Development and Evaluation of Stable Sugarcane Mosaic Virus Mild Mutants for Cross-Protection Against Infection by Severe Strain.

Sugarcane mosaic virus (SCMV; genus Potyvirus) induces maize dwarf mosaic disease that has caused serious yield losses of maize in China. Cross-protection is one of the efficient strategies to fight against severe virus strains. Although many mild strains have been identified, the spontaneous mutation is one of the challenging problems affecting their application in cross-protection. In this study, we found that the substitution of cysteine (C) at positions 57 or 60 in the zinc finger-like motif of HC-Pro with alanine (A; C57A or C60A) significantly reduced its RNA silencing suppression activity and SCMV virulence. To reduce the risk of mild strains mutating to virulent ones by reverse or complementary mutations, we obtained attenuated SCMV mutants with double-mutations in the zinc finger-like and FRNK motifs of HC-Pro and evaluated their potential application in cross-protection. The results showed that the maize plants infected with FKNK/C60A double-mutant showed symptomless until 95 days post-inoculation and FKNK/C60A cross-protected plants displayed high resistance to severe SCMV strain. This study provides theoretical and material bases for the control of SCMV through cross-protection.

Preparation of camellia oil-based W/O emulsions stabilized by tea polyphenol palmitate: Structuring camellia oil as a potential solid fat replacer.

Water-in-oil (W/O) emulsions with a discontinuous aqueous phase dispersed in a continuous camellia oil phase were prepared by using tea polyphenol palmitate (Tp-palmitate) particle as effective stabilizers and their properties were characterized by droplet size, slip melting point (SMP), stability, microstructure and rheology. The d(4,3) and d(3,2) decreased from 7.96 µm to 4.67 µm and from 5.98 µm to 3.07 µm, respectively, and the SMP rose from 33.73 °C to 38.60 °C when the Tp-palmitate concentration increased from 1.0% to 2.5% (m/v). The storage stability, freeze/thaw stability and thermal stability significantly enhanced and the droplets aggregation progressively increased with the increasing of Tp-palmitate concentration. The liquid camellia oil was transformed into solid-like viscoelastic emulsion gels with a SMP of 38.6 °C when using 2.5% Tp-palmitate as particle stabilizers. This study provides a promising method for production of edible gel-like W/O emulsions using polyphenol-lipid complexes to potentially replace solid fats.

[Efficacy of Huang'e Capsules on benign prostatic hyperplasia with Qi-deficiency, blood stasis and damp heat block: A stratified analysis].

OBJECTIVE: To evaluate the efficacy and safety of Huang'e Capsules in the treatment of BPH with Qi-deficiency, blood stasis and damp heat block. METHODS: This study included 1 456 cases of BPH with Qi-deficiency, blood stasis and damp heat block from 40 hospitals of Zhejiang and Anhui Provinces between June 2014 and July 2017. The patients were aged 40－85 years and treated with Huang'e Capsules at a dose of 4 capsules, tid, for a course of 6 weeks. The IPSSs of the patients were obtained before and after 2, 4 and 6 weeks of medication, and a stratified analysis was made on the factors influencing the therapeutic effects, such as age, the stage of BPH, and concomitant medication of urogenital and reproductive hormone drugs. RESULTS: After 6 weeks of medication, the IPSS of the patients was decreased by 8.4 ± 5.4 (ï¼»42.9 ± 22.7ï¼½ %) as compared with the baseline (19.2 ± 6.8) (P < 0.05), with a total effectiveness rate of 71.9% and an excellence rate of 23.1%. After 6 weeks of medication, the IPSSs of the patients with stage-Ⅰ, -Ⅱ and -Ⅲ BPH were decreased by 7.3 ± 6.8, 6.6 ± 4.3 and 11.0 ± 5.5, with total effective rates of 69.5%, 70.1% and 74.7%, respectively, with statistically significant differences among the three stages (P < 0.05), and so were those in the patients aged ≤50 years (by 7.4 ± 5.8), 51－60 years (by 7.9 ± 5.3), 61－70 years (by 8.7 ± 5.6) and >70 years (by 8.6 ± 5.1), but with no statistically significant differences among different age groups (P > 0.05). At 6 weeks, the IPSS was reduced by 7.1 ± 5.9 by concomitant medication of urogenital and reproductive hormone drugs, 8.7 ± 5.4 by concomitant medication of other drugs, and 8.4 ± 5.3 by non-concomitant medication, with no statistically significant differences among the three types of medication (P > 0.05). CONCLUSIONS: Huang'e Capsules can evidently improve the symptoms of BPH, with the best effect on stage-Ⅲ BPH, and the effect does not differ significantly with age or drug concomitance.

