metabolic indices between obesity and control group (all P>0.05), while the anthropometric parameters in obesity group were still significantly higher than those in control group(all P<0.05). The sex hormone parameters in control and obesity group before surgery were as follows: E2: (100.2±23.5) pmol/L and (129.2±81.9) pmol/L; TT: (18.0±4.9) nmol/L and (8.4±4.5) nmol/L; FSH: (4.5±3.1) IU/L and (4.3±2.5) IU/L; LH: (4.4±1.7) IU/L and (5.3±2.6) IU/L. Compared to control group, the TT level of obese patients before surgery significantly decreased(P=0.000), while no significant differences were observed in the levels of E2, FSH, and LH(all P>0.05). The TT levels were significantly increased at 1, 3, 6 months after surgery[(13.1±7.0), (13.6±5.7), (21.0±19.3) nmol/L, respectively, all P<0.05] and the E2 level was significantly decreased at 6 months after surgery [(91.4±44.9) pmol/L, P<0.05], while no significant differences were observed at 1 and 3 months after surgery (all P>0.05). Furthermore, the FSH and LH levels did not exhibit significant change at 1, 3, and 6 months after surgery compared to those before surgery (all P>0.05). At 1 month after surgery, no significant correlations were examined in the change value of TT levels (▹TT) with the changes of BMI(▹BMI), FPG(▹FPG), FINS(▹FINS), HOMA-IR(▹HOMA-IR), and E2(▹E2) (all P>0.05). At 3 months after surgery, ▹TT was negatively correlated with ▹BMI (r=-0.441, P=0.015), ▹FINS (r=-0.375, P=0.041), and ▹HOMA-IR(r=-0.397, P=0.030), but not correlated with ▹FPG and ▹E2 (all P>0.05). At 6 months after surgery, ▹TT was negatively correlated with ▹BMI(r=-0.510, P=0.018) and ▹HOMA-IR (r=-0.435, P=0.049), but not correlated with ▹FPG, ▹FINS and ▹E2 (all P>0.05).</p><p><b>CONCLUSIONS</b>Male severe obese patients are accompanied with abnormal sex hormone levels. LSG has a significant effect on weight loss and blood glucose improvement, and may ameliorate the sex hormone unbalance by improving the insulin resistance in men with severe obesity.</p>

Identifying transcription factors involved in Arabidopsis adventious shoot regeneration by RNA-Seq technology / 生物工程学报

Transcriptional regulation is one of the major regulations in plant adventious shoot regeneration, but the exact mechanism remains unclear. In our study, the RNA-seq technology based on the IlluminaHiSeq 2000 sequencing platform was used to identify differentially expressed transcription factor (TF) encoding genes during callus formation stage and adventious shoot regeneration stage between wild type and adventious shoot formation defective mutant be1-3 and during the transition from dedifferentiation to redifferentiation stage in wildtype WS. Results show that 155 TFs were differentially expressed between be1-3 mutant and wild type during callus formation, of which 97 genes were up-regulated, and 58 genes were down-regulated; and that 68 genes were differentially expressed during redifferentiation stage, with 40 genes up-regulated and 28 genes down-regulated; whereas at the transition stage from dedifferentiation to redifferention in WS wild type explants, a total of 231 differentially expressed TF genes were identified, including 160 up-regualted genes and 71 down-regulated genes. Among these TF genes, the adventious shoot related transcription factor 1 (ART1) gene encoding a MYB-related (v-myb avian myeloblastosis viral oncogene homolog) TF, was up-regulated 3 217 folds, and was the highest up-regulated gene during be1-3 callus formation. Over expression of the ART1 gene caused defects in callus formation and shoot regeneration and inhibited seedling growth, indicating that the ART1 gene is a negative regulator of callus formation and shoot regeneration. This work not only enriches our knowledge about the transcriptional regulation mechanism of adventious shoot regeneration, but also provides valuable information on candidate TF genes associated with adventious shoot regeneration for future research.

Study on clinical characteristics of metabolic-based obesity classification / 中华内分泌代谢杂志

