New Biotransformation Mode of Zearalenone Identified in Bacillus subtilis Y816 Revealing a Novel ZEN Conjugate.

An increasing number of Bacillus strains have been identified, and the removal capacity of zearalenone (ZEN) was determined; however, they failed to reveal the detoxification mechanism and transformation product. Here, Bacillus subtilis Y816, which could transform 40 mg/L of ZEN within 7 h of fermentation, was identified and studied. First, the biotransformation products of ZEN and 17-ß-estradiol (E2) were identified as ZEN-14-phosphate and E2-3-phosphate by HPLC-TOF-MS and NMR, respectively. An intracellular zearalenone phosphotransferase (ZPH) was found through transcriptome sequencing analysis of B. subtilis Y816. The phosphorylated reaction conditions of ZEN by ZPH were further revealed in this work. Furthermore, the phosphorylated conjugates showed reduced estrogenic toxicity compared with their original substances (ZEN and α/ß-zearalenol) using an engineered yeast biosensor system. The first report on the phosphorylated conjugated mode of ZEN in B. subtilis Y816 will inspire new perspectives on the biotransformation of ZEN in Bacillus strains.

Evaluation of different surgical methods in treating recurrent patella dislocation after three-dimensional reconstruction.

PURPOSE: Recurrent patella dislocation (RPD) is the most common complication of patellar instability. The effects of different surgical techniques on the outcome of RPD treatments remain unclear. This study was conducted to compare the effects of three surgical techniques in treating RPD by three-dimensional (3D) reconstruction from computed tomography (CT) scans. METHODS: Sixty-eight patients with RPD and no previous surgical treatments who attended our hospital between October 2010 and 2013 were enrolled and randomly assigned into three groups: (1) medial patellofemoral ligament (MPFL) reconstruction and medial patellar retinaculum (MPR) plication group; (2) lateral patellar retinaculum (LPR) release and MPR plication group; and (3) MPFL reconstruction and LPR release group. Knee joints with flexion of 20° were scanned by a 64-row CT scanner and 3D reconstructed. Congruence angle (CA), patellar tilt angle (PTA), lateral patellofemoral angle (LPFA), and congruence of the lateral patellofemoral articular surface were measured. Knee joint function was evaluated by the Lysholm knee scoring scale, Kujala patellofemoral score, and International Knee Documentation Committee (IKDC) score. RESULTS: Pre-operative clinical characteristics were similar across groups. After treatment, the CA, PTA, and LPFA were reverted to normal post-operatively without statistically significant between-group differences. The MPFL reconstruction and LPR release group had the highest congruence of the lateral patellofemoral articular surface; while the (LPR) release and (MPR) plication group had the lowest Lysholm knee scoring scale, Kujala patellofemoral score, and IKDC score after surgery. CONCLUSIONS: The MPFL reconstruction and LPR release group had the best clinical outcomes among the three surgical methods, as indicated by better joint congruence after 3D joint reconstruction and higher knee function scores.

Common variants of APOE are associated with anti-epileptic drugs resistance in Han Chinese patients.

Purpose/aim of the study: Apolipoprotein E (APOE) has been implicated as one of the susceptibility genes for some subtypes of epilepsy and may be related to anti-epileptic drugs resistance. The purpose of this study was to investigate the possible association between APOE variants and the anti-epileptic drugs resistance in Chinese population. MATERIALS AND METHODS: APOE gene rs429358 and rs7412 variants were genotyped for ϵ2, ϵ3, ϵ4 alleles using amplification refractory mutation system in 480 subjects including 207 anti-epileptic drugs-resistant patients and 273 drug-responsive patients. RESULTS: We found that the frequency of APOE gene rs429358 C allele in the drug resistant patients is higher than that in the drug-responsive patients (14.98% vs. 10.1%, OR = 1.25[1.02 - 1.52], p = 0.017). Moreover, according to the two variants, we analyzed the distributions of -ϵ4 and +ϵ4 alleles of APOE gene and found that there were higher frequencies of +ϵ4 allele in drug-resistant epileptic patients than that in drug-responsive patients (31.8% vs. 13.2%, OR = 1.15[1.05 - 1.25], p = 0.002). CONCLUSIONS: Our study demonstrated that APOE rs429358 variant C allele and ϵ4 allele were associated with the anti-epileptic drugs resistance in Han Chinese patients.

Neurotization of free gracilis transfer with the brachialis branch of the musculocutaneous nerve to restore finger and thumb flexion in lower trunk brachial plexus injury: an anatomical study and case report.

OBJECTIVE: To investigate the feasibility of using free gracilis muscle transfer along with the brachialis muscle branch of the musculocutaneous nerve to restore finger and thumb flexion in lower trunk brachial plexus injury according to an anatomical study and a case report. METHODS: Thirty formalin-fixed upper extremities from 15 adult cadavers were used in this study. The distance from the point at which the brachialis muscle branch of the musculocutaneous nerve originates to the midpoint of the humeral condylar was measured, as well as the length, diameter, course and branch type of the brachialis muscle branch of the musculocutaneous nerve. An 18-year-old male who sustained an injury to the left brachial plexus underwent free gracilis transfer using the brachialis muscle branch of the musculocutaneous nerve as the donor nerve to restore finger and thumb flexion. Elbow flexion power and hand grip strength were recorded according to British Medical Research Council standards. Postoperative measures of the total active motion of the fingers were obtained monthly. RESULTS: The mean length and diameter of the brachialis muscle branch of the musculocutaneous nerve were 52.66±6.45 and 1.39±0.09 mm, respectively, and three branching types were observed. For the patient, the first gracilis contraction occurred during the 4th month. A noticeable improvement was observed in digit flexion one year later; the muscle power was M4, and the total active motion of the fingers was 209°. CONCLUSIONS: Repairing injury to the lower trunk of the brachial plexus by transferring the brachialis muscle branch of the musculocutaneous nerve to the anterior branch of the obturator nerve using a tension-free direct suture is technically feasible, and the clinical outcome was satisfactory in a single surgical patient.

Neurotization of free gracilis transfer with the brachialis branch of the musculocutaneous nerve to restore finger and thumb flexion in lower trunk brachial plexus injury: an anatomical study and case report

OBJECTIVE: To investigate the feasibility of using free gracilis muscle transfer along with the brachialis muscle branch of the musculocutaneous nerve to restore finger and thumb flexion in lower trunk brachial plexus injury according to an anatomical study and a case report. METHODS: Thirty formalin-fixed upper extremities from 15 adult cadavers were used in this study. The distance from the point at which the brachialis muscle branch of the musculocutaneous nerve originates to the midpoint of the humeral condylar was measured, as well as the length, diameter, course and branch type of the brachialis muscle branch of the musculocutaneous nerve. An 18-year-old male who sustained an injury to the left brachial plexus underwent free gracilis transfer using the brachialis muscle branch of the musculocutaneous nerve as the donor nerve to restore finger and thumb flexion. Elbow flexion power and hand grip strength were recorded according to British Medical Research Council standards. Postoperative measures of the total active motion of the fingers were obtained monthly. RESULTS: The mean length and diameter of the brachialis muscle branch of the musculocutaneous nerve were 52.66±6.45 and 1.39±0.09 mm, respectively, and three branching types were observed. For the patient, the first gracilis contraction occurred during the 4th month. A noticeable improvement was observed in digit flexion one year later; the muscle power was M4, and the total active motion of the fingers was 209°. CONCLUSIONS: Repairing injury to the lower trunk of the brachial plexus by transferring the brachialis muscle branch of the musculocutaneous nerve to the anterior branch of the obturator nerve using a tension-free direct suture is technically feasible, and the clinical outcome was satisfactory in a single surgical patient.

Functioning free gracilis transfer to reconstruct elbow flexion and quality of life in global brachial plexus injured patients.

