NUCB2 inhibition antagonizes osteosarcoma progression and promotes anti-tumor immunity through inactivating NUCKS1/CXCL8 axis.

The oncogenic properties of Nucleobindin2 (NUCB2) have been observed in various cancer types. Nevertheless, the precise understanding of the biological functions and regulatory mechanisms of NUCB2 in osteosarcoma remains limited. This investigation reported that NUCB2 was significantly increased upon glucose deprivation-induced metabolic stress. Elevated NUCB2 suppressed glucose deprivation-induced cell death and reactive oxygen species (ROS) increase. Depletion of NUCB2 resulted in a reduction in osteosarcoma cell proliferation as well as metastatic potential in vitro and in vivo. Mechanically, NUCB2 ablation suppressed C-X-C Motif Chemokine Ligand 8 (CXCL8) expression which then reduced programmed cell death 1 ligand 1 (PD-L1) expression and stimulated anti-tumor immunity mediated through cytotoxic T cells. Importantly, a combination of NUCB2 depletion with anti-PD-L1 treatment improved anti-tumor T-cell immunity in vivo. Moreover, we further demonstrated that NUCB2 interacted with NUCKS1 to inhibit its degradation, which is responsible for the transcriptional regulation of CXCL8 expression. Altogether, the outcome emphasizes the function of NUCB2 in osteosarcoma and indicates that NUCB2 elevates osteosarcoma progression and immunosuppressive microenvironment through the NUCKS1/CXCL8 pathway.

HOXB9 promotes osteosarcoma cell survival and malignancy under glucose starvation via upregulating SPP1 expression.

Homeobox B9 (HOXB9) has been shown to play a critical role in several tumors. However, the precise biological mechanisms and functions of HOXB9 in osteosarcoma remain largely unknown. In this study, we found that HOXB9 was increased upon glucose starvation. Elevated HOXB9 suppressed osteosarcoma cell death and supported cell growth and migration under glucose starvation. Further mechanistic studies demonstrated that HOXB9 directly bound to the promoter of secreted phosphoprotein 1 (SPP1) and transcriptionally upregulated SPP1 expression which then led cell death decrease and cell growth increase under glucose deprivation environment. Clinically, HOXB9 was significