Primulaweiliei (Primulaceae), a new species from Hubei, Central China.

In this study, we describe and illustrate a new species, Primulaweiliei L.S.Yang, Z.K.Wu & G.W.Hu, from the Shennongjia Forestry District, Hubei Province in Central China. It is morphologically assigned to Primulasect.Aleuritia based on its dwarf and hairless habit, long petiole, fruits longer than calyx and covered by farina on the scape. This new species is similar to P.gemmifera and P.munroisubsp.yargongensis in the same section, but it can be distinguished by its smaller calyxes, homostylous flowers, corolla tube throat without annular appendage and only 1-2 flowers in each inflorescence. Based on the assessment conducted according to the IUCN Red List criteria, we propose that P.weiliei be classified as a Critically Endangered (CR) species.

Development of polymorphic microsatellite markers for distylous-homostylous Primula secundiflora (Primulaceae) using HiSeq sequencing.

Primula secundiflora is an insect-pollinated, perennial herb belonging to section Proliferae (Primulaceae) and exhibits considerable variation in mating system with predominantly outcrossing populations comprising long-styled and short-styled floral morphs and selfing populations comprising only homostyles. To facilitate future investigations of the population genetics and mating patterns of this species, we developed 25 microsatellite markers from P. secundiflora using next-generation sequencing and measured polymorphism and genetic diversity in a sample of 30 individuals from three natural populations. The markers displayed relatively high polymorphism, with the number of observed alleles per locus ranging from three to 16 (mean = 8.36). The observed and expected heterozygosities ranged from 0.100 to 1.000 and 0.145 to 0.843, respectively. Twenty-one of the loci were also successfully amplified in P. denticulata. These microsatellite markers could provide powerful tools for investigating patterns of population genetic diversity and the evolutionary relationships between distyly and homostyly in this species.

Parallel evolution of morphological and genomic selfing syndromes accompany the breakdown of heterostyly.

Evolutionary transitions from outcrossing to selfing in flowering plants have convergent morphological and genomic signatures and can involve parallel evolution within related lineages. Adaptive evolution of morphological traits is often assumed to evolve faster than nonadaptive features of the genomic selfing syndrome. We investigated phenotypic and genomic changes associated with transitions from distyly to homostyly in the Primula oreodoxa complex. We determined whether the transition to selfing occurred more than once and investigated stages in the evolution of morphological and genomic selfing syndromes using 22 floral traits and both nuclear and plastid genomic data from 25 populations. Two independent transitions were detected representing an earlier and a more recently derived selfing lineage. The older lineage exhibited classic features of the morphological and genomic selfing syndrome. Although features of both selfing syndromes were less developed in the younger selfing lineage, they exhibited parallel development with the older selfing lineage. This finding contrasts with the prediction that some genomic changes should lag behind adaptive changes to morphological traits. Our findings highlight the value of comparative studies on the timing and extent of transitions from outcrossing to selfing between related lineages for investigating the tempo of morphological and molecular evolution.

Comparison of Cerebrospinal Fluid Cellular Analysis by Optical Microscopy and Sysmex XN 9000.

BACKGROUND: This study was designed to compare the body fluid module of Sysmex XN9000 (XN-BF) with optical microscopy (OM) for cerebrospinal fluid (CSF) analysis after two-step cell slide centrifuge (TSCSC), defining the best procedure for CSF optical microscopy analysis. METHODS: Items of RBC, WBC enumeration and differentiation were observed. The cell count and morphologic evaluation of the cellular composition by OM was carried out both with and without two-step cell slide centrifuge (TSCSC) and were compared the data with XN-BF. RESULTS: There were 69.98 ± 4.94 RBC and 36.98 ± 3.39 WBC in one OSCSC microscopic field whereas there were 96.35 ± 5.41 RBC and 66.15 ± 4.85 WBC in one TSCSC microscopic field in the same sample (*200). There was a statistical difference between those two methods (p = 0.000). Excellent correlation was found between total cell count with both OM and XN-BF. The R2 value for RBC and WBC counts were 0.99 and 0.96, respectively. For WBC differential, the R2 values were 0.98 for PMN and 0.70 for MN. Correlation of MN was poorer than PMN. As far as the tumor cell, phagocyte, and plasma cell with high fluorescence were concerned, OM were not consistent with XN-BF. CONCLUSIONS: The TSCSC procedure contributes to the separation of cells and other ingredients. XN-BF displays excellent performance at RBC and WBC cell count except for mononuclear cells, tumor cells, phagocytes, and leukemia cells. which makes it just a practical alternative to total cell (WBC, RBC) count for CSF samples. Detailed morphologic workup of CSF samples is mandated in all cases with meningoencephalitis, elevated cell count, sub-arachnoid hemorrhage and meningeal carcinomatosis, the TSCSC procedure is recommended.

