Comparison of 5-ALA-PDT and LEEP of cervical squamous intraepithelial neoplasia (CIN2) with high-risk human papillomavirus infection in childbearing age women: A non-randomized controlled polit study.

BACKGROUND: 5-Aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT) is a possible minimally-invasive treatment for high-grade cervical intraepithelial neoplasia (HSIL). The present study was carried out to assess the effect of 5-ALA-PDT and loop electrosurgical excision procedure (LEEP) in cervical squamous intraepithelial neoplasia (CIN2) combined with high-risk human papillomavirus (HR-HPV) infection. METHODS: In this study, 190 patients with CIN2 and HR-HPV infection were finally included. They were divided into the LEEP Group (n = 116) and PDT Group (n = 74) according to gynecologist's recommendation and patient's willingness. All patients were followed up at 4-6 months and 12 months after treatment, including HPV testing, cytology, and colposcopy examination. RESULTS: (1) 4-6 months after treatment, the pathological regression rate was 97.30 % (72/74) in the PDT group and 98.28 % (114/116) in the LEEP group (P = 0.952). The HPV clearance rate was 81.08 % (60/74) in the PDT group and 80.17 % (93/116)in the LEEP group (P = 0.877). (2) 12 months after treatment, the pathological regression rate was 93.24 % (69/74) in the PDT group and 96.55 % (112/116) in the LEEP group (P = 0.486). The recurrence rate of CIN2 was 4.05 % (3/74) in the PDT group and 1.72 % (2/116) in the LEEP group (P = 0.608). The HPV clearance rate was 90.54 % (67/74) in the PDT group and 89.66 % (104/116)in the LEEP group (P = 0.843). The reinfection rate of HR-HPV was 5.41 % (4/74) in the PDT group and 1.72 % (2/116) in the LEEP group (P = 0.322). (3) The adverse reactions in the PDT Group were slightly lower than that in the LEEP Group (P = 0.4956), but the incidence of vaginal bleeding in the PDT group was lower than that in the LEEP group during follow-up. CONCLUSIONS: The effectiveness of 5-ALA-PDT is similar to LEEP for CIN2 with less side effects. Therefore, 5-ALA-PDT, a non-invasive treatment, may be an effective method for CIN2 patients of childbearing age.

Warming enhances the negative effects of shrub removal on phosphorus mineralization potential.

Shrubs have developed various mechanisms for soil phosphorus utilization. Shrub encroachment caused by climate warming alters organic phosphorus mineralization capability by promoting available phosphorus absorption and mediating root exudates. However, few studies have explored how warming regulates the effects of dominant shrubs on soil organic phosphorus mineralization capability. We provide insights into warming, dominant shrub removal, and their interactive effects on the soil organic phosphorus mineralization potential in the Qinghai-Tibetan Plateau. Real-time polymerase chain reaction was used to quantify the soil microbial phosphatase genes (phoC and phoD), which can characterize the soil organic phosphate mineralization potential. We found that warming had no significant effect on the soil organic phosphate-mineralized components (total phosphate, organic phosphate, and available phosphate), genes (phoC and phoD), or enzymes (acid and alkaline phosphatases). Shrub removal negatively influenced the organic phosphate-mineralized components and genes. It significantly decreased soil organic phosphate mineralization gene copy numbers only under warming conditions. Warming increased fungal richness and buffered the effects of shrub removal on bacterial richness and gene copy numbers. However, the change in the microbial community was not the main factor affecting organic phosphate mineralization. We found only phoC copy number had significant correlation to AP. Structural equation modelling revealed that shrub removal and the interaction between warming and shrub removal had a negative direct effect on phoC copy numbers. We concluded that warming increases the negative effect of shrub removal on phosphorus mineralization potential, providing a theoretical basis for shrub encroachment on soil phosphate mineralization under warming conditions.

Comparison of ALA-PDT and CO2 laser treatment of low-grade vaginal intraepithelial neoplasia with high-risk HPV infection: A non-randomized controlled pilot study.