In the study, the functional recovery and relative comprehensive quality of life of cases of global brachial plexus treated with free functioning muscle transfers were investigated. Patients who received functioning gracilis muscle transfer between August 1999 and October 2014 to reconstruct elbow flexion, wrist and fingers extension were recruited. The mean age of the patients was 26.36 (range, 16-42) years. The mean period of time from gracilis transfer to the last follow-up was 54.5 months (range, 12-185 months). Muscle power, active range of motion of the elbow flexion, wrist extension, and total active fingers extension were recorded. SDS, SAS and DASH questionnaires were given to estimate patients' quality of life. 35.71% reported good elbow flexion and 50.00% reported excellent elbow flexion. The average ROM of the elbow flexion was 106.5° (range, 0-142°) and was 17.00° (range, 0-72°) for wrist extension. The average DASH score was 51.14 (range, 17.5-90.8). The prevalence of anxiety and depression were 42.86% and 45.24%. Thrombosis and bowstringing were the most common short and long-term complications. Based on these findings, free gracilis transfer using accessory nerve as donor nerve is a satisfactory treatment to reconstruct the elbow flexion and wrist extension in global-brachial-plexus-injured patients.

Donor-side morbidity after contralateral C-7 nerve transfer: results at a minimum of 6 months after surgery.

OBJECT Donor-side morbidity associated with contralateral C-7 (CC7) nerve transfer remains controversial. The purpose of this study was to evaluate functional deficits in the donor limb resulting from prespinal route CC7 nerve transfer. METHODS A total of 63 patients were included. Forty-one patients had undergone CC7 nerve transfer surgery at least 6 months previously and were assigned to one of 2 groups based on the duration of postoperative follow-up. Group 1 (n = 21) consisted of patients who had undergone surgery between 6 months and 2 years previously, and Group 2 (n = 20) consisted of patients who had undergone surgery more than 2 years previously. An additional 22 patients who underwent CC7 nerve transfer surgery later than those in Groups 1 and 2 were included as a control group (Group 3). Results of preoperative testing in these patients and postoperative testing in Groups 1 and 2 were compared. Testing included subjective assessments and objective examinations. An additional 3 patients had undergone surgery more than 6 months previously but had severe motor weakness and were therefore evaluated separately; these 3 patients were not included in any of the study groups. RESULTS The revised Short-Form McGill Pain Questionnaire (SF-MPQ-2) was the only subjective test that showed a significant difference between Group 3 and the other 2 groups, while no significant differences were found in objective sensory, motor, or dexterity outcomes. The interval from injury to surgery for patients with a normal SF-MPQ-2 score in Groups 1 and 2 was significantly less than for those with abnormal SF-MFQ-2 scores (2.4 ± 1.1 months vs 4.6 ± 2.9 months, p = 0.002). The 3 patients with obvious motor weakness showed a tendency to gradually recover. CONCLUSIONS Although some patients suffered from long-term sensory disturbances, resection of the C-7 nerve had little effect on the function of the donor limb. Shortening preoperative delay time can improve sensory recovery of the donor limb.

[Evaluation of efficiency of traps baited with frequency trembler grid lamps and trap plants for control Heortia vitessoides infectwed in Aquilaria sinensis].

Heortia vitessoides has been a serious defoliating pest of Aquilaria sinensis forests in recent years.The adults displayed strong tropism to the frequency trembler grid lamps and the nectar source plants.The favorite nectar source plants of H.vitessoides adults as the trap plants and the frequency trembler grid lamps in the integrated management of H.vitessoides were studied in the adult eclosion period through both the laboratory and field.The results showed that Kuhnia rosmarnifolia and Santalum album plants showed strong attraction to the H.vitessoides adults, with significant differences among the different nectar source plants.K.rosmarnifolia and S.album as trap plants with board type of planting area to total planting area of 5%-10%, and the frequency trembler grid lamps trapped significantly more adults of H.vitessoides. These results suggested that the frequency trembler grid lamps and trap plants could play an important role in the integrated management of the pest H.vitessoides of A.sinensis.

Diagnostic accuracy of three sensory tests for diagnosis of sensory disturbances.