Primulajiaozishanensis (Primulaceae), a new species in Primula sect. Petiolares subsect. Davidii from Yunnan, China.

Primulajiaozishanensis Z.K.Wu, W.H.Yang & Yuan Wu, a new species of Primulaceae is described and illustrated from Jiaozi Snow Mountain of Dongchuan District, Yunnan, China. Morphological evidence supports P.jiaozishanensis as a member of P.sect.Petiolaressubsect.Davidii, which is characterized by firmly papery or leathery leaves, with veins impressed adaxially, often prominently raised and alveolate abaxially. The new species is characterized by having long and stout rhizomes, smaller leaves with short petioles, short or almost obsolete scape, and larger flowers. The distribution, phenology and conservation status of the new species are also provided.

Rediscovery of Primulabrachystoma (Primulaceae), a rare species endemic to Gaoligong Mountain of Chinese-Burma frontier.

The rare Primulabrachystoma W.W.Sm. is an endemic species confined to Gaoligong mountain of Chinese-Burma frontier, which has been rediscovered from the same region after nearly 100 years. In total, 11 specimens from Gaoligong Mountain have been found in the herbaria worldwide, since its first collection in 1920 by Farrer, Reginald John. Previously, this species was described as homostylous but our finding shows the species also exhibited heterostyly. A complete description of the species, the distribution, morphological comparison and identification key from closely related species are provided here. An assessment of its conservation status suggests that the species is 'Endangered' (EN).

Primulapingbaensis (Primulaceae), a new species from Guizhou, China.

Primulapingbaensis Na Zhang, X.Q.Jiang & Z.K.Wu, a new species of Primulaceae from Gaofeng Mountain of Pingba county, Guizhou, China, is described and illustrated. Morphological evidence supports P.pingbaensis as a member of P.sect.Petiolares on account of scape elongating, pedicels conspicuously thickening in fruit, and its capsule cracking irregularly round the top and crumbling away. Amongst the members of subsect. Davidii, the new species is characterized by having a uniquely smooth leaf blade due to inconspicuously raised veinlets and homostylous flowers with the style usually extending beyond the anthers. The distribution, phenology and conservation status of the new species are also provided.

Primula luquanensis sp. nov. (Primulaceae), a New Species from Southwestern China, Reveals a Novel Floral Form in the Heterostyly-Prevailing Genus.

A new species, Primula luquanensis Z.K.Wu and Wei Zhou sp. nov. (Primulaceae) is described and illustrated from Yunnan Province, China. It is morphologically assigned to P. sect Aleuritia based on its dwarf and hairless habit and coverage by farina on both sides of the leaf blade and scape. This new species is similar to P. nutantiflora and P. yunnanensis, but it is easily distinguished by its stolons, solitary bract, bell-shaped corolla and monomorphic floral form. The new species also has a substantially reduced corolla tube, presenting a unique floral form in a genus where heterostyly typically prevails.

Primulalongipilosa (Primulaceae), a new species from Yunnan, China.

