Intestinal microbiota changes in Graves' disease: a prospective clinical study.

Graves' disease (GD) occurs due to an autoimmune dysfunction of thyroid gland cells, leading to manifestations consistent with hyperthyroidism. Various studies have confirmed the link between autoimmune conditions and changes in the composition of intestinal microbial organisms. However, few studies have assessed the relationship between the GD and the changes in intestinal microbiota. Therefore, the present study aimed to investigate changes in intestinal flora that may occur in the setting of GD. Thirty-nine patients with GD and 17 healthy controls were enrolled for fecal sample collection. 16S rRNA sequencing was used to analyze the diversity and composition of the intestinal microbiota. High-throughput sequencing of 16S rRNA genes of intestinal flora was performed on Illumina Hiseq2500 platform. Comparing to healthy individuals, the number of Bacilli, Lactobacillales, Prevotella, Megamonas and Veillonella strains were increased, whereas the number of Ruminococcus, Rikenellaceae and Alistipes strains were decreased among patients with GD. Furthermore, patients with GD showed a decrease in intestinal microbial diversity. Therefore, it indicates that the diversity of microbial strains is significantly reduced in GD patients, and patients with GD will undergo significant changes in intestinal microbiota, by comparing the intestinal flora of GD and healthy controls. These conclusions are expected to provide a preliminary reference for further researches on the interaction mechanism between intestinal flora and GD.

[Effect of Gastrodiae elata-Phallus impudicus sequential planting pattern on soil microbial community structure].

Gastrodia elata is a heterotrophic plant that needed to be symbiotic with Armillaria. The obstacle of continuous cropping in G. elata is serious during the G. elata cultivation, and the mechanism of obstacle in G. elata continuous cropping had not been solved. The planting of G. elata-Phallus impudicus is a new sequential planting pattern adopted in Guizhou province, but the effect of the cultivation on soil microbial community structure is still unclear. In this study, we collected four soil samples for the research including the soil without planted G. elata as control(CK), rhizosphere soil samples tightly adhering to the G. elata surface(GE), rhizosphere soil samples tightly adhering to Armillaria which was symbiotic with G. elata(AGE), the rhizosphere soil of P. impudicus planting after G. elata cultivation(PI). In order to explore the mechanism, the research study on the soil of G. elata-P. impudicus by using ITS and 16 S rDNA high-throughput sequencing technologies to detect soil microbial community structure including fungi and bacteria in the soil of CK, AGE, GE and PI. OTU clustering and PCA analysis of soil samples showed that the soil microbial diversity was relatively similar in AGE and GE. And the soil microbial in PI and CK clustered together. The results showed that AGE and GE had similar soil microbial diversity, as well as PI and CK. Compared with CK, the soil microbial diversity and abundance in AGE and GE were significantly increased. But the microbial diversity and abundance decreased in PI compared with AGE and GE. The annotation indicated that the abundance of Basidiomycota, Acidobacteria and Chloroflexi decreased, and that of Ascomycota, Zygomycota and Proteobacteria increased in AGE and GE compared with CK. In contrast to AGE and GE, PI was the opposite. The abundance of Basidiomycota, Acidobacteria and Chloroflexi increased in PI compared with AGE and GE. The abundance of microorganisms in the soil of PI and CK was similar. In addition, the co-culture of Armillaria and P. impudicus indicated that P. impudicus had obvious antagonistic effects on the growth of Armillaria. Therefore, it is speculated that the mechanism of G. elata-P. impudicus planting pattern related to the change of soil microbial. And we supposed that P. impudicus might inhibit the growth of Armillaria and change the soil microbial community structure and the abundance of soil microbial. And the soil microbial community structure was restored to a state close to that of uncultivated G. elata. Thus, the structure of soil microbial community planting G. elata could be restored by P. impudicus planting.

[Dissection of seed development of Gastrodia elata at different temperatures].