OBJECTIVE: To evaluate the efficacy and safety of 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) and CO2 laser therapy of low-grade vaginal intraepithelial neoplasia (VAIN1) combined with high-risk human papillomavirus (hr-HPV) infection. METHODS: A total of 163 patients with VAIN1 and hr-HPV infection were divided into PDT Group (n = 83) and CO2 laser Group (n = 80). The PDT Group received six times of ALA-PDT treatments and the CO2 laser Group received once CO2 laser treatment. HPV types, cytology, colposcopy, and pathological examinations were carried out before and after treatment. The differences in HPV clearance rate, VAIN1 regression rate, and adverse reactions between the two groups were analyzed during 6-month follow-up. RESULTS: The overall HPV clearance rate of the PDT Group was significantly higher than that of the CO2 laser Group (65.06% vs 38.75%, P = 0.0008) although similar result was obtained for 16/18-related HPV infection patients (54.55% vs 43.48%, P = 0.4578). The VAIN1 regression rate of the PDT Group was significantly higher than that of the CO2 laser Group (95.18% vs 83.75%, P = 0.0170). In patients ≥ 50 years old, ALA-PDT showed better HPV clearance rate and VAIN1 regression rate than CO2 laser therapy (P < 0.05). The adverse reactions in the PDT Group were significantly lower than that in the CO2 laser Group (P > 0.05). CONCLUSIONS: The efficacy of ALA-PDT appears better than CO2 laser for VAIN1 patients. However, the long-term effect of ALA-PDT for VAIN1 still needs to be explored. As a non-invasive treatment, ALA-PDT is a highly effective therapeutic procedure for VAIN1 with hr-HPV infection.

Dual-specificity phosphatase 26-dificient neurons are susceptible to oxygen-glucose deprivation/reoxygenation-evoked apoptosis and proinflammatory response by affecting the TAK1-medaited JNK/P38 MAPK pathway.

Dual-specificity phosphatase 26 (DUSP26) is linked to a broad range of human disorders as it affects numerous signaling cascades. However, the involvement of DUSP26 in ischemic stroke has not been explored. Here, we investigated DUSP26 as a key mediator of oxygen-glucose deprivation/reoxygenation (OGD/R)-associated neuronal injury, an in vitro model for investigating ischemic stroke. A decline in DUSP26 occurred in neurons suffering from OGD/R. A deficiency in DUSP26 rendered neurons more susceptible to OGD/R by aggravating neuronal apoptosis and inflammation, while the overexpression of DUSP26 blocked OGD/R-evoked neuronal apoptosis and inflammation. Mechanistically, enhanced phosphorylation of transforming growth factor-ß-activated kinase 1 (TAK1), c-Jun N-terminal kinase (JNK) and P38 mitogen-activated protein kinase (MAPK) was evidenced in DUSP26-deficient neurons suffering from OGD/R, whereas the opposite effects were observed in DUSP26-overexpressed neurons. Moreover, the inhibition of TAK1 abolished the DUSP26-deficiency-elicited activation of JNK and P38 MAPK and exhibited anti-OGD/R injury effects in DUSP26-deficiency neurons. Results from these experiments show that DUSP26 is essential for neurons in defending against OGD/R insult, while neuroprotection is achieved by restraining the TAK1-mediated JNK/P38 MAPK pathway. Therefore, DUSP26 may serve as a therapeutic target for the management of ischemic stroke.

Direct and indirect effects of dominant plants on ecosystem multifunctionality.

Biodiversity is essential for the provision of multiple ecosystem functions simultaneously (ecosystem multifunctionality EMF). Yet, it remains unclear whether and how dominant plant species impact EMF. Here, we aimed at disentangling the direct from indirect above- and belowground pathways by which dominant plant species influence EMF. We evaluated the effects of two dominant plant species (Dasiphora fruticosa, and the toxic perennial plant Ligularia virgaurea) with expected positive and negative impacts on the abiotic environment (soil water content and pH), surrounding biological communities (plant and nematode richness, biomass, and abundance in the vicinity), and on the EMF of alpine meadows, respectively. We found that the two dominant plants enhanced EMF, with a positive effect of L. virgaurea on EMF greater than that of D. fruticosa. We also observed that dominant plants impacted on EMF through changes in soil water content and pH (indirect abiotic effects), but not through changes in biodiversity of surrounding plants and nematodes (indirect biotic pathway). Our study suggests that dominant plants may play an important role in promoting EMF, thus expanding the pervasive mass-ratio hypothesis originally framed for individual functions, and could mitigate the negative impacts of vegetation changes on EMF in the alpine meadows.