Objective This study aimed to explore clinical characteristics of four types of obesity based on metabolic classification. Methods Forty-eight obese patients were divided according to their clinical characteristics into 4 groups including metabolic healthy obesity (MHO), hypometabolic obesity (LMO), hypermetabolic obesity (HMO), and metabolic obesity with inflammation (IMO). 20 normal weight individuals were also recruited as a control group. Body fat, body weight, visceral index, and basal metabolism were measured by Omron body fat meter. Fat content and its distribution were measured by dual energy X-ray absorptiometry. All participating patients underwent various tests for 75 g oral glucose tolerance, blood glucose, insulin, C peptide. Lipid profile, thyroid function and sex hormones levels, and inflammation factors were also measured. Results (1)Patients in MHO group had higher body fat content, but had no metabolic disorder and inflammation. Their hormones levels were normal. (2) Lower metabolic rate and lower hormones levels were found in the patients in LMO group with increasing visceral fat. Trunk/subcutaneous fat mass was significantly higher than that in MHO group(1. 19 ± 0. 25 vs 0. 97 ± 0. 32, P<0. 05). There were abnormal lipid and glucose metabolism in LMO group. The insulin action index was significantly lower than that in MHO group(0. 006 6 ± 0. 002 7 vs 0. 012 1 ± 0. 009 5, P<0. 05). The area under the curve of glucoseconcentrationwassignificantlyhigherinLMOgroupthanthatinMHOgroup[(18.71±8.68vs12.70±4.63) mmol/L, P<0. 05]. (3)Heart rate and blood pressure were higher in HMO group. The heart rate was significantly increased compared with that in MHO group [(90. 50 ± 8. 24 vs 73. 20 ± 14. 11) beat/min, P<0. 05]. The waist circumference was significantly larger than that in MHO group [(111. 88 ± 10. 54 vs 98. 05 ± 15. 56) cm, P<0. 05]. (4) In IMO group, insulin action index was significantly lower than MHO group (0. 007 0 ± 0. 003 3 vs 0.0121±0.0095,P<0.05). ThetrunkfatmassanduricacidlevelsweresignificantlyhigherthanMHOgroup [(17236.38±4610.60vs15816.10±5453.42)gand(468.28±121.32vs376.84±97.14) μmol/L,bothP<0. 05]. Patients in IMO group had acanthosis nigricans, but their glucose level was relatively normal. Conclusion The metabolic-based obese diagnosis is essential for understanding the obesity etiology and providing individualized treatment.

A method for PCR product cloning based on exonuclease III / 生物工程学报

Gene cloning is one of the most important and widely used technologies in molecular biology research. Generally, DNA fragment is cut with restriction enzyme, and then the product is ligated to a linearized vector with complementary sticky end or blunt end by DNA-ligase. This traditional DNA cloning method requires compatible enzyme recognition sites existing in both PCR fragment and targeted vector. Several ligase-free methods have been established to avoid the using of restriction enzyme. However, those methods are time-consuming, labor-intensive and expensive. To overcome these shortcomings, we developed an Exonuclease III based DNA cloning method that takes only 30 minutes with high cloning efficiency and significant economic advantage. Therefore, this method is suitable for large-scale gene cloning.

Digital gene expression profiling analysis of the early adventitious shoot formation in Arabidopsis thaliana / 生物工程学报

Most current research in the field of adventitious shoot formation is focused on the regulatory function of a single gene. However, a systematic transcriptomic analysis of the early adventitious shoot formation is still lacking. Here, we analyzed the transcriptome profiling of the early adventitious shoot formation in Arabidopsis by RNA-seq high throughput sequencing technology, and identified 2 457 differentially expressed genes. Detailed categorization revealed that these genes were mainly involved in hormone homeostasis or signal transduction, callus and lateral root formation, shoot apical meristem development and photosynthesis. Further pathway enrichment analysis showed that genes involved in phenylalanine metabolism and phenylpropanoid biosynthesis were significantly enriched. Moreover, exogenous phenylalanine could repress adventitious shoot formation, indicating that phenylalanine metabolism and phenylpropanoid biosynthesis might be important for adventitious shoot formation.

Identification and expression analysis of WRKY transcription factors in medicinal plant Catharanthus roseus / 生物工程学报

WRKY transcription factors, one of the largest families of transcriptional regulators in plants, involve in multiple life activities including plant growth and development as well as stress responses. However, little is known about the types and functions of WRKY transcription factors in Catharanthus roseus, an important medicinal plant. In this study, we identified 47 CrWRKY transcriptional factors from 26 009 proteins in Catharanthus roseus, and classified them into three distinct groups (G1, G2 and G3) according to the structure of WRKY domain and evolution of the protein family. The expression profiling showed that these CrWRKY genes expressed in a tissue/organ specific manner. The 47 CrWRKY genes were clustered into three types of expression patterns. The first type includes the CrWRKYs highly expressed in flowers and the protoplast treated with methy jasmonate (MeJA) or yeast extraction (YE). The second type contains the CrWRKYs highly expressed in stem and hairy root. The third type represents the CrWRKYs highly expressed in root, stem, leaf, seedling and the hairy root treated by MeJA. Real time quantitative PCR was employed to further identify the expression patterns of the 16 selected CrWRKY genes in various organs, the MeJA-treated protoplasts and hairy roots of Catharanthus roseus, and similar results were obtained. Notably, the expresion of more than 1/3 CrWRKY genes were regulated by MeJA or YE, indicating that these CrWRKYs are likely involed in the signalling webs which modulate the biosynthesis of terpenoid indole alkaloid and plant responses to various stresses. The present results provide a framework for functional identification of the CrWRKYs and understanding of the regulation network of terpenoid indole alkaloid biosynthesis in Catharanthus roseus.

Regulatory networks of somatic embryogenesis in plant / 生物工程学报

The somatic embryogenesis in plant is a very complicated and highly ordered process, which is regulated by many internal and external factors. Among them, gene expression and regulation are key factors. Genes encoding regulatory proteins, for example PLANT GROWTH ACTIVATOR, LEAFY COTYLEDON, BABY BOOM, SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE and PICKLE, interact with each other and form a complicated regulatory network. Recent progress on this regulatory network was reviewed on the basis of our study on the PLANT GROWTH ACTIVATOR 37 gene. In addition, future research perspectives on plant somatic embryogenesis were discussed.