The study is aimed to create seed materials and dissect the molecular mechanism of sexual propagation of Gastrodia elata. In this research, thirteen characteristics of flowers, flower stem, fruits, seeds and embryo of G.elata f. glauca and G.elata f. elata after bolting at room temperature(RT) and constant temperature(CT, 22 ℃) were determined. It was found that the constant temperature condition could prolong the bolting duration of G.elata and increased the number of flowers, while the variety of G.elata only affected the bolting duration, but had no effect on the number of flowers, and the G.elata f. elata was more likely to bolting than the G.elata f. glauca. The variety of G.elata was the main factor affecting the time of dehiscent fruit of G.elata, the temperature was the main factor affecting the fruits number and fruits diameter, and the constant temperature was more conducive to the fruits shape of G.elata than the room temperature. There was no significant difference in seed phenotype of G.elata varieties, but the seed embryo of G.elata seeds cultivated at constant temperature was fuller than that of G.elata cultivated at room temperature, and temperature had less influence on the seed phenotype of G.elata. But it was interesting to find that temperature and varieties had greater influence on the seed embryo of G.elata, constant temperature cultivation was more conducive to the formation of seed embryo of G.elata, and more the seed embryo of G.elata f. elata was easier to form than the seed embryo of G.elata f. glauca. However, the development of seeds and embryos of G.elata was significantly affected, and the development of seeds and embryos of G.elata f. glauca was more sensitive to temperature than G.elata f. elata. The research suggested that it is advisable for G.elata to produce seed materials by bolting at constant temperature(22 ℃).

Synchronous bilateral clear cell renal cell carcinoma and prostate gland carcinoma on 18F-FDG PET/CT in one patient.

We present a case of synchronous bilateral renal cell carcinoma and prostate carcinoma in a 62 year old man. In a fluorine-18-fluorodeoxyglucose positron emission tomography (18F-FDG PET) scan, the left renal mass showed intense 18F-FDG uptake with maximum standardized uptake volume (SUVmax) of 8.12, while uptake in the right renal mass was minimal with SUVmax of 2.99. Fluorine-18-FDG uptake in the prostate gland lesion was moderate with SUVmax of 4.19. Histopathologically, both renal tumors were clear cell renal cell carcinoma with International Society of Urological Pathology (ISUP) Grade 2 and 3 for the right and left kidney, respectively and prostate gland lesion was typical prostate gland carcinoma.

[Quality assessment of soil fertility of planting Chrysanthemum morifolium areas of Macheng city based on minimum data set].

A minimal data set( MDS) for soil fertility evaluation of Chrysanthemum plantation areas of Macheng city was established by principal component analysis( PCA) combined with Norm values of soil fertility indices and correlation coefficients among indices. A radar map was used to visually reflect the fertility level of individual indicators. Then,the comprehensive index model was used to calculate the soil fertility quality index( SFQI),and the values of SFQI was used to cluster,and the results showed that MDS was composed of five indicators: organic matter( OM),total phosphate( TP),available phosphorus( Av P),available magnesium( Av Mg) and available ferrum( Av Fe). Radar maps showed that the fertility of available phosphorus( Av P) and available copper( Av Cu) was mostly different in the two town,and the fertility of available ferrum( Av Fe) is smallest different. Except for the effective manganese( Av Mn) fertility level of Huangtugang town was higher than that of Futianhe town,the rest were lower than that of Futianhe town. Through analysis,the sensitivity of SFQI value calculated by taking the contribution rate of MDS index in the principal component of the whole data set( TDS) as the weight was the highest,MDS could better replace TDS. The value of SFQI-MDS ranged from 0. 353 to 0. 833,with an average value of 0. 604 and a coefficient of variation of 22%. The results of SFQI-MDS clustering showed that soil fertility could be divided into four categories: grade Ⅰ( 0. 727-0. 833) was superior,accounting for 25. 0%,grade Ⅱ( 0. 615-0. 681)was good,accounting for 29. 2%,mainly distributed in Futianhe Town,grade Ⅲ( 0. 494-0. 589) was medium,accounting for29. 1%,and grade Ⅳ( 0. 353-0. 419) was poor,accounting for 16. 7%,mainly distributed in Huangtugang town. Soil fertility of Futianhe town was better than that of Huangtugang town. It is suggested that boron fertilizer and potassium fertilizer should be supplemented to Chrysanthemum morifolium in production practice,and the amount of phosphate fertilizer,magnesium fertilizer and nitrogen fertilizer should be increased appropriately. At the same time,the amount of organic fertilizer should be increased to enhance soil fertility and improve soil physical and chemical properties.

[Comparison of agronomic traits of Panax notoginseng between traditional cultivated fields and new cultivated fields].