Practical Model for Residual/Recurrent Cervical Intraepithelial Lesions in Patients with Negative Margins after Cold-Knife Conization.

Objective: This study aimed to identify reliable risk factors for residual/recurrent cervical intraepithelial lesions in patients with negative margins after cold-knife conization. Methods: A total of 2352 women with HSILs (high-grade squamous intraepithelial lesions) with negative margins who underwent cold-knife conization between January 2014 and December 2020 were included; in total, 1411 women were assigned to the development cohort, and 941 women were assigned to the validation cohort. Multivariate logistic regression was used to build four predictive models based on the different combinations of follow-up data (Model A: preoperative factors; Model B: first-follow-up data; Model C: second-follow-up data; Model D: data from both follow-ups). The accuracy, sensitivity, specificity, false-positive rate (FPR), false-negative rate (FNR), and area under the receiver operating characteristic curve (AUC) were evaluated on the validation cohort. The predictive power of risk factors was further validated using six machine learning algorithms. Results: Model D demonstrated the highest AUC of 0.91 (95% CI, 0.87 to 0.96) in the validation cohort, whereas Models A, B, and C achieved AUCs of 0.69 (95% CI, 0.59 to 0.78), 0.88 (95% CI, 0.80 to 0.95), and 0.89 (95% CI, 0.81 to 0.97) respectively. The six machine learning methods achieved consistent results. Kaplan-Meier (KM) survival curves demonstrated that our models could effectively stratify patients with all models (p < 0.05 for all models). Conclusion: Our model, which is based on preoperative and follow-up factors, can serve as a complementary screening procedure for the early detection or prediction of recurrence after cold-knife conization in HSIL patients.

Genome-Wide Analysis of the WOX Transcription Factor Genes in Dendrobium catenatum Lindl.

The WUSCHEL-related homeobox (WOX) proteins are a class of transcription factors exclusive to plants. They can promote cell division or inhibit stem cell differentiation to regulate plant growth and development. However, the WOX transcription factor genes in the monocotyledon Dendrobium catenatum Lindl. remain relatively uncharacterized. Specifically, the effects of phytohormones on their expression levels are unclear. In this study, we identified and analyzed 10 candidate DcaWOX transcription factor genes in D. catenatum. The DcaWOX family was divided into the modern/WUS, intermediate, and ancient clades. The subcellular localization analysis detected DcaWOX-GFP fusion proteins in the tobacco epidermal leaf cell nucleus. In DcaWOX, members of the WUS clade with the WUS-box motif can significantly activate the expression of TPL in vivo, while members of the intermediate and ancient clades cannot. The expression of the DcaWOX genes varied among the examined tissues. Moreover, the DcaWOX expression patterns were differentially affected by the phytohormone treatments, with differences detected even between homologs of the same gene. Furthermore, the gene expression patterns were consistent with the predicted cis-acting elements in the promoters. The above results suggest that DcaWOX may have an important role in its growth and development and resistance to stress. The results of this comprehensive investigation of the DcaWOX gene family provide the basis for future studies on the roles of WOX genes in D. catenatum.

Effects of Self-Made Prescription Compound Rhodiola on the Ultrastructure of Podocytes in Rats with Type 2 Diabetic Nephropathy.

Background: We attempt to discuss the function of self-made prescription compound Rhodiola rosea on the kidneys of rats with type 2 diabetic nephropathy (DN), through studying the effects of self-made rescription compound Rhodiola rosea on the ultrastructure of podocytes in rats with DN. Methods: DN rat model was established by streptozotocin. The successfully modeled rats were divided into 4 groups, DN group, compound Rhodiola low-dose group, medium-dose group, and high-dose group. Compound Rhodiola low-dose group, medium-dose group, and high-dose group were administered for 8 weeks, and the DN group and the blank control group were administered with normal saline. The podocyte count, podocyte width, podocyte fusion rate, and average thickness of glomerular basement membrane were compared in each group, and the ultrastructural changes in podocytes were observed by transmission electron microscope. Results: Compared with the normal control group, the number of podocytes in the DN group remarkably reduced, but the width level of podocyte, the fusion rate of podocyte, and the average thickness of basilar membrane remarkably increased (P < 0.05). Compared with the DN group, the number of podocytes in the high-, medium-, and low-dose groups increased remarkably, but the width level of podocyte, the fusion rate of podocyte, and the average thickness of basilar membrane decreased remarkably (P < 0.05). Compared with the low-dose group, the number of podocytes in the high-dose group and the medium-dose group increased remarkably, but the width of podocyte, the fusion rate of podocyte, as well as the average thickness of basilar membrane remarkably reduced (P < 0.05). Various indicators of high- and medium-dose groups had no statistical difference (P > 0.05). Conclusions: Self-made prescription compound Rhodiola rosea can promote the recovery of podocyte in DN rat and protect their kidney.