Enzymatic lipophilization of epicatechin with free fatty acids and its effect on antioxidative capacity in crude camellia seed oil.

BACKGROUND: Crude camellia seed oil is rich in free fatty acids, which must be removed to produce an oil of acceptable quality. In the present study, we reduced the free fatty acid content of crude camellia seed oil by lipophilization of epicatechin with these free fatty acids in the presence of Candida antarctica lipase B (Novozym 435), and this may enhance the oxidative stability of the oil at the same time. RESULTS: The acid value of crude camellia seed oil reduced from 3.7 to 2.5 mgKOH g-1 after lipophilization. Gas chomatography-mass spectrometry analysis revealed that epicatechin oleate and epicatechin palmitate were synthesized in the lipophilized oil. The peroxide, p-anisidine, and total oxidation values during heating of the lipophilized oil were much lower than that of the crude oil and commercially available camellia seed oil, suggesting that lipophilized epicatechin derivatives could help enhance the oxidative stability of edible oil. CONCLUSION: The enzymatic process to lipophilize epicatechin with the free fatty acids in crude camellia seed oil described in the present study could decrease the acid value to meet the quality standards for commercial camellia seed oil and, at the same time, obtain a new edible camellia seed oil product with good oxidative stability. © 2016 Society of Chemical Industry.

[Bushen Huoxue Fang promotes the apoptosis of epithelial cells in the prostatic ductal system of rats with benign prostatic hyperplasia].

OBJECTIVE: To investigate the effects of Bushen Huoxue Fang (BSHX) on the apoptosis of epithelial cells in the prostatic ductal system of rats with benign prostatic hyperplasia (BPH) and its possible action mechanism. METHODS: One hundred 3- month-old male Wistar rats were randomly divided into four groups of equal number (control, castrated, BPH model, and BSHX). BPH models were made by subcutaneous injection of testosterone following castration; the rats in the BSHX group were treated intragastrically with BSHX at 2.34 g/ml after modeling, while those in the other two groups with equal volume of saline, all for 37 days. On the 38th day, all the rats were sacrificed and their prostates harvested for detection of the distribution of TGF-beta1 and alpha-actin and the count of positive cells in the prostatic ductal system by immunohistochemical staining. The apoptosis rate of epithelial cells in the prostatic ductal system was determined by TUNEL assay. RESULTS: The expression of TGF-beta1 was significantly increased in the rats of the BSHX group as compared with the BPH models in both the proximal prostatic duct ([15.28 +/- 4.30]% vs [36.42 +/- 8.10]%, P < 0.01) and the distal prostatic duct ([4.42 +/- 2.07]% vs [8.71 +/- 2.28 ]%, P < 0.05), while the expression of alpha-actin in the proximal duct was remarkably higher in the BSHX-treated rats than in the models ([28.14 +/- 7.43]% vs [18.28 +/- 4.07]%, P < 0.01), but lower than in the control animals ([33.57 +/- 6.85]%, P < 0.05). Compared with the control group, the BPH models and BSHX-treated rats both exhibited markedly decreased apoptosis of epithelial cells in the proximal prostatic duct ([39.42 +/- 9.20]% vs [3.86 +/- 1.34]%, P < 0.01, and [31.14 +/- 5.64]%, P < 0.01) and distal prostatic duct ([17.60 +/- 4.86]% vs [3.07 +/- 1.14]%, P < 0.01, and [12.37 +/- 2.25]%, P < 0.05). The apoptosis rate of epithelial cells in the prostatic ductal system was significantly higher in the BSHX-treated rats than in the BPH models (P < 0.01). CONCLUSION: By upregulating the expression of TGF-beta, BSHX can suppress the reduction of smooth muscle cells in the proximal prostatic duct, promote the apoptosis of prostatic epithelial cells, and thus effectively inhibit benign prostatic hyperplasia.