BACKGROUND: Clinical diagnosis of sensory disturbances is extremely challenging, partly because the utility of sensory tests is questionable. Transection of C7 nerve root provides an objective assessment model to determine the diagnostic accuracy of sensory tests. The purpose of this study was to investigate the diagnostic accuracy of Semmes-Weinstein Monofilaments (SWM; Sammons Preston, Bolingbrook, IL), static two-point discrimination (s2PD), and current perception threshold (CPT) tests in patients with contralateral C7 nerve root transfer. METHODS: The contralateral index finger of patients was tested preoperatively and 6 weeks postoperatively, including the Revised Short-Form McGill Pain Questionnaire (SF-MPQ-2), SWM, s2PD, and CPT tests. The SF-MPQ-2 was used as a gold standard for sensory disturbances. RESULTS: A total of 38 patients were included in the study. The likelihood probability ratio of SWM and CPT tests generated small shifts in probability, whereas the s2PD test did not show any capacity to detect sensory disturbances. The areas under the curves (AUC) for SWM and CPT tests were 0.724 and 0.697, respectively. Based on different positive test standards, the AUCs for s2PD (≥ 6 mm) and s2PD (≥ 7 mm) tests were 0.632 and 0.658, respectively. When CPT and SWM tests were combined as a measure, the AUC increased to 0.763. CONCLUSION: The SWM and CPT tests show a small capacity to detect sensory disturbances, whereas the s2PD test does not show any diagnostic capacity. To better understand sensory disturbances, a comprehensive testing protocol including valid measures of physical impairment and symptom-specific measurement tools should be adopted.

A water-alcohol extract of Citrus grandis whole fruits has beneficial metabolic effects in the obese Zucker rats fed with high fat/high cholesterol diet.

Epidemiological studies suggest that citrus fruits and compounds such as flavonoids, limonoids and pectins have health promoting effects. Our aim was to study the effects of Citrus grandis (L.) Osbeck var. tomentosa hort. fruit extract on the energy metabolism. A whole fruit powder from dry water and alcohol extracts of C. grandis containing 19% naringin flavonoid was prepared. The effects of the citrus extract were followed in the obese Zucker rats fed with the HFD. The circulatory levels of GLP-1 decreased significantly by the extract in comparison to the HFD group, whereas the decreased ghrelin levels were reversed. The levels of PYY were decreased in all HFD groups. The leptin amounts decreased but not significantly whereas insulin and amylin were unchanged. The cholesterol and glucose levels were somewhat but not systematically improved in the HFD fed rats. Further studies are needed to identify the active compounds and their mechanisms.

[Screening for virulence strains of Metarhizium against Dorysthenes hydropicus pascoes].

OBJECTIVE: The aim of the present study was to screen the Metarhizium strains with high virulence against the larvae of Dorysthenes hydropicus, a serious pest of Citrus grandis. METHOD: Thirty six strains of Metarhiziums were isolated from the soil of C. grandis GAP base and collected from other institutions, and the pathogenicity of these strains against 1st instar larvae of D. hydropicus was detected at concentration of 1 x 10(8) conidia/g. The high violence strains against D. hydropicus were cultivated in sabouraud dextrose yeast medium at first, then transfer to rice grain. And the sporulations of these violent strains against D. hydropicus were detected. RESULT: Twenty-eight strains showed virulence against D. hydropicus by preliminary study, and 7 strains of them were collected for further study, 6 of the 7 showed high virulence, the highest cadaver rate was higher than 74%. The conidia production of strain 1 and strain 4 were 2.35 +/- 0. 25 (1 x 10(9) conidia/g), 2.21 +/- 0.27 (1 x 10(9) conidia/g), respectively, showed significantly higher than other strains. CONCLUSION: Strain 1 and strain 4 of the 36 Metarhiziums strains showed high virulence against D. hydropicus, and the highest sporulation ability, so they have a best application prospect.

[UPLC-Q-TOF/MS analysis of naringin and naringenin and its metabolites in rat urine and feces after intragastric administration of alcohol extract of Exocarpium Citri grandis].

