Genomic and structural basis for evolution of tropane alkaloid biosynthesis.

The tropane alkaloids (TAs) cocaine and hyoscyamine have been used medicinally for thousands of years. To understand the evolutionary origins and trajectories of serial biosynthetic enzymes of TAs and especially the characteristic tropane skeletons, we generated the chromosome-level genome assemblies of cocaine-producing Erythroxylum novogranatense (Erythroxylaceae, rosids clade) and hyoscyamine-producing Anisodus acutangulus (Solanaceae, asterids clade). Comparative genomic and phylogenetic analysis suggested that the lack of spermidine synthase/N-methyltransferase (EnSPMT1) in ancestral asterids species contributed to the divergence of polyamine (spermidine or putrescine) methylation in cocaine and hyoscyamine biosynthesis. Molecular docking analysis and key site mutation experiments suggested that ecgonone synthases CYP81AN15 and CYP82M3 adopt different active-site architectures to biosynthesize the same product ecgonone from the same substrate in Erythroxylaceae and Solanaceae. Further synteny analysis showed different evolutionary origins and trajectories of CYP81AN15 and CYP82M3, particularly the emergence of CYP81AN15 through the neofunctionalization of ancient tandem duplication genes. The combination of structural biology and comparative genomic analysis revealed that ecgonone methyltransferase, which is responsible for the biosynthesis of characteristic 2-substituted carboxymethyl group in cocaine, evolved from the tandem copies of salicylic acid methyltransferase by the mutations of critical E216 and S153 residues. Overall, we provided strong evidence for the independent origins of serial TA biosynthetic enzymes on the genomic and structural level, underlying the chemotypic convergence of TAs in phylogenetically distant species.

Lactate dehydrogenase as promising marker for prognosis of brain metastasis.

BACKGROUND: Lactate dehydrogenase (LDH) is a biomarker for cancer. However, the relationship between serum LDH levels and the survival of patients with brain metastasis has been fully revealed. We aimed to evaluate the serum LDH levels and assess its prognostic value in patients with BM. METHODS: The serum LDH levels were collected from 2507 patients with BM. Patients were categorized into four groups according to the quartile of serum LDH levels. The association between serum LDH levels and overall survival (OS) was evaluated using Cox regression models and Kaplan-Meier curves. Three predictive models were used to evaluate patients. RESULTS: The Kaplan-Meier curve for survival by the serum LDH group demonstrates clear separation between four groups (P < 0.001). The participants in the lower group had longer OS than those in the higher group. After adjusting in multivariate Cox regression models remained significant for patients in the Q4 compared with patients in the Q1 (Q4:Q1 OR 1.58, 95% CI 1.38-1.80). Furthermore, the GPA-LDH model generates a pooled area under the curve of 0.630 (95% CI 0.600, 0.660). CONCLUSIONS: Serum LDH levels and OS in patients with brain metastasis is an inverse association. Moreover, Serum LDH levels can improve the prognosis of the GPA model.

Effects of immersing treatment of curcumin and piperine combined with vacuum packaging on the quality of salmon (Salmo salar) during cold chain logistics.

In order to study the effects of the compound preservatives (curcumin and piperine (CP)) and vacuum packaging (VP) on the quality of salmon during cold chain logistics suffered from temperature abuse, the physiochemical indexes (texture, water holding capacity (WHC), total volatile basic nitrogen (TVB-N), thiobarbituric acid reactive substances (TBARS), free amino acids (FAA) contents), microbial indicators (total mesophilic bacteria count (MBC), total psychrotrophic bacteria count (PBC), H2S-producing bacteria count (HSBC)) were determined, and the moisture changes were explored by near-infrared (NIR) spectroscopy and low-field nuclear magnetic resonance (LF-NMR). The results showed that the treatment of curcumin and piperine in combination with vacuum packaging could maintain the quality of salmon suffered from temperature abuse most effectively. At the end of storage, the MBC of VP+CP was only 4.95 log CFU/g, which was about 1 log CFU/g lower than the control sample stored at the same condition. The combined treatment also retarded the increase of TVB-N, TBARS, and the decrease of hardness, springiness, and a* value, as well as water migration in salmon, contributing to higher water holding capacity and better appearance. Besides, VP+CP retarded the decrease of free glutamate, which contributed to umami taste. Due to the biological activity and safety of the preserves, the combined treatment could be a promising method for preservation of seafood.

[Clinical observation of highly myopic eyes with retinoschisis after phacoemulsification].