Primulalongipilosa from SW Yunnan, China, is described as a species new to science and illustrated. The systematic placement of this new species is also discussed based on an nrITS molecular tree. It is morphologically most similar to P.mollis, but differs from the latter in its racemose inflorescence, green calyx tube, pink to pink rose corolla, stamens at 1/3 length above the base of the corolla tube and applanate globose capsule.

The complete chloroplast genome of Indigofera stachyodes (Fabaceae), a traditional Chinese medicinal plant.

Indigofera stachyodes Lindl. is a traditional medicinal plant in southwestern China. In this study, we report the complete chloroplast genome sequence of I. stachyodes, using next-generation sequencing technology. The complete chloroplast genome of I. stachyodes was 158,039 bp in length with an overall GC content 35.80%, containing a large single-copy (LSC) region of 88,772 bp, a small single-copy (SSC) region of 18,733 bp, and a pair of inverted repeats (IRs) regions of 25,267 bp. In total, there are 128 genes (83 protein-coding genes (PCGs), eight ribosomal RNA (rRNA) genes, and 37 tRNA genes) in the whole chloroplast genome, including 113 unique genes (78 unique PCGs, 31 unique tRNAs, and four unique rRNAs). The phylogenetic analysis indicated that I. stachyodes formed a monophyletic clade with I. tinctoria and I. linifolia, showing that they have close relationship. The complete chloroplast genome of I. stachyodes provides valuable genomic information for the phylogeny, molecular identification and sustainable utilization of this species.

The complete chloroplast genome of Periploca forrestii (Apocynaceae), a traditional Chinese medicinal plant.

Periploca forrestii Schltr. is a traditional medicine plant in southwestern China. In this study, we characterize the complete chloroplast (cp) genome of P. forrestii based on next-generation sequencing. The cp genome is 154,140 bp in size with an overall GC content 38.2%, including a large single-copy (LSC) region (84,941 bp), a small single-copy (SSC) region of 17,619 bp, and two inverted repeats (IRs) regions, each of 25,790 bp. A total of 130 genes (85 protein-coding genes, 8 ribosomal RNA (rRNA) genes, and 37 transfer RNA (tRNA genes)) are annotated in the whole chloroplast genome, containing 113 unique genes (79 unique CDSs, 30 unique tRNAs, and 4 unique rRNAs). The phylogenetic analysis indicated that P. forrestii formed a monophyletic clade with the same genus plant P. sepium, showing that they have close relationship. The complete chloroplast genome of P. forrestii provides valuable genomic information for the phylogeny, molecular identification and sustainable utilization of this species.

Hemoglobin Fukuoka caused unexpected hemoglobin A1c results: A case report.

BACKGROUND: Glycated hemoglobin (Hb) (HbA1c) is an indicator that is used to diagnose and monitor the treatment of diabetes. Many factors can affect the detection of HbA1c. One of the most important of these factors is the Hb variant. Here, we report a rare Hb variant and evaluate its effect on HbA1c. CASE SUMMARY: A 35-year-old man was suspected of harboring an Hb variant following the measurement of HbA1c with the Variant II Turbo 2.0 Hb detection system during a routine examination. Subsequently, we used the Arkray HA-8160 and ARCHITECT c4000 system to reanalyze HbA1c. Finally, the Hb variant was detected with a Capillary2FP analyzer that operates on the principle of capillary electrophoresis. We also used gene sequencing to investigate the mutation site. The value of HbA1c detected with the Variant II Turbo 2.0 system was 52.7%. However, the Arkray HA-8160 system did not display a result while the ARCHITECT c16000 system showed a result of 5.4%. The Capillary2FP analyzer did not reveal any abnormal Hb zones. However, gene sequencing identified the presence of a mutation in the Hb ß2 chain [CD2(CAC>TAC), His>Tyr, HBB: c.7C>T]; the genotype was Hb Fukuoka. CONCLUSION: Hb variants could cause abnormal HbA1c results. For patients with Hb variants, different methods should be used to detect HbA1c.

The complete chloroplast genome of a distylous-homostylous species, Primula homogama (Primulaceae).

Primula homogama F. H. Chen & C. M. Hu (Primulaceae) is endemic to the Emei Mountain of China. In this study, we characterized the complete chloroplast genome of P. homogama based on next-generation sequencing (NGS). The complete chloroplast genome of P. homogama was 154,677 bp in size with a typical quadripartite structure, containing a large single-copy (LSC) region of 85,299 bp and a small single-copy (SSC) region of 17,816 bp. These two regions were separated by a pair of inverted repeat regions (IRs), each of 25,781 bp. A total of 130 functional genes were encoded, consisted of 86 protein-coding genes (PCG), 36 tRNA genes, and eight ribosomal RNA (rRNA) genes.

The Genomic Selfing Syndrome Accompanies the Evolutionary Breakdown of Heterostyly.

The evolutionary transition from outcrossing to selfing can have important genomic consequences. Decreased effective population size and the reduced efficacy of selection are predicted to play an important role in the molecular evolution of the genomes of selfing species. We investigated evidence for molecular signatures of the genomic selfing syndrome using 66 species of Primula including distylous (outcrossing) and derived homostylous (selfing) taxa. We complemented our comparative analysis with a microevolutionary study of P. chungensis, which is polymorphic for mating system and consists of both distylous and homostylous populations. We generated chloroplast and nuclear genomic data sets for distylous, homostylous, and distylous-homostylous species and identified patterns of nonsynonymous to synonymous divergence (dN/dS) and polymorphism (πN/πS) in species or lineages with contrasting mating systems. Our analysis of coding sequence divergence and polymorphism detected strongly reduced genetic diversity and heterozygosity, decreased efficacy of purifying selection, purging of large-effect deleterious mutations, and lower rates of adaptive evolution in samples from homostylous compared with distylous populations, consistent with theoretical expectations of the genomic selfing syndrome. Our results demonstrate that self-fertilization is a major driver of molecular evolutionary processes with genomic signatures of selfing evident in both old and relatively young homostylous populations.

Ceropegia jinshaensis (Apocynaceae), a new species from northwestern Yunnan, China.

Ceropegia jinshaensis D.T.Liu & Z.K.Wu (Asclepiadoideae, Apocynaceae), a new species from northwestern Yunnan along the upper Yangtze river of China, is described and illustrated. This species is similar to C. meleagris H. Huber, C. dorjei C. E. C. Fischer and C. aridicola W. W. Smith, but can be distinguished easily by its leaf shape and floral features, especially the corolla shape and size, the interior of corolla tube and coronal characters.

Primula dongchuanensis (Primulaceae), a new species from northern Yunnan, China.

Primula dongchuanensis Z.K.Wu & Yuan Huang, a new species of Primulaceae from Dongchuan of northern Yunnan, China, is described and illustrated. Both morphological and molecular evidence support P. dongchuanensis as a member of the sect. Proliferae. It is similar to P. aurantiaca W.W.Smith & Forrest, but is distinguished by having unique raceme inflorescences. Its distribution, phenology and conservation status are also provided.

Phylogenomic analysis reveals multiple evolutionary origins of selfing from outcrossing in a lineage of heterostylous plants.

Evolutionary transitions from outcrossing to selfing often occur in heterostylous plants. Selfing homostyles originate within distylous populations and frequently evolve to become reproductively isolated species. We investigated this process in 10 species of Primula section Obconicolisteri using phylogenomic approaches and inferred how often homostyly originated from distyly and its consequences for population genetic diversity and floral trait evolution. We estimated phylogenetic relationships and reconstructed character evolution using the whole plastome comprised of 76 protein-coding genes. To investigate mating patterns and genetic diversity we screened 15 microsatellite loci in 40 populations. We compared floral traits among distylous and homostylous populations to determine how phenotypically differentiated homostyles were from their distylous ancestors. Section Obconicolisteri was monophyletic and we estimated multiple independent transitions from distyly to homostyly. High selfing rates characterised homostylous populations and this was associated with reduced genetic diversity. Flower size and pollen production were reduced in homostylous populations, but pollen size was significantly larger in some homostyles than in distylous morphs. Repeated transitions to selfing in section Obconicolisteri are likely to have been fostered by the complex montane environments that species occupy. Unsatisfactory pollinator service is likely to have promoted reproductive assurance in homostyles leading to subsequent population divergence through isolation.