Multiple lines of evidence supports the two varieties of Halenia elliptica (Gentianaceae) as two species.

Delimiting species requires multiple sources of evidence. Here, we delimited two varieties of Halenia elliptica (Gentianaceae) using several lines of evidence, including morphological traits and mating system in a sympatric population, phylogenetic relationships based on nrITS and cpDNA (rpl16) data, and complete chloroplast genome sequences. Comparative analysis of 21 morphological traits clearly separates the two varieties of H. elliptica. Examination of the flowering process and pollination treatments indicate that H. elliptica var. grandiflora produces seeds via outcrossing, whereas H. elliptica var. elliptica produces seeds via mixed mating. Furthermore, hand-pollinated hybridization of the two varieties produced no seeds. Observations of pollinators showed that when bees began a pollination bout on H. elliptica var. grandiflora they preferred to continue pollinating this variety; however, when they began a pollination bout on H. elliptica var. elliptica, they showed no preference for either variety. Phylogenetic analysis confirmed the monophyly of H. elliptica, which was further divided into two monophyletic clades corresponding to the two varieties. A large number of variants from the chloroplast genomes reflected remarkable genetic dissimilarities between the two varieties of H. elliptica. We recommend that the two varieties of H. elliptica should be revised as two species (H. elliptica and H. grandiflora). Our findings indicate that H. elliptica varieties may have split into two separate species due to a shift in mating system, changes in flowering phenology and/or post-pollination reproductive isolation.

PPARα alleviates iron overload-induced ferroptosis in mouse liver.

Ferroptosis is an iron-dependent form of non-apoptotic cell death implicated in liver, brain, kidney, and heart pathology. How ferroptosis is regulated remains poorly understood. Here, we show that PPARα suppresses ferroptosis by promoting the expression of glutathione peroxidase 4 (Gpx4) and by inhibiting the expression of the plasma iron carrier TRF. PPARα directly induces Gpx4 expression by binding to a PPRE element within intron 3. PPARα knockout mice develop more severe iron accumulation and ferroptosis in the liver when fed a high-iron diet than wild-type mice. Ferrous iron (Fe2+ ) triggers ferroptosis via Fenton reactions and ROS accumulation. We further find that a rhodamine-based "turn-on" fluorescent probe(probe1) is suitable for the in vivo detection of Fe2+ . Probe1 displays high selectivity towards Fe2+ , and exhibits a stable response for Fe2+ with a concentration of 20 µM in tissue. Our data thus show that PPARα activation alleviates iron overload-induced ferroptosis in mouse livers through Gpx4 and TRF, suggesting that PPARα may be a promising therapeutic target for drug discovery in ferroptosis-related tissue injuries. Moreover, we identified a fluorescent probe that specifically labels ferrous ions and can be used to monitor Fe2+ in vivo.

Genome-Wide Analysis of LBD Transcription Factor Genes in Dendrobiumcatenatum.

The LATERAL ORGAN BOUNDARIES DOMAIN (LBD) gene family comprises plant-specific transcription factors that control cell proliferation and differentiation during growth and development in many plant species. However, to date, no studies of the LBD gene family in Dendrobium catenatum have been reported. In this study, a genome-wide analysis of LBD genes was performed in D. catenatum and 24 LBD genes were identified. The genes were classified into two classes (I and II) based on phylogenetic relationships and motif structure. Subcellular localization analysis for DcaLBD6 and DcaLBD18 from class I and DcaLBD37 and DcaLBD41 from class II revealed that the proteins were localized in the nucleus. Transient expression analysis of DcaLBD6, DcaLBD18, DcaLBD37, and DcaLBD41 indicated that class I and class II members have opposite roles in regulating VASCULAR-RELATED NAC-DOMAIN 7 (VND7) expression. DcaLBD genes showed diverse expression patterns in response to different phytohormone treatments. Heat maps revealed diverse patterns of DcaLBD gene expression in different organs. These results lay the foundation for further detailed studies of the LBD gene family in D. catenatum.