OBJECTIVE: To evaluate the changes of the retinal thickness in the highly myopic eyes with retinoschisis after phacoemulsification by optical coherence tomography (OCT) and to analyze their postoperative visual acuity. METHODS: It was a retrospective case series study. This study included 25 (35 eyes) highly myopic patients with retinoschisis. One week, 1 and 3 months after phacoemulsification, the best corrected visual acuity (BCVA) was measured and OCT was performed to evaluate the central foveal thickness (CFT) as well as the maximum neurosensory thickness (MNT). These 35 eyes were divided into low and high accumulated energy complex parameter (AECP) groups, and ≤ 28.00 mm, > 28.00 and ≤ 30.00 mm, > 30.00 mm groups according to the axial length respectively. Variance analysis of data obtained from repeated measurement, Wilcoxon rank sum test, linear correlation and one-way ANOVA were used to analyze the data. RESULTS: According to the location of schisis, the 35 eyes were classified as foveal schisis (20 eyes) and nonfoveal schisis (15 eyes). Among the 35 eyes, there were 30 eyes with outer retinoschisis and 5 with compound schisis. The mean BCVA (logMAR) preoperatively and 1 week, 1 and 3 months postoperatively was 0.23 ± 0.24, 0.59 ± 0.38, 0.57 ± 0.38 and 0.60 ± 0.36 respectively (F = 31.15, P = 0.000). CFT preoperatively and 1 week, 1 and 3 months postoperatively was (255.46 ± 197.57) µm, (256.80 ± 199.88) µm, (274.37 ± 246.87) µm, and (268.60 ± 238.41) µm (F = 0.99, P = 0.420); while MNT was (473.31 ± 175.52) µm, (477.71 ± 188.71) µm, (486.60 ± 229.17) µm and (482.63 ± 208.82) µm respectively (F = 0.26, P = 0.857). BCVA in eyes with foveoschisis and in eyes with nonfoveal schisis was statistically significantly different at 1 week, 1 and 3 months postoperatively (Z = -0.580, P = 0.048; Z = -2.147, P = 0.030 and Z = -2.099, P = 0.034). BCVA (0.43 ± 0.28) was inversely correlated with CFT (497.42 ± 281.49) µm in the eyes with foveoschisis unaccompanied with a macular lamellar hole at 3 months postoperatively (r = -0.667, P = 0.018). There were no significant differences in the changes of the thickness of the retina between the groups with different AECP or among the groups with different axial lengths (Z = -0.314 - 1.290, P = 0.192−0.741 and F = 0.15−0.62, P = 0.545−0.859). CONCLUSIONS: Cataract surgery has no significant effect on thickness of the retina in the highly myopic eyes with retinoschisis. The patients with retinoschisis could achieve good visual recovery after phacoemulsification. Postoperative visual acuity is related to the location and degree of retinoschisis. Besides, patients with non-foveal schisis could obtain better visual acuity than those with foveoschisis. Further investigation on the development of retinoschisis and the complications after surgery is required.

Immediate precision of the digital osteotomy template in the digital stackable template: a clinical study.

OBJECTIVES: This study aimed to evaluate the immediate accuracy of the digital osteotomy template in the digital stackable template. METHODS: From November 2018 to January 2020, 4 patients with dentition loss were selected from the Prosthodontics Department, West China Stomatological Hospital. All patients met the conditions for immediate planting and immediate restoration. Owing to the lack of vertical target-restoration space, the implantation plan included intraoperative osteotomy. According to the preoperative cone beam CT (CBCT) data, combined with aesthetic digital smile design (DSD) analysis, virtual wax design, and so on, the ideal bone plane design was performed. According to the virtual osteotomy plane, the virtual implantation plan was designed, and then the digital stackable template assuming osteotomy template, implantation guide, and temporary restoration were made and 3D printed. Osteotomy was performed under the guidance of digital osteotomy template during the operation. The preoperative CBCT and postoperative CBCT of all patients overlapped, the deviation between the actual osteotomy and the ideal osteotomy was calculated, and the angle deviation between the postoperative bone plane and the ideal bone plane was measured. RESULTS: The ave-rage volume deviation between the postoperative design and the ideal one was 492.94 mm³, accounting for 21.21% of the preset osteotomy volume. The average deviation between the postoperative osteotomy and the ideal osteotomy in four patients was 0.024 8 mm. The average angle between the postoperative bone plane and the ideal bone plane was 6.03°. CONCLUSIONS: The displacement deviation of virtual osteotomy design and the actual osteotomy one under the guidance of digital osteotomy template in the digital stackable template are highly consistent with the original design. Thus, this clinical technique is worth popularizing, accurate, and quantifiable.

Clinical outcomes and 5-year follow-up results of keratosis pilaris treated by a high concentration of glycolic acid.

BACKGROUND: Keratosis pilaris is a hereditary abnormal keratosis of the hair follicle orifice. Gray-brown keratotic plugs in the pores and dark red keratotic papules at the openings of hair follicles can be seen, which contain coiled hair and are often accompanied by perifollicular erythema and pigmentation. Glycolic acid can correct the abnormalities of hair follicular duct keratosis and eliminate excessive accumulation of keratinocytes. It also promotes skin metabolism and accelerates the melanin metabolism. The therapeutic effect is related to the glycolic acid concentration. AIM: To evaluate the efficacy and safety of a high concentration of glycolic acid in the treatment of keratosis pilaris, and to observe the outcomes at 5-year of follow-up. METHODS: Twenty-five participants were recruited and areas with typical keratosis pilaris were selected as testing sites. High concentrations of glycolic acid (50% or 70%) were applied to a circular area (d = 8 cm, S = 50 cm2) and repeated four times, on days 0, 20, 40 and 60. Before each treatment and 20 d after the last treatment, on days 0, 20, 40, 60, and 80 and at a 5-year follow-up, The number of follicular keratotic papules were counted and the extent of perifollicular erythema and pigmentation was determined. At the same time, the participants provided subjective evaluations of treatment efficacy and safety. RESULTS: Treatment effectiveness was indicated by the percentage of keratotic papules in the test site, on days 20, 40, 60 and 80, which were 8%, 12%, 36%, and 60%, respectively. Compared with day 0, each difference was significant (P < 0.05). Compared with day 0, differences in melanin content (M) in the skin and skin lightness (L) on days 40, 60 and 80, the were statistically significant (P < 0.05); skin hemoglobin content (E) on days 60 and 80 was statistically different as compared with before treatment (P < 0.05). There were no significant differences in the number of keratotic papules, M, L, and E in 9 participants at the 5-year follow-up compared with before treatment (P > 0.05%). CONCLUSION: A high concentration of glycolic acid significantly improved skin roughness as well as follicular hyperpigmentation of patients with keratosis pilaris. The treatment was relatively safe, but there was no significant difference at the 5-year follow-up compared to before treatment.