A newly identified cluster of glutathione S-transferase genes provides Verticillium wilt resistance in cotton.

As the largest cultivated fiber crop in the world, cotton (Gossypium hirsutum) is often exposed to various biotic stresses during its growth periods. Verticillium wilt caused by Verticillium dahliae is a severe disease in cotton, and the molecular mechanism of cotton resistance for Verticillium wilt needs to be further investigated. Here, we revealed that the cotton genome contains nine types of GST genes. An evolutionary analysis showed that a newly identified cluster (including Gh_A09G1508, Gh_A09G1509 and Gh_A09G1510) located on chromosome 09 of the A-subgenome was under positive selection pressure during the formation of an allotetraploid. Transcriptome analysis showed that this cluster participates in Verticillium wilt resistance. Because the Gh_A09G1509 gene showed the greatest differential expression in the resistant cultivar under V. dahliae stress, we overexpressed this gene in tobacco and found that its overexpression resulted in enhanced Verticillium wilt resistance. Suppression of the gene cluster via virus-induced gene silencing made cotton plants of the resistant cultivar Nongda601 significantly susceptible. These results demonstrated that the GST cluster played an important role in Verticillium wilt resistance. Further investigation showed that the encoded enzymes of the cluster were essential for the delicate equilibrium between the production and scavenging of H2 O2 during V. dahliae stress.

[Spray drying process of extract of Wenjing Zhitong Prescription based on mixture design experiment].

To improve the spray drying effect of extract of Wenjing Zhitong Prescription, this study takes the yield, hygroscopic property and the fluidity of dry powder as indexes to screen out auxiliary materials, and the proportion of the auxiliary materials was optimized based on the mixing design experiment; based on that, HPLC method was established for the determination of glycyrrhizin and 6-gingerol in spray powder, the yield of spray powder and the retention rate of the two index components were taken as indexes to further optimize the spray drying parameters. The finally selected auxiliary materials were light magnesium oxide, maltodextrin and silica, and regression equations of dry powder yield, moisture absorption rate, angle of rest with proportion of auxiliary materials were established, and the optimized proportion of auxiliary materials was dry paste-light magnesium oxide-maltodextrin-silica=0.5∶0.305∶0.145∶0.05; according to the optimized drying process parameters of Wenjing Zhitong Prescription, initial temperature was 60 ℃, air inlet temperature was 130 ℃, air flow rate was 35 m~3·h~(-1), atomizing pressure was 40 mm, and liquid inlet speed was 4.5 mL·min~(-1). Under these conditions, the dry powder yield was 90.28%, the retention rate of glycyrrhizin was 74.51%, and the retention rate of 6-gingerol was 72.10%. In this study, optimized auxiliary materials can improve the yield of spray drying and the property of spray powder, and the optimized processing conditions were good for retaining the unstable gingerol components, which can lay a foundation for the further preparation research of meridian warming and pain relieving prescriptions, and provide reference for extract of other traditional Chinese medicine extracts that are difficult to spray drying.

[Effect of andrographolide on quorum sensing and relevant virulence genes of Candida albicans].

OBJECTIVE: To investigate the effect of andrographolide (AG) on quroum sensing (QS) and relevant virulence genes of Candida albicans. METHOD: Gas-chromatography-mass spectrometry (GC-MS) was applied to detect the changes in the content of farnesol and tyrosol in C. albicans intervened by AG. The real-time quantitative PCR (qRT-PCR) was adopted to inspect the expressions of relevant virulence genes such as CHK1, PBS2 and HOG1 regulated by QS. RESULT: At 2 h after the growth of C. albican, the farnesol and tyrosol secretions reduced, without notable change after intervention with AG. The secretions were highest at 12 h and decreased at 24 h. After the intervention with different concentrations of AG, the farnesol content reduces, whereas tyrosol increased, indicating a dose-dependence, particularly with 1 000 mg x L(-1) AG. qRT-PCR revealed that 1 000 mg x L(-1) AG could down-regulate CHK1 by 2.375, 3.330 and 4.043 times and PBS2 by 2.010, 4.210 and 4.760 times, with no significant change in HOG1. CONCLUSION: AG could inhibit the farnesol secretion, promote the tyrosol secretion and down-regulate QS-related virulence genes CHK1 and PBS2 expressions.

[Anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction on Candida glabrata].

OBJECTIVE: To investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata. METHOD: Serial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7. RESULT: The MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment. CONCLUSION: EAHD could effectively inhibit adherence of C. glabrata.

[Effect of Huanglian Jiedu decoction in combination with fluconazole on ergosterol of fluconazole-resistant Candida albicans].

OBJECTIVE: To investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans. METHOD: The minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively. RESULT: MICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group. CONCLUSION: The combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

[Inhibitory effects of ethyl acetate extract of Huanglian Jiedu decoction on hyphae development of Candida albicans].

OBJECTIVE: To investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on hyphae development of Candida albicans. METHOD: Inverted microscope, fluorescence microscope, SEM were applied to inspect the Morphological change of C. albicans treated by EAHD at different concentrations. Solid agar plate was utilized to evaluate the colony morphology. Quantitative Real-ime PCR(qRT-PCR) was adopted to observe the expression of hyphae-specific genes such as HWP1, ALS3, UME6, CSH1, SUN41, CaPDE2. RESULT: EAHD with concentration of 312 and 1 250 mg . L-1 could inhibit formation of hyphae and colony morphology. The expression of HWP1, ALS3, UME6, CSH1 were downregulated 4. 13, 3. 64, 2. 46, 2. 75 folds ,while the expression of SUN41 were upregulated 7. 26 folds, CaPDE2 keep unchanged. CONCLUSION: EAHD could inhibit formation of hyphae and colony morphologies of C. albicans through downregulating HWP1, ALS3, UME6 and CSH1.

[Study on inhibitory effect of different extract fractions from longdan xiegan decoction on biofilms of Candida albicans].

OBJECTIVE: To observe the inhibitory effect of different extract fractions from Longdan Xiegan decoction on biofilms of Candida albicans, and discuss its possible mechanism. METHOD: The micro-dilution method and the XTT reduction assay were adopted to explore the antifungal activity of different extract fractions from Longdan Xiegan decoction and detect the inhibitory effect of different extracts on biofilms of C. albicans. The expression quantity of the adhesion related gene ALS1 and hypha formation SUN41 were detected by qRT-PCR. RESULT: The MICs of extracts from Longdan Xiegan decoction, including petroleum ether, water, butanol, methanol and ethyl acetate, against C. albicans were > 1 000, > 1 000, > 1 000, 125, 125 mg x L(-1). The SMIC50 against biofilms of C. albicanswere > 1 000, > 1000, > 1 000, 500, 500 mg x L(-1). The SMIC50 were > 1 000, > 1 000, > 1 000, > 1 000 and 1 000 mg x L(-1). 1 000 mg x L(-1) ethyl acetate extracts could considerably inhibit the expression of the adhesion related gene ALS1 and hypha formation SUN41. CONCLUSION: The ethyl acetate extract showed the greatest activity against Candida albicans biofilms.

[Effect of andrographolide on Candida albicans biofilm dispersion].

Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.

[Effect of andrographolide derivative Yanhuning on in vivo Candida albicans biofilms in rats].

OBJECTIVE: To investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats. METHOD: The rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65. RESULT: The YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65. CONCLUSION: YHN could inhibit C. albicans biofilms in rats.

[Choroidal thickness after half-dose verteporfin photodynamic therapy in chronic central serous chorioretinopathy].

OBJECTIVE: To observe the choroidal thickness in eyes with chronic central serous chorioretinopathy(CSC) after half-dose photodynamic therapy(PDT). METHODS: Case series study. Consecutive 30 patients (35 eyes) with chronic CSC admitted from October 2010 to October 2011 in Zhongshan Ophthalmic Center were recruited in this study. Optical coherence tomography(OCT),fundus fluorescein angiography, indocyanine green angiography (ICGA) were performed in each patient to confirm the diagnosis. ICGA-guided PDT was performed in each patient. Choroidal thickness at subfoveal and 1500 µm superior, inferior, nasal and temporal to the fovea were measured before PDT, 1, 3 and 6 months after PDT using enhanced depth imaging OCT. Data were analyzed with one-way repeated measures ANOVA and LSD-t test when pairwise comparison was needed. RESULTS: Thirty patients, aged from 33 to 66 years old, including 26 males and 4 females (gender ratio 6.5: 1) were recruited. Subfoveal choroidal thickness (SFCT) in chronic CSC eyes was (435.54 ± 102.91) µm at baseline,which decreased significantly to (373.63 ± 106.01) µm, (360.69 ± 111.06) µm and (369.00 ± 108.12) µm at 1 month, 3 months and 6 months after PDT respectively (F = 108.34, P < 0.01). At 6 months after PDT, SFCT was slightly thicker than that at the 3(rd) month,but no statistical significance was noted (LSD-t test: P > 0.05). The changing trends of choroidal thickness at the other 4 measuring points were similar to that of the SFCT. CONCLUSION: Choroidal thickness is reduced within 6 months post PDT in chronic CSC eyes.

[Advance in studies on anti-infection of andrographolide and its derivatives in past 10 years].

Andrographolide (AG) is a main effective ingredient in leaves of Andrographis paniculata. AG and its derivatives have such effects in anti-infection, anti-tumor and immuno-regulation. Particularly, its antibacterial, antiviral and anti-protozoal activities play an increasingly important role in resisting opportunistic infections. On the basis of our studies on AG over the years, we made a summary for the basis and clinical studies on AG and its derivatives over the past 10 years.

Geographical variation of ecological stoichiometry and nutrient resorption in leaves of Pinus koraiensis and Fraxinus mandshurica.

In this study, we sampled leaves of coniferous species Pinus koraiensis and broad-leaved tree species Fraxinus mandshurica from four latitudes in northeastern China to investigate the carbon (C), nitrogen (N), phosphorus (P) stoichiometric characteristics and nutrient resorption efficiency and their potential relationships, as well as their responses to climatic and edaphic factors. The results showed that stoichiometric characteristics were species-specific, and that the C and N contents in leaves of F. mandshurica significantly increased with increasing latitude. The C:N of F. mandshurica and N:P of P. koraiensis were negatively correlated with latitude, but an inverse relationship was found for N:P of F. mandshurica. P resorption efficiency was significantly correlated with latitude in P. koraiensis. The spatial variation of ecological stoichiometry of these two species was mainly affected by climatic factors such as mean annual temperature and precipitation, while that of nutrient resorption was influenced by several soil factors such as soil pH and nitrogen content. Principal component analysis showed that P resorption efficiency of P. koraiensis and F. mandshurica was significantly negatively correlated with N:P, but positively correlated with P content. N resorption efficiency showed significantly positive correlation with P content but negative correlation with N:P in P. koraiensis. Compared with P. koraiensis, F. mandshurica was more inclined to fast investment and return in terms of leaf traits.

Reirradiation with stereotactic body radiotherapy for primary or secondary lung malignancies: Tumor control probability and safety analyses.

BACKGROUND: Reirradiation with stereotactic body radiotherapy (SBRT) for patients with primary or secondary lung malignancies represents an appealing definitive approach, but its feasibility and safety are not well defined. The purpose of this study was to investigate the tumor control probability (TCP) and toxicity for patients receiving reirradiation with SBRT. PATIENTS AND METHODS: Eligible patients with recurrence of primary or secondary lung malignancies from our hospital were subjected to reirradiation with SBRT, and PubMed- and Embase-indexed articles were reviewed. The patient characteristics, pertinent SBRT dosimetric details, local tumor control, and toxicities were extracted. The logistic dose-response models were compared for TCP and overall survival (OS) in terms of the physical dose and three-, four-, and five-fraction equivalent doses. RESULTS: The data of 17 patients from our hospital and 195 patients extracted from 12 articles were summarized. Reirradiation with SBRT yielded 2-year estimates of 80% TCP for doses of 50.10 Gy, 55.85 Gy, and 60.54 Gy in three, four, and five fractions, respectively. The estimated TCP with common fractionation schemes were 50%, 60%, and 70% for 42.04 Gy, 47.44 Gy, and 53.32 Gy in five fractions, respectively. Similarly, the 2-year estimated OS was 50%, 60%, and 70% for 41.62 Gy, 46.88 Gy, and 52.55 Gy in five fractions, respectively. Central tumor localization may be associated with severe toxicity. CONCLUSIONS: Reirradiation with SBRT doses of 50-60 Gy in 3-5 fractions is feasible for appropriately selected patients with recurrence of peripheral primary or secondary lung malignancies, but should be carefully considered for centrally-located tumors due to potentially severe toxicity. Further studies are warranted for optimal dose/fractionation schedules and more accurate selection of patients suitable for reirradiation with SBRT.