Intraoperative high-dose epinephrine infiltration in cleft palate repair.

OBJECTIVE: Local infiltration of epinephrine before surgical procedures is a well-accepted technique to promote vasoconstriction. Typically, the dose of epinephrine is limited by the co-administration of local anesthetic as well as the risk for arrhythmogenesis and hemodynamic changes. In addition, some controversy exists regarding the acceptable dose of epinephrine given to children. This retrospective review examines the use and safety of "high-dose" epinephrine in palatoplasty at our cleft-craniofacial center. DESIGN: A retrospective review of epinephrine use in primary palatoplasty at a tertiary children's hospital from 2003 to 2007 was performed. Operative and anesthetic records were reviewed for hypertension (systolic blood pressure, >120 or diastolic blood pressure, >70) and tachycardia (>190 beats per min) as defined by the American Heart Association guidelines, as well as dysrhythmias, intraoperative complications, and postoperative complications. RESULTS: A total of 102 patients who underwent consecutive primary palatoplasties performed by a single surgeon were identified. After the induction of anesthesia and before incision, the patients received an initial epinephrine infiltration (without local anesthetic) up to a maximum 10 µg/kg. The average total dose of epinephrine administered during palatoplasty was 12.8 µg/kg (range, 3.2-75.0 µg/kg). Doses up to a maximum of 10 µg/kg were administered as needed at 30-minute intervals. No instances of clinically unstable tachycardia or hypertension occurred. A total of 21.6% of the patients (22/102) experienced an instance of hypertension, and only 13.7% of these (14/102) were related to epinephrine administration. One (1%) postoperative fistula was identified. CONCLUSIONS: Locally infiltrated high-dose epinephrine during palatoplasty can be safely used as a means of vasoconstriction. Doses reaching a maximum of 10 µg/kg, administered as needed at 30-minute intervals, do not seem to be a significant risk for hemodynamic instability, intraoperative complications, or postoperative complications.

Artificial cultivation modes for Dendrobium officinale / 中国中药杂志

Since the beginning of the new century, the artificial cultivation of Dendrobium officinale has made a breakthrough progress. This paper systematically expounds key technologies, main features and cautions of the cultivation modes e.g. bionic-facility cultivation, the original ecological cultivation, and potting cultivation for D. officinale, which can provide useful information for the development and improvement of D. officinale industry.

Early Cambrian ocean anoxia in South China.

The cause of the most marked changes in the evolution of life, which define the first-order stratigraphic boundary between the Precambrian and the Phanerozoic eon, remains enigmatic and a highly topical subject of debate. A global ocean anoxic event, triggered by large-scale hydrogen sulphide (H(2)S) release to surface waters, has been suggested by Wille et al., on the basis of two data sets from South China and Oman, to explain the fundamental biological changes across the Precambrian/Cambrian (PC/C) boundary. Here we report a new precise SHRIMP U-Pb zircon age of 532.3 +/- 0.7 million years (Myr) ago (Fig. 1) for a volcanic ash bed in the critical unit that reflects the ocean anoxic event, the lowermost black shale sequence of the Niutitang Formation in the Guizhou Province, South China. This age is significantly younger than the precise PC/C boundary age of 542.0 +/- 0.3 Myr ago, approximately 10 Myr younger than the extinction of the Ediacaran fauna, and thus challenging the view of a major ocean anoxic event having been responsible for the major changes in the direction of evolution at the PC/C boundary.

Electroacupuncture effects in a rat model of complete Freund's adjuvant-induced inflammatory pain: antinociceptive effects enhanced and tolerance development accelerated.

We have previously shown that electroacupuncture (EA) produced antinociception through the release of endogenous opioid peptides to activate opioid receptors during acute nociception. EA produced tolerance after its prolonged application. It has reported that 100 Hz EA could reduce mechanical hyperalgesia in complete Freund's adjuvant (CFA)-induced inflammatory nociception rats. The present study aims to investigate the antinociceptive effect of EA and the development of EA tolerance in chronic inflammatory nociception rats with CFA injection into the hind paw plantar. The results showed that the antinociceptive effect of 100 Hz EA was significantly enhanced in CFA-induced inflammatory nociception rats. Naloxone at 20 mg/kg could significantly block this antinociceptive effect. Chronic tolerance to EA was developed faster in CFA-induced inflammatory nociception rats than in normal rats. Therefore, 100 Hz EA could enhance antinociceptive effects and accelerate tolerance development in CFA-induced inflammatory nociception rats. The enhancement of EA antinociceptive effect in CFA-induced inflammatory nociception rats might involve the endogenous opioid peptides such as dynorphin.