To analyze naringin, naringenin and its metabolites in rat urine and feces after intragastric administration of alcohol extract of Exocarpium Citri Grandis, healthy SD rats were fed with alcohol extract of Exocarpium Citri Grandis for 3 days. On the last day, 0-24 h feces and 0-4 h, 4-8 h, 8-24 h urine were collected and analyzed by UPLC-Q-TOF/MS. The post-acquisition data were processed using Metabolynx The result is that naringin and its 6 metabolites, naringenin and its 4 metabolites were detected in the urine of rat. Meanwhile, naringin and its 3 metabolites, naringenin and its 2 metabolites were detected in the feces of rat.

[Primary studies on biological characteristics of Dorysthenes hydropicus].

OBJECTIVE: To study the biological characteristics of Dorysthenes hydropicus in the farm of Cirtus grandis, and offer scientific evidence for prevention and controlling of D. hydropicus. METHOD: Indoor-rearing and light trap were applied to study the biological characteristics, development course and harmful effect of D. hydropicus. RESULT: D. hydropicus reproduces one generation in 1-2 year in Guangdong province, and overwinters in the form of larvae. Its imago comes out of the earth mainly in late May after mature. The body length has great individual diversity normally ranged from 25-60 cm, It also shows strong phototaxy. One lamp can trap more than 2 000 of them per night. Female imago has a large egg load with the maximum amount of 543. The eggs hatching is in depth of 1-3 cm soil. The dominant hatching period of egg is from late June to early July, and hatchability is over 85%. The living space of larva ranges from 15-60 cm in soil. D. hydropicus has caused serious harm and lead to thousands of Cirtus grandis trees death every year. CONCLUSION: Dorysthenes hydropicus showed serious threat to the growth of Cirtus grandis and should be prevented and controlled.

Bioassay of estrogenic activity of effluent and influent in a farm wastewater treatment plant using an in vitro recombinant assay with yeast cells.

OBJECTIVE: Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on industrial discharges, while the impact of livestock wastes as a source of endocrine disrupters in aquatic environments has been rarely elucidated. In order to investigate the contribution of environmental estrogens from livestock, the estrogenic activity in water samples from a farm wastewater treatment plant was analyzed by a recombinant yeast screening method. METHODS: The extracts prepared from 15 selected water samples from the farm wastewater treatment plant, among which 6 samples were from pre-treatment section (influents) and 9 from post-treatment section (effluents), were analyzed for estrogenic activity by cellar bioassay. Yeast cells transfected with the expression plasmid of human estrogen receptor and the Lac Z reporter plasmid encoding beta-galactossidase, were used to measure the estrogen-like compounds in the farm wastewater treatment plant. RESULTS: The wastewater samples from influents showed a higher estrogenic potency than the effluent samples showing a low induction of beta-galactossidase relative to solvent control condition. By comparison with a standard curve for 17 beta-estradiol (E2), estrogenic potency in water samples from the influents was calculated as E2-equivalent and ranged from 0.1 to 150 pM E2-equivalent. The estrogenic potency in water samples from the effluents was significantly lower than that in the influents, and 7 water samples had less detectable limit in the total of 9 samples. CONCLUSION: Yeast bioassay of estrogenic activity in most of the samples from the farm wastewater after disposal by traditional sewage treatment showed negative results.

[Establishment of a transgenic cell model for preliminary screening of chemopreventive agents].

To develop a GFP transgenic cell model under the transcriptional control of TK promoter adjacent to which ARE enhancer was inserted. Synthetic oligonucleotide ARE motif was annealed and purified then inserted into pTK-GFP to construct the vector of pARE-TK-GFP. The TK and ARE-TK fragments were amplified by PCR and cloned into pEGFP-N1 to reconstruct eukaryotic expression vectors of pTK-GFP/Neo and pARE-TK-GFP/Neo. They were transfected into HepG2 cells and clones resistant G418 were isolated. PDTC and Lentinan were used to induce the cell levels of GFP and the fluorescence was measured using a fluorescence plate reader. The results showed that the induced level of GFP is significantly increased and have dose-dependeny in a certain range. This findings indicated that such a cell model offered a potential platform for preliminary screening of all kinds of natural or synthetic chemopreventive agents.