Regioselective synthesis of fraxinellone-based hydrazone derivatives as insecticidal agents.

In continuation of our program aimed at the discovery and development of natural products-based insecticidal agents, twenty-three new fraxinellone-based hydrazone derivatives were smoothly prepared from fraxinellone via regioselectively allylic oxidation in the presence of selenium dioxide or chromium trioxide under microwave irradiation and subsequent condensation with hydrazides or hydrazines. Their insecticidal activity was evaluated against the pre-third-instar larvae of Mythimna separata Walker in vivo. Especially compounds 6d and 7a displayed the most pronounced insecticidal activity compared with toosendanin, a commercial botanical insecticide derived from Melia azedarach.

[Toxical research of Kudiezi injection in safety re-evaluation].

OBJECTIVE: To explore the toxic effect on mouse administrated Kudiezi injection multy times a day, and on rats repeat administrated for many days. METHOD: Mouse tail intravenous injection of Kudiezi, 0.04 mL x g(-1), 3 times a day, rats tail intravenous injection of Kudiezi, 20, 10, 4 mL x kg(-1), once a day, for 6 weeks. RESULT: There is no abnormal to the mouses administrated many times a day. The rats administrated large doses of drug for many days have certain effects on hematology, blood biochemistry. Some animals appear liver, kidney lesions mild, injection local appear haemorrhage, edema and inflammatory reaction. CONCLUSION: The mouse which was intravenous injection in the dose of 180 times Kudiezi injection as much as people used, revealed no toxicity reaction. Repeated large-dose administration, rats caused by lesions of the main target organs may be for kidney, liver. But the recovery result on liver, kidney toxicity was reversible, no delayed toxicity. At the same time, large doses of long-term administration of local have a certain irritation. Tips the medication should be under the guidance of doctors, and pay attention to replace the injection site. This research will provide safety basis for the clinical use of Kudiezi injection.

HN1L/AP-2γ/PLK1 signaling drives tumor progression and chemotherapy resistance in esophageal squamous cell carcinoma.

Hematological and neurological expressed 1 like (HN1L) is a newly identified oncogene in lung cancer and hepatocellular carcinoma recently identified by our team, but its roles in the development and treatment of esophageal squamous cell carcinoma (ESCC) remain incompletely cataloged. Here, using ESCC tissue array and public database analysis, we demonstrated that HN1L was highly expressed in ESCC tissues, which was associated with tumor tissue invasion, poor clinical stage and short survival for ESCC patients. Loss- and gain-of-function studies in ESCC cells revealed that HN1L enhances ESCC cell metastasis and proliferation in vitro and in mice models. Moreover, high level of HN1L reduces the sensibility of ESCC cells to chemotherapeutic drugs, such as Docetaxel. Mechanism studies revealed that HN1L activated the transcription of polo-like kinase 1 (PLK1) by interacting with transcription factor AP-2γ, which increased the expression of malignancy related proteins Cyclin D1 and Slug in ESCC cells. Blocking PLK1 with inhibitor BI-2356 abrogated the oncogenic function of HN1L and significantly suppressed ESCC progression by combining with chemotherapy. Therefore, this study demonstrates the vital pro-tumor role of HN1L/AP-2γ/PLK1 signaling axis in ESCC, offering a potential therapeutic strategy for ESCC patients with high HN1L by blocking PLK1.

[Corrigendum) RNA interference­mediated FANCF silencing sensitizes OVCAR3 ovarian cancer cells to adriamycin through increased adriamycin­induced apoptosis dependent on JNK activation.