CCK(B) receptor antagonist L365,260 potentiates the efficacy to and reverses chronic tolerance to electroacupuncture-induced analgesia in mice.

Cholecystokinin octapeptide (CCK-8) is a physiological antagonist of endogenous opioids in the central nervous system (CNS). Our previous work has shown that CCK-8 plays an important role in the development of tolerance to morphine analgesia and electroacupuncture (EA) analgesia in the rat. The present studies were designed to examine whether the CCK(B) receptor is involved in the modulation of EA analgesia and the development of EA tolerance in mice. The latency to flick the tail in the radiant heat was used as index to assess the efficacy of EA analgesia. Subcutaneous (s.c.) injection of the CCK(B) receptor antagonist L365,260 produced a dose-dependent (0.125-2.0 mg/kg) potentiation of the analgesia induced by 100 Hz EA, with a maximal effect occurred at 0.5 mg/kg. In addition, L365,260 (0.5 mg/kg) significantly reversed chronic tolerance to 100 Hz EA in mice. These results suggest that the CCK(B) receptor might play a role in the tonic inhibition of 100 Hz EA-induced analgesia and in the mediation of chronic tolerance to 100 Hz EA in mice. The results opened a way for further investigation of the function of CCK-8 in pain modulation using inbred strains of mice.

[Suspension culture of protocorm-like bodies from the endangered medicinal plant Dendrobium huoshanenese].

OBJECTIVE: To investigate the characteristics of growth, and water-soluble polysaccharide and total alkaloid accumulation in protocom-like bodies (PLBs) of Dendrobium huoshanenese in liquid culture system. METHOD: PLBs were suspended in liquid medium and growth kinetics was analyzed. Water-soluble polysaccharide and total alkaloid content in PLBs were determined by colorimetry. RESULT: PLBs were induced from stem explants of D. huoshanenese regenerants cultured on MS medium supplemented with NAA or NAA and KT at different concentrations. Basal MS medium was suitable for propagation of PLBs. When PLBs were suspended in liquid medium, mumax was 0.044.d-1, t(d) was 15.8 d and the optimum growth time was 4 weeks. Water-soluble polysaccharide and total alkaloid contents in PLBs were 3.75% and 0.0261%, respectively. CONCLUSION: PLBs in liquid culture system show a potential for rapid growth and high metabolite synthesis, which provides possibility for exploiting resources of D. huoshanenese by large scale culture of PLBs.

[Callus formation and its isoflavonoid accumulation in Maackia amurensis].

OBJECTIVE: To obtain Maackia amurensis callus and investigate some factors influencing total isoflavonoid production in callus. METHOD: Different media were used for callus induction from cotyledonary and hypocotyl explants. Total isoflavonoid content was determined by UV spectrophotometer. RESULT: Medium types and hormone combinations influenced both callus formation and their developmental states. MS medium supplemented with NAA/BA or containing 2,4-D was suitable for callus induction from cotyledonary explants. Among the basal media tested, MS, N6 and B5 were suitable for cotyledonary callus formation and SH for hypocotyl callus formation. The formed callus could synthesize isoflavones. Media containing NAA were suitable for isoflavonoid production in cotyledonary callus and media containing 2,4-D/BA were suitable for hypocotyl callus, but 2,4-D/KT at high concentration inhibited the isoflavonoid accumulation in hypocotyl and coytledonary calli. According to the developmentally morphological characters, the formed callus could be classified into four types. II-type callus with yellow in color and friability in texture showed the highest accumulation of isoflavones. CONCLUSION: The influences of medium type and hormone combinations on Maackia amurensis callus formation are reflected in both developmentally morphological characters and isoflavonoid accumulation of calli. Yellow, friable callus induced from cotyledonary and hypocotyl explants in MS or N6 medium supplemented with 2,4-D and BA exhibit the optimum growth and isoflavonoid production.