The complete chloroplast genome of an endangered endemic herb species in China, Primula filchnerae (Primulaceae).

Primula filchnerae (Primulaceae) is an endangered endemic herb species in China. In this study, we characterized the complete chloroplast genome of P. filchnerae based on next generation sequencing (NGS). The chloroplast genome of P. filchnerae was 151,547 bp in size, containing a large single-copy (LSC) region of 82, 662 bp and a small single-copy (SSC) region of 17,749 bp. These two regions were separated by a pair of inverted repeat regions (IRs), each of 25,568 bp. A total of 130 functional genes were encoded, consisted of 86 protein-coding genes, 36 tRNA genes, and eight rRNA genes.

Aerobic Exercise Training Alleviates Renal Injury by Interfering with Mitochondrial Function in Type-1 Diabetic Mice.

BACKGROUND Diabetic nephropathy was one of the most serious and harmful diabetic complications, characterized by progressive loss of renal function and renal fibrosis. Aerobic exercise training is an important non-pharmacologic method to prevent and treat diabetes mellitus and diabetic complications. MATERIAL AND METHODS Intraperitoneal (i.p.) injection of streptozocin (STZ) was used to construct a type 1 diabetic mouse model. Renal function and mitochondrial function were measured by urinary protein level, Masson staining and ATP, superoxide production, and membrane potential, respectively. The purpose of the research was to explore the effect of aerobic exercise training on renal and renal mitochondrial function, as well as the expression of Sirt1and PGC1α in type-1 diabetic mice. RESULTS Sedentary diabetic mice exhibited increased urinary protein level, blood glucose, and collagen deposition in renal tissues compared with sedentary control mice, which were significantly mitigated by aerobic exercise training. Diabetic mice displayed renal tissue mitochondrial dysfunction (decreased mitochondrial ATP production and membrane potential), as well as increased mitochondrial superoxide production, which were reversed by aerobic exercise. By using Western blot analysis, we identified the decreased expression of Sirt1 and PGC1α in the renal tissue of diabetic mice, which were partly reversed by aerobic exercise training. Data showed that silencing of Sirt1 abrogated the beneficial effect of aerobic exercise training against diabetes-induced mitochondrial abnormalities and renal damage in mice. CONCLUSIONS Aerobic exercise training alleviates diabetes-induced renal injury by improving mitochondrial function.

Phylogeographic insights on the evolutionary breakdown of heterostyly.

The breakdown of heterostyly to homostyly is a classic system for the investigation of evolutionary transitions from outcrossing to selfing. Loss of sexual polymorphism is characterized by changes to population morph structure and floral morphology. Here, we used molecular phylogeography to investigate the geographical context for the breakdown process in Primula chungensis, a species with distylous and homostylous populations. We genotyped plants from 20 populations throughout the entire range in south-west China using the chloroplast intergenic spacer (trnL-trnF), nuclear internal transcribed spacer (ITS) and 10 nuclear microsatellite loci, and determined the genetic relationships among populations and the variation in floral traits associated with homostyle evolution. The marker data identified two multi-population lineages (Tibet and Sichuan) and one single-population lineage (Yunnan), a pattern consistent with at least two independent origins of homostyly. Evidence from flower and pollen size variation is consistent with the hypothesis that transitions to selfing have arisen by the same genetic mechanism involving recombination and/or mutation at the distyly linkage group. Nevertheless, flowers of homostylous lineages have followed divergent evolutionary trajectories following their origin, resulting in populations with both approach and reverse herkogamy. Our study illustrates a rare example of the near-complete replacement of sexual polymorphism by floral monomorphism in a heterostylous species.