Metabolic nuclear receptors coordinate energy metabolism to regulate Sox9+ hepatocyte fate.

Recent research has indicated the adult liver Sox9+ cells located in the portal triads contribute to the physiological maintenance of liver mass and injury repair. However, the physiology and pathology regulation mechanisms of adult liver Sox9+ cells remain unknown. Here, PPARα and FXR bound to the shared site in Sox9 promoter with opposite transcriptional outputs. PPARα activation enhanced the fatty acid ß-oxidation, oxidative phosphorylation (OXPHOS), and adenosine triphosphate (ATP) production, thus promoting proliferation and differentiation of Sox9+ hepatocytes along periportal (PP)-perivenous (PV) axis. However, FXR activation increased glycolysis but decreased OXPHOS and ATP production, therefore preventing proliferation of Sox9+ hepatocytes along PP-PV axis by promoting Sox9+ hepatocyte self-renewal. Our research indicates that metabolic nuclear receptors play critical roles in liver progenitor Sox9+ hepatocyte homeostasis to initiate or terminate liver injury-induced cell proliferation and differentiation, suggesting that PPARα and FXR are potential therapeutic targets for modulating liver regeneration.

Computer-aided diagnostic system based on deep learning for classifying colposcopy images.

BACKGROUND: Colposcopy is widely used to detect cervical cancer, but developing countries lack the experienced colposcopists necessary for accurate diagnosis. Artificial intelligence (AI) is being widely used in computer-aided diagnosis (CAD) systems. In this study, we developed and validated a CAD model based on deep learning to classify cervical lesions on colposcopy images. METHODS: Patient data, including clinical information, colposcopy images, and pathological results, were collected from Qilu Hospital. The study included 15,276 images from 7,530 patients. We performed two tasks in this study: normal cervix (NC) vs. low grade squamous intraepithelial lesion or worse (LSIL+) and high-grade squamous intraepithelial lesion (HSIL)- vs. HSIL+. The residual neural network (ResNet) probability was calculated for each patient to reflect the probability of lesions through a ResNet model. Next, a combination model was constructed by incorporating the ResNet probability and clinical features. We divided the dataset into a training set, validation set, and testing set at a ratio of 7:1:2. Finally, we randomly selected 300 patients from the testing set and compared the results with the diagnosis of a senior colposcopist and a junior colposcopist. RESULTS: The model that combines ResNet and clinical features performs better than ResNet alone. In the classification of NC and LSIL+, the area under the receiver operating characteristic curve (AUC), accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 0.953, 0.886, 0.932, 0.846, 0.838, and 0.936, respectively. In the classification of HSIL- and HSIL+, the AUC, accuracy, sensitivity, specificity, PPV, and NPV were 0.900, 0.807, 0.823, 0.800, 0.618, and 0.920, respectively. In the two classification tasks, the diagnostic performance of the model was determined to be comparable to that of the senior colposcopist and exhibited a stronger diagnostic performance than the junior colposcopist. CONCLUSIONS: The CAD system for cervical lesion diagnosis based on deep learning performs well in the classification of cervical lesions and can provide an objective diagnostic basis for colposcopists.

Direct and indirect facilitation affect community productivity through changes in functional diversity in an alpine system.