OBJECTIVE: In order to provide scientific basis for introduction breeding and production regionalization of Panax notoginseng, the environment of producing area, agronomic traits and medicinal material output were investigated. METHOD: Using field survey sampling at harvest time, agronomic traits indicators of leaf segment, stem segment and roots segment etc. of commodity P. notoginseng, longitude and latitude, elevation, soil type, landform of producing area were measured and observed. RESULT: The P. notogiseng cultivation was expanding from traditional area like Wenshang to new areas as Honghe, Kunming, Qujing at large scale. Comparing with traditional cultivated fields, the elevation of new fields, which are red soil of moderate or low mountain slopes and gentle hills between 1 800-2 130 m, increases markedly. The agronomic traits of new cultivated fields such as plant height, stem diameter, the ground and underground biomass were better than those of traditional cultivated fields in varying degree. Furthermore, the root weight, taproot weight, taproot length and other agronomic index augmented more than 20%. Comparison among different cultivated fields showed P. notoginseng of Honghe Shiping Niujie, Kunming Shilin Guishan, Honghe Jiangshui Guanting etc. had better agronomic traits, the plant were thick and tall, the taproot and the top of reed were large, the number of root was more. Inversely, P. notoginseng of Kunming Guandu Xiaoshao, Guangxi Jingxi Lutong, Wenshang Yanshan Jiangna were thin and small. Cluster analysis showed that cultivated fields of P. notoginseng which was across clustered by traditional and new cultivated fields can be divided into three groups, including a high-yielding region insist of three new cultivated fields Honghe Shiping Niujie, Kunming Shilin Guishan, Honghe Jiangshui Guanting and one traditional cultivated field Wenshan Yanshan Jiangna. Correlation analysis indicated that the size and weight of taproot and top of reed were significantly positive correlated with plant height, stem diameter, leaf size, leaf weight and stem weight. Regression analysis pointed out that stem diameter, leaf width, leaf length/width, leaf weight were the main factors affecting the dry weight of taproot, and the main factors influencing the dry weight of top of reed were plant height, petiole, leaf length, leaf width and other agronomic traits. CONCLUSION: From agronomic traits, P. notoginseng in new cultivated fields were more robust than that in traditional fields, but besides the length of taproot, the difference of rest agronomic traits didn't reach the significant level. It is suggested that, in the west and the north of traditional area, the red soil land of moderate or low mountain slopes and gentle hills where the elevation is between 1 800-2 130 m is suitable for cultivating P. notoginseng. Honghe Shiping Niujie, Honghe Jianshui Guanting, Kuming Shilin Gongshan can be used as vital development area for commodity P. notoginseng.

[Double mulching application for Panax notoginseng growing seedlings].

OBJECTIVE: In order to improve the irrigation for Panax notginseng growing seedlings, different mulching ways were carried out to investigate the effects of double mulching. METHOD: Field experiment was applied to study soil moisture, soil temperature and bulk density of different mulching ways while the germination rate and seedlings growth also were investigated. RESULT: Compared with the traditional single mulching with pine leaves or straw, double mulching using plastic film combined with pine leaves or straw could reduce 2/3 volumes of irrigation at the early seedling time Double mulching treatments didn't need to irrigate for 40 days from seeding to germination, and kept soil moisture and temperature steady at whole seedling time about 30% and 9.0-16.6 degrees C, respectively. The steady soil moisture and temperature benefited to resist late spring cold and germinate earlier while kept germination regularly, higher rate and seedlings quality. In contrast, single mulching using pine leaves or straw had poor soil moisture and temperature preserving, needed to irrigate every 12-day, meanwhile dropped the germination and booming time 14 days and 24-26 days, respectively, reduced germination rate about 11.3%-8.7%. However, single pine leaves mulching was better than straw mulching. In addition, though better effects of soil moisture and temperature preserving as well as earlier and higher rate of germination with single plastic films mulching had, some disadvantages had also been observed, such as daily soil temperature changed greatly, seedling bed soil hardened easily, more moss and weeds resulted difficulty in later management. CONCLUSION: To the purpose of saving water and labor as well as getting higher germination rate and seedlings quality, double mulching using plastic films combined pine leaves at the early time and single mulching removing plastic films at the later time is suggested to apply in the growing seedlings of P. notoginseng.

[Different proportion of potassium chloride and potassium sulphate application on cultivation of Panax notoginseng].