[Use W303-1A/hER-ERE-Lac Z to determine estrogenic compounds in traditional Chinese materia medica].

OBJECTIVE: To study the content of phytoestrogen in dissimilarity herbs. METHOD: The activity of phytoestrogen in heat-clearing drugs, drugs for relieving exterior syndrome, diuretic, anastaltics, tonics and astringents were detected based on the recombinant yeast cell (W303-1A/hER-ERE-Lac Z). The estrogenic activity in traditional Chinese materia medica were assayed quantitatively by determining the expression of beta-galactosidase. RESULT: The phytoestrogen concentration (6.35 x 10(-3) nmol x g(-1) E2 equivalent) in heat-clearing drugs was the highest while that in anastaltic and tonic drugs was the lowest, which was less than the detected limit. CONCLUSION: Compared with the other traditional Chinese materia medica, the content of phytoestrogen, which can bind to estrogen receptor, in giant knotweed rhizome, forsythia suspense, ash bark, baical skullcap root and ophiopogonis tuber were higher.

Testing potential effect of environmental endocrine disruptors in cow milk on reproductive index in female rats.

OBJECTIVE: To study the effect of endocrine disruptor chemicals in cow milk on female reproductive system. METHODS: A two-generation reproduction was conducted according to U. S. FDA standard. Milk was fed in special bottle to Wistar rats of both sexes through two successive generations (F0 and F1) in the milk group while artificial milk was fed to rats in the control group. Twenty-four rats of each sex were mated in each group. Measurements were made according to this guideline. RESULTS: Reproductive parameters in the milk group such as fertility index, gestation index, weights of uterus and ovary, days of vaginal opening, estrous cycles, histological morphological changes were comparable to those in the control group. However, the means of body weight had some differences. The body weight gains increased significantly in the milk-treated group in F1 and F2 generation compared with those in the control group. The concentration of insulin-like growth factor-1 (IGF-1) in blood in the milk group was comparable to that in the control group, but the standard deviation changed greatly in the milk-treated rats. CONCLUSION: Endocrine disruptor chemicals in milk have no severe effects on the female reproductive system.

A C-type lectin-like protein from Agkistrodon acutus venom binds to both platelet glycoprotein Ib and coagulation factor IX/factor X.

Agkisacutacin, a C-type lectin-like protein (CLP) isolated from Agkistrodon acutus venom, had been previously identified as an antagonist of platelet aggregation induced by ristocetin, as well as a certain extent fibrinogenlytic activity. In this study, agkisacutacin was further purified by three-step chromatography, and its biological functions were characterized. Agkisacutacin after further purification retained the effect on ristocetin-induced, von Willebrand factor-dependent platelet aggregation, while it lost the fibrinogenlytic activity. FACS and ELISA assays showed that agkisacutacin belongs to membrane glycoprotein Ib-binding protein (GPIb-bp) for it could block and inhibit the binding of anti-GPIb antibody to GPIb. Especially, agkisacutacin exhibits anti-coagulant activity and the function as IX/X-binding protein was confirmed by PAGE and ELISA. So, agkisacutacin is the first reported CLP that binds to both platelet membrane receptors and coagulation factors.

[The effects of estrogen-like products in milk on prostate and testes].

OBJECTIVES: To study the effects of estrogen-like products in milk on the male reproductive system. METHODS: The incidence rates of testicular and prostatic cancers in 42 countries (regions), collected from the cancer incidence in five continent database (1988-1992), were correlated with the relative food consumption in these countries, which was provided by FAO database (1961-1990); two-generation reproduction test was taken by using Wistar rats according to FDA standard. RESULTS: Among the food items examined, milk was most closely correlated (r = 0.711) with prostatic cancer incidence, followed by meat and coffee. As for testicular cancer, cheese was most closely correlated (r = 0.804) with the incidences in ages of 20-39, followed by animal fat and milk. No significant decreases of reproductive organ weight were found after the rats had been fed a lot of milk for long time. CONCLUSIONS: The results suggest that further studies should be taken on the effects of estrogen-like products in milk on the male reproductive health.