[Spatial and temporal characteristics of flue-cured tobacco water requirement and irrigation requirement index in Yunnan Province, China].

Based on the daily meteorological data of 124 agricultural meteorological sites during 1977-2010 in Yunnan Province, using recommended Penman-Monteith formula by FAO, water requirement and irrigation requirement index in the growth period of flue-cured tobacco were calculated to analyze their spatial and temporal characteristics and change patterns. The results showed that water requirements of flue-cured tobacco in root extending, vigorous, mature periods and field growth period during 1977-2010 were 76.73-174.73, 247.50-386.64, 180.28-258.14 and 528.18-764.08 mm, respectively, and the water requirement of vigorous period was the highest. The average irrigation demand index of each period was -0.02, 0.38, 0.17 and 0.26, respectively. Effective precipitation could meet the demand of flue-cured tobacco in root extending period. Water requirement of flue-cured tobacco in Yunnan Province decreased annually, and the rates of water requirement under the climate change trend in the four periods abovementioned were -12. 42, -21.46, -7.17 and -47.15 mm . (10 a)-1, respectively. The smallest irrigation demand index was observed in Dehong, and the largest in Diqing. The irrigation demand indexes of Dehong, Xishuangbanna and Puer regions were negative in flue-cured tobacco field growth period. The reference crop evapotranspiration, water requirement and effective precipitation decreased, but the irrigation requirement and irrigation requirement index increased with the increase of latitude. The effective precipitation decreased, but the irrigation requirement and irrigation requirement index increased with the increase of altitude.

EUS-guided ethanol ablation of insulinomas: case series and literature review.

Surgical resection is a standard treatment for insulinomas; however, it is associated with a high risk of complications and limited to specific suitable candidates. In recent years, endoscopic ultrasound (EUS)-guided ethanol ablation of insulinomas has emerged as a new therapeutic option, especially for elderly patients and candidates unfit for surgery. We aimed to evaluate the feasibility and safety of this technique for insulinomas. Four patients diagnosed with insulinomas based on EUS-fine-needle aspiration and immunohistochemistry results underwent EUS-guided 95% ethanol ablation. A comprehensive literature review was performed to understand the current status of the feasibility, safety, and effects of EUS-guided ethanol ablation of insulinomas. EUS-guided ethanol ablation of insulinomas was successfully completed in all the 4 patients. There were no perioperative or postoperative complications. The patients were discharged at 3 days after the procedure. No recurrence of hypoglycemia or tumors was noted during follow-up (range, 3-6 months). Literature review showed 8 patients with insulinomas who underwent EUS-guided ethanol ablation. All the procedures were successful, with no need for further surgical treatment. Among these reviewed cases, 6 patients had no post-procedural complications, while other 2 patients showed a mild increase in the serum levels of lipase and/or pancreatic enzymes within 48 h post-procedure; furthermore, 1 of these 2 patients presented at a later date with medically controllable hematoma and ulceration. During follow-up, 6 patients remained asymptomatic and normoglycemic, while the 2 patients who presented post-procedural complications developed occasional mild confusion. EUS-guided ethanol ablation of insulinomas is an effective and safe modality, with an acceptable level of post-procedural complications. However, the long-term effects of this new therapeutic option need to be validated in a large randomized controlled trial with longer follow-up.

Establishment of a novel method for primary culture of normal human cervical keratinocytes.

BACKGROUND: Cervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro. METHODS: Normal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity. RESULTS: Our results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%. CONCLUSION: A novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Three novel xanthones from Garcinia paucinervis and their anti-TMV activity.

Phytochemical investigations of the leaves of Garcinia paucinervis resulted in the isolation of three new xanthones 1-3 and five known analogues 4-8. Structural elucidations of 1-3 were performed by spectral methods such as 1D and 2D (HMQC, HMBC, and ROESY) NMR spectroscopy, in addition to high resolution mass spectrometry. Compounds 1-3 showed anti-TMV activities, with inhibition rates above 20%, especially for 1, which had a lower IC50 value of 21.4 µM.

Diversity arrays technology (DArT) for studying the genetic polymorphism of flue-cured tobacco (Nicotiana tabacum).

Diversity arrays technology (DArT) is a microarray-based marker system that achieves high throughput by reducing the complexity of the genome. A DArT chip has recently been developed for tobacco. In this study, we genotyped 267 flue-cured cultivars/landraces, including 121 Chinese accessions over five decades from widespread geographic regions in China, 103 from the Americas, and 43 other foreign cultivars, using the newly developed chip. Three hundred and thirty polymorphic DArT makers were selected and used for a phylogenetic analysis, which suggested that the 267 accessions could be classified into two subgroups, which could each be further divided into 2‒4 sections. Eight elite cultivars, which account for 83% of the area of Chinese tobacco production, were all found in one subgroup. Two high-quality cultivars, HHDJY and Cuibi1, were grouped together in one section, while six other high-yield cultivars were grouped into another section. The 330 DArT marker clones were sequenced and close to 95% of them are within non-repetitive regions. Finally, the implications of this study for Chinese flue-cured tobacco breeding and production programs were discussed.

[Genetic and correlation analysis for agronomic traits in flue-cured tobacco (Nicotiana tabacum L.)].

By using a genetic model including additive and dominance effects and their interaction with environments, 7 agronomic traits were analyzed for a diallel design in 4 environmental conditions with 14 flue-cured tobacco varieties (or breeding lines) and their 41 F1 crosses. It was revealed that additive effects were the major genetic component for plant height, internode length, and width of leaves. Number of leaves and length of leaves were mainly controlled by dominance x environment interaction effects. Additive x environment interaction effects and dominance x environment interaction effects played a major role for girth of stem. Yield performance was mainly controlled by additive effects and dominance x environment interaction effects. The varieties adapted to local environments tended to have highly positive additive effects. Dominance effects and the dominance x environment interaction effects could perform differently in positive or negative direction for many crosses. The breeding program for hybrids should consider the adaptation of hybrids to specific ecological environments. The analysis of correlation among agronomic traits indicated that most of phenotypic, genotypic, additive and dominance correlation coefficients were positive. Additive correlations were predominance in genetic correlations for most pairs of traits. Yield can be improved by indirect selection on plant height.

[Genetic contribution of agronomic traits to yield in flue-cured tobacco (Nicotiana tabacum L.)].

In order to understand the genetic contribution of six agronomic traits to yield, 14 flue-cured tobacco varieties (or breeding lines) and their 41 F1 crosses were used for multivariable conditional analysis. The contribution of additive variance of plant height to yield was larger than other agronomic traits. The largest contribution of dominant variance to yield was due to the length of middle leaves. All agronomic traits investigated had small contribution to yield due to additive x environment interaction effects and dominant x environment interaction effects. No identical trait of different parents showed the largest contribution to additive effect of yield. This could be resulted from the fact that each parent had its own genetic and developmental characterization. The dominant effects of yield were mainly influenced by length of middle leaves in most crosses. Length of middle leaves could be served as ameasurement to indirectly select the cross parent having high dominant effect of yield.