In order to make sure whether Panax notoginseng is sensitive to chloridion and guide fertilization in planting of P. notoginseng, the effects of the different proportion of potassium chloride (KCl) and potassium sulfate (K2SO4) on the yield, quality of P. notoginseng were studied. The results showed that K fertilizer significantly improved the growth of P. notoginseng and increased the biomass per plant or per pot and the content of N, P, K and the content of saponin. In cases of conditions such as potassium, and the effects of K2SO4 on increasing the petiole length, leaf size, rhizome length, root length, and content and accumulation of Ginsenoside Rg1 were better than those of KCl. While compared with K2SO4, KCl was more conducive to augmenting height, root width, the biomass of shoot, rhizome, root and the content of Ginsenoside Rb1 and Rd. There was not remarkable difference in agronomic characters, biomass and the content of N, P, K among KCl, K2SO4 and the combination of KCl and K2SO4. However, the content of saponin of the treatment with combination of KCl and K2SO4 was significant higher than that of single KCl or K2SO4 treatments. K fertilizer significantly increased yield and the content of saponins. And P. notoginseng was not sensitive to chloridion. KCl increased the yield and the content of saponins of P. notoginseng as well as K2SO4, and the combination treatment was superior to single treatment. It is recommended that the KCl should be adopted in production, to reduce the cost of potash fertilizer.

[The influence of heartbeat acceptance window settings on left ventricular function and perfusion parameters].

OBJECTIVE: To evaluate the influence of heartbeat acceptance window settings on left ventricular function and perfusion parameters of the arrhythmia patients during gated myocardial perfusion imaging. METHODS: Twenty eight fibrillation patients were consecutively recruited to undergo myocardial perfusion SPECT. The Concurrent Imaging software was used to create 3 separate SPECT studies with heartbeat acceptance window of 20%, 60% and 100% respectively. The software created the 3 studies separately rather than a rearrangement of an original list-mode acquisition. After reconstruction by Astonish, end-diastole volume (EDV) and end-systolic volume (ESV), left ventricular ejection fraction (LVEF), sum stress score (SSS), and sum rest score (SRS) were calculated with Quantitative Gated SPECT (QGS). Analyses of variance were performed using SPSS to compare the differences in EDV, ESV, EF, SSS, and SRS among the three studies. RESULTS: 85.7% of the 28 patients had abnormal perfusing. No statistical differences were found in EDV, ESV, EF, SSS, and SRS among the 3 studies. But the collection time was 40.5 min, 25.6 min and 15.0 min for heartbeat acceptance window of 20%, 60% and 100% respectively. CONCLUSION: Heartbeat acceptance window setting does not have a significant effect on EDV, ESV, EF, SSS, and SRS values. The wider the window is set, the shorter the collection time is.

[Construction and expression of replication-deficient recombinant adenovirus vector with hNET gene by adeasy-1 system].

OBJECTIVE: To construct and identify the recombinant replication deficient adenovirus vector which codes for human Norepinephrine Transporter (hNET) gene by using the method of homogenous recombination in bacteria. METHODS: hNET gene was obtained from the recombinant plasmid pCMV5 via Kpn I + Xba I digestion, and subcloned into E1 deleted expression plasmid pAdtrack-CMV shuttle vector, forming transfer vector pAdtrack-CMV-hNET. Then it was linearized with Pme I followed by homologous recombination with bone plasmid pAdEasy-1 in BJ5183 cells to generate recombinant plasmid Ad-hNET. The DNA of identified Ad-hNET was digested with Pac I and transfected to HEK293 cells by liposome-mediated method to package recombinant adenovirus. The PCR technique was applied to detect the target gene and Western Blotting to verify the expression of hNET. The titre of the Ad-hNET was measured with the aid of green fluorescence protein (GFP) expression after multiplication and purification. RESULTS: By sequencing, it was confirmed that the product was the gene of hNET. PCR test, restriction endonuclease digestion and Western Blotting confirmed the successful construction of the recombinants Ad-hNET. The titre of purified recombinant adenovirus Ad-hNET was 1.2 X 10(10) pfu/mL. CONCLUSION: The recombinant adenovirus with the hNET gene was constructed successfully. It will be helpful for the further investigation of its potentiality to be applied in the tumors targeted therapeutic strategy.

[Preparation and biodistribution of VIP-125I-ASON].

OBJECTIVE: To prepare VIP-125I-ASON and investigate the possibility of using it as an agent for diagnostic imaging and therapy of colon carcinoma. METHODS: The iodination of a 15-base single-stranded antisense oligonucleotide (ASON) complementary to C-myc oncogene mRNA was carried out in the presence of TICl3. The radiolabeled oligonucleotide was complexed with a VIP-polylysine conjugate under certain condition. 3-5 microCi VIP-125I-ASON was injected into the tail vein of the BALB/c nude mice bearing transplanted HT29 colon carcinoma; the nude mice were killed at specific intervals after injection, and the biodistrbution of VIP-125I-ASON in the organs were calculated. RESULTS: The biodistributed experiment showed that the 125I-ASON was excreted by kidney mostly and by liver and spleen in part. The results of studies after the injection of VIP-125I-ASON differed from those of unconjugated 125I-ASON. The conjugation of VIP to the ASON resulted in a decrease in the plasma clearance of the radiopharmaceutical, which may be due to the reduction in the renal clearance of the ASON. The highest uptake of tumor tissue (5.89% ID/g at 2 h) was significantly higher than that in nude mice given unconjugated ASON (P < 0.05). Tumor to blood ratios and tumor to muscle ratios were optimal at 4 h. CONCLUSION: VIP-125I-ASON has desirable stability and higher uptake in tumor. It may provide a new sensitive mean for diagnostic antisense imaging and radiotherapy of tumor in the future.

Receptor binding characteristics and cytotoxicity of insulin-methotrexate.

AIM: To characterize the receptor binding affinity and cytotoxicity of insulin-methotrexate (MTX) for the potential utilization of insulin as carriers for carcinoma target drugs. METHODS: MTX was covalently linked to insulin. Insulin-MTX conjugate was purified by Sephadex G-25 column and analyzed by high performance liquid chromatography. Hepatocellular carcinoma cell membrane fractions were isolated by sucrose density gradient centrifugation. Competitive displacement of (125)I-insulin with insulin and insulin-MTX binding to insulin receptors were carried out. Cytoreductive effect of insulin-MTX on human hepatoma BEL7402 cells and human hepatocyte cell line HL7702 was evaluated using the MTT assay. RESULTS: Insulin-MTX competed as effectively as insulin with (125)I-insulin for insulin receptors. The values of Kd for insulin-MTX and insulin were 93.82+/-19.32 nmol/L and 5.01+/-1.24 nmol/L, respectively. The value of Kd for insulin-MTX was significantly increased in comparison with insulin (t=7.2532, n=4, P<0.005). Insulin-MTX inhibited the growth of human hepatoma cells (BEL7402) almost as potently as MTX. The inhibitory effect reached a peak on the 5 th day when the growth of cells was inhibited by 79% at a concentration of 5.0 microg/mL insulin-MTX. Treatment with 5.0 microg/mL of MTX and 5.0 microg/mL of insulin-MTX merely resulted in inhibition of HL7702 cells by 31.5% and 7.8% on the 5 th day. CONCLUSION: Insulin-MTX specifically recognizes insulin receptors and inhibits the growth of BEL7402 cells. These results suggest that insulin can be used as a carrier in receptor mediated carcinoma-targeting therapy.

Study on the possibility of insulin as a carrier of IUdR for hepatocellular carcinoma-targeted therapy.

AIM: To evaluate the possibility of using insulin as a carrier for carcinoma-targeted therapy mediated by receptor, and to investigate the expression of insulin receptor in human hepatocellular carcinoma and the receptor binding characteristics of insulin-IUdR (iododeoxyuridine). METHODS: IUdR was covalently conjugated to insulin. Receptor binding assays of (125)I-insulin to human hepatocellular carcinoma and its adjacent tissue were performed. Competitive displacements of (125)I-insulin by insulin and insulin-IUdR to bind to insulin receptor were respectively carried out. Statistical comparisons between the means were made with paired t-test at a confidence level of 95 %. RESULTS: The data indicated that there were high- and low- affinity binding sites for (125)I-insulin on both hepatocellular carcinoma and its adjacent tissue. Hepatocellular carcinoma had a significantly higher Bmax for high affinity binding site than its adjacent liver tissue (P<0.05, t=2.275). Insulin-IUdR competed as effectively as insulin with (125)I-insulin for binding to insulin receptor. Values of IC(50)1, C(50)2, KI1 and KI2 for insulin-IUdR were 11.50+/-2.83 nmol x L(-1), 19.35+/-5.11 nmol x L(-1), 11.26+/-2.65 nmol x L(-1) and 19.30+/-5.02 nmol x L(-1) respectively, and for insulin were 5.01+/-1.24 nmol x L(-1),17.75+/-4.86 nmol x L(-1), 4.85+/-1.12 nmol x L(-1) and 17.69+/-4.81 nmol x L(-1), respectively. Values of IC(50)1 and KI1 for insulin-IUdR were significantly higher than that for insulin (P<0.01, t=4.537 and 4.813). CONCLUSION: It is possible to use insulin as a carrier for carcinoma-targeted therapy mediated by receptor.