After the publication of the article, an interested reader drew to the authors' attention that there appeared to be a pair of overlapping data panels in Fig. 4C on p. 1726 [specifically, the 'Untransfected' and 'Control shRNA' data panels for the ADM (24 h) experiments]. The authors have consulted their original data, and have realized that this figure was inadvertently assembled incorrectly. Furthermore, they have noticed that Fig. 1 on p. 1724 also contained errors that arose during its assembly; essentially, several of the data panels in Fig. 1C, showing the detection of FANCD2 focus formation via immunofluorescence experiments, were selected inappropriately. The corrected versions of Figs. 1 and 4, containing the corrected data panels for Figs. 1C and 4C respectively, are shown on the next page. Note that these errors did not affect the results or the conclusions reported in this work. The authors all agree to this Corrigendum, and are grateful to the Editor of Oncology Reports for allowing them to have the opportunity to correct these mistakes. Lastly, the authors apologize to the readership for any inconvenience these errors may have caused. [Oncology Reports 29: 1721­1729, 2013; DOI: 10.3892/or.2013.2295].

Elucidation of the structural changes of sesame hull hemicelluloses during roasting.

In this investigation, the hemicellulose fractions were isolated from sesame hulls before and after roasting at 160 °C, 180 °C, 200 °C, and 220 °C. The structural properties of the hemicelluloses were comparatively investigated by analytical techniques including FT-IR, sugar compositions analysis, molecular weight analysis, thermal analysis, and NMR. Roasting at temperatures from 180 to 220 °C resulted in higher yields of hemicelluloses subsequently isolated from sesame hulls by the alkaline method. The results of sugar analysis and average molecular weight analysis of the hull hemicelluloses revealed that the side chain sugar units such as arabinose and rhamnose of the hemicelluloses are more easily degraded after roasting during the roasting. That the main chains of hemicelluloses in the cell wall of sesame seed hulls did not change significantly. The investigation provides new information about the mechanisms of the structural changes that occur in sesame hull hemicelluloses during roasting.

Effects of various oil extraction methods on the gelatinization and retrogradation properties of starches isolated from tigernut (Cyperus esculentus) tuber meals.

Gelatinization and retrogradation characteristics of starches from tigernut (Cyperus esculentus) tuber before and after various oil extraction processes were studied in this investigation. The results indicated that starches isolated from tigernut tuber after the various oil extraction processes varied significantly in gelatinization and retrogradation properties. The starches isolated from the cakes of tigernut tuber after hot press extraction exhibited higher retrogradation tendency and relatively less shear-thinning than other starch samples. The results of FT-IR, XRD, and NMR analysis indicated that oil extraction had an unfavorable influence on starch retrogradation, which may be due to structural changes caused by oil extraction processes. In particular, oil extraction led to more efficient packing of double helices in the crystalline lamella of the starches during storage. Retrogradation of the starch gels also reduced the water holding capacities of the starches. The starch sample isolated from the cake after cold press extraction exhibited the highest water absorption capacity among the five samples for all storage times. This investigation provides valuable novel information for the industrial utilization of tigernut tuber starches isolated from meals and cakes after oil extraction.

Comparative study on the diagnostic values of different ultrasound technologies for malignant thyroid nodules.

The present study compared and analyzed the diagnostic values of B-mode ultrasound and elasticity imaging technology for the identification of benign and malignant thyroid nodules. Ninety-four patients who were diagnosed with thyroid nodules by ultrasound were chosen. All patients were checked with B-mode ultrasound and ultrasound elasticity imaging technology before surgeries. Further, the post-operative outcomes were compared with operation pathology. The pathological examination results were taken as the gold standard. The accuracy, sensitivity, specificity, positive predictive value and negative predictive values of combined ultrasound diagnosis were all obviously higher than those by the B-mode ultrasound and the ultrasound elastography method (P<0.05). In conclusion, as a new technology, ultrasound elasticity imaging technology has relatively high diagnositic value in identifying benign and malignant thyroid nodules. With its own advantages and limitations, the ultrasound elasticity imaging technology could effectively complement B-mode ultrasound, and improve the sensitivity, specificity and accuracy of ultrasound diagnosis for thyroid nodules.