BACKGROUND AND AIMS: Facilitation is an important ecological process for plant community structure and functional composition. Although direct facilitation has accrued most of the evidence so far, indirect facilitation is ubiquitous in nature and it has an enormous potential to explain community structuring. In this study, we assess the effect of direct and indirect facilitation on community productivity via taxonomic and functional diversity. METHODS: In an alpine community on the Tibetan Plateau, we manipulated the presence of the shrub Dasiphora fruticosa and graminoids in a fenced meadow and a grazed meadow to quantify the effects of direct and indirect facilitation. We measured four plant traits: height, lateral spread, specific leaf area (SLA) and leaf dry matter content (LDMC) of forbs; calculated two metrics of functional diversity [range of trait and community-weighted mean (CWM) of trait]; and assessed the responses of functional diversity to shrub facilitation. We used structural equation modelling to explore how shrubs directly and indirectly drove community productivity via taxonomic diversity and functional diversity. KEY RESULTS: We found stronger effects from herbivore-mediated indirect facilitation than direct facilitation on productivity and taxonomic diversity, regardless of the presence of graminoids. For functional diversity, the range and CWM of height and SLA, rather than lateral spread and LDMC, generally increased due to direct and indirect facilitation. Moreover, we found that the range of traits played a primary role over taxonomic diversity and CWM of traits in terms of shrub effects on community productivity. CONCLUSIONS: Our study reveals that the mechanism of shrub direct and indirect facilitation of community productivity in this alpine community is expanding the realized niche (i.e. expanding range of traits). Our findings indicate that facilitators might increase trait dispersion in the local community, which could alleviate the effect of environmental filters on trait values in harsh environments, thereby contributing to ecosystem functioning.

Development of Microsatellite Markers for a Dioecious Herpetospermum pedunculosum (Cucurbitaceae).

Understanding the evolution of flower diversity is a central topic in plant evolutionary ecology, and natural selection on floral traits via male fitness could be estimated quantitatively using microsatellites. Here, based on RNA sequencing, we developed simple sequence repeat primers and verified polymorphisms in 2 wild populations of Herpetospermum pedunculosum (Cucurbitaceae), a dioecious annual plants native to the Himalaya Mountains. A total of 131 paired primers were designed; 15 paired primers were found to be polymorphic, with the expected heterozygosity varying between 0.280 and 0.767. We also identified 58 genotypes in 20 plants from the 2 populations. Conclusively, these primers could be effective in examining male fitness and population genetic structure of H pedunculosum in future studies.

IL-6 expression promoted by Poly(I:C) in cervical cancer cells regulates cytokine expression and recruitment of macrophages.

Poly(I:C) is a promising adjuvant for cancer treatment vaccines to enhance the host anti-tumour immune response. However, the roles of poly(I:C) in the cervical cancer microenvironment and local immune reactions are not well understood. In this study, we investigated the roles of poly(I:C) in the cervical cancer. We analysed the cytokine transcription and secretion of cervical cancer cell lines and THP-1-derived macrophages after poly(I:C) treatment, respectively. These results revealed that IL-6 was significantly up-regulated, and this up-regulation was partly dose dependent. poly(I:C)-stimulated supernatant of cervical cancer cells promoted M1-type cytokine IL-1ß and IL-6 expression of THP-1-derived macrophages, but inhibited the expression of M2-type cytokine, IL-10 and CCL22. The recruitment of THP-1-derived macrophages by poly(I:C)-stimulated cervical cancer cell supernatant was also enhanced. Inhibition of IL-6 expression in cervical cancer cells by siRNA transfection almost completely reversed the effects of poly(I:C) treatment. Finally, we found that phosphorylation of the NF-κB signalling pathway in cervical cancer cells occurred quickly after poly(I:C) treatment. Moreover, the NF-κB signalling pathway inhibitor PDTC significantly inhibited poly(I:C)-induced IL-6 expression. Taken together, these results suggest that poly(I:C) might regulate the effects of cervical cancer cells on tumour-infiltrated macrophages, and subsequently promote a pro-inflammatory tumour microenvironment.

Transcriptome Analysis and Microsatellite Markers Development of a Traditional Chinese Medicinal Herb Halenia elliptica D. Don (Gentianaceae).

Halenia elliptica is a popular Chinese medicinal herb that is used to treat jaundice disease and virus hepatitis, and its wild populations have been reduced significantly due to overharvesting recently. However, effective conservation could not be implemented because of the lack of genomic information and genetic markers. In this study, a de novo transcriptome of H elliptica was sequenced using the NGS Illumina, and 132 695 unigenes with the length >200 bp (base pairs) were obtained. Among them, a total of 32 109 unigenes were scanned to develop simple sequence repeats (SSRs). Based on NCBI (National Center for Biotechnology Information) nonredundant database (Nr), these SSR sequences were annotated and assigned into gene ontology categories. In addition, we designed 126 pairs of SSR primers for polymerase chain reaction amplification, of which 12 pairs were identified to be polymorphic among 40 individuals from 8 populations. We then used the 12 polymorphic SSRs to construct a UPGMA dendrogram of the 40 individuals. In addition, a significant correlation between the genetic relationship and the geographic distance was found, suggesting a phylogeographic structure in H elliptica. Moreover, 2 of these SSRs were also successfully amplified in a related species Veratrilla baillonii, suggesting their cross-species transferability. Generally, the SSR markers with high polymorphisms identified in this study provide valuable genetic resources and represent an initial step for exploring the genetic diversity and population histories of H elliptica and its related species.

Equipped for Migrations Across High Latitude Regions? Reduced Spur Length and Outcrossing Rate in a Biennial Halenia elliptica (Gentianaceae) With Mixed Mating System Along a Latitude Gradient.

Halenia (Gentianaceae) originated from the mountain regions of East Asia, and diversified in America following long migrations via Beringia. While Halenia elliptica, one species of the genus in China, migrated toward high latitudes in China. Spur length of H. elliptica is highly variable. We examined the relationship between spur length and mating pattern along a latitude gradient. Field experiments were performed in two populations of H. elliptica, and we found that this species could produce seeds via both autonomous selfing and the aid of pollinators, suggesting a mixed mating system. In seven populations of H. elliptica along a latitudinal gradient, we found a trend of decrease in spur length with the increase of latitude. Based on molecular data from 11 microsatellite loci, we found that multilocus outcrossing rate decreased with the increase of latitude while the estimated inbreeding depression increased significantly, indicating that a high degree of inbreeding depression might have prevented evolution toward complete selfing in the high latitude populations with short spur length, and thus maintained mixed mating system of H. elliptica. Our results suggest that the mixed mating system of this species might be helpful in overcoming pollinator scarcity in newly colonized populations toward high latitudes after its origination in the mountain regions of China, and the decrease of spur length in the high latitude populations could result from reduced resource allocation to pollinator associated traits.

Differentiation in drought tolerance mirrors the geographic distributions of alpine plants on the Qinghai-Tibet Plateau and adjacent highlands.

Climatic tolerance, especially drought tolerance, is one of the major factors shaping the geographic distributions of plant species. Thus, the general decline in rainfall from the Himalaya-Hengduan Mountains (HHM) to the inner Qinghai-Tibet Plateau (QTP) might account for the significant differences in species distributions and richness between the two regions. To test this hypothesis, we conducted a water stress experiment using four Anisodus species (A. tanguticus, A. luridus, A. carniolicoides and A. acutangulus), which were treated with different levels of water stress in a glasshouse, and examined their differences in physiological responses. The results suggest that A. tanguticus, which inhabits the inner QTP, generally has higher fitness under severe water stress than the other species based on its high root:shoot ratio, long-term water use efficiency and photosynthetic rate, indicating that it possesses a genetically based drought tolerance mechanism. Our results suggest that plant species inhabiting the inner QTP may be more drought tolerant than those inhabiting the HHM regions. This provides a new example supporting the hypothesis that climatic tolerance plays a major role in shaping plant distributions on the QTP and its adjacent highlands and presents new insights into the patterns of geographic distribution and diversity of the plants inhabiting these areas.

The complete chloroplast genome of Sinopodophyllum hexandrum Ying (Berberidaceae).

The complete nucleotide sequence of the Sinopodophyllum hexandrum Ying chloroplast genome (cpDNA) was determined based on next-generation sequencing technologies in this study. The genome was 157 203 bp in length, containing a pair of inverted repeat (IRa and IRb) regions of 25 960 bp, which were separated by a large single-copy (LSC) region of 87 065 bp and a small single-copy (SSC) region of 18 218 bp, respectively. The cpDNA contained 148 genes, including 96 protein-coding genes, 8 ribosomal RNA genes, and 44 tRNA genes. In these genes, eight harbored a single intron, and two (ycf3 and clpP) contained a couple of introns. The cpDNA AT content of S. hexandrum cpDNA is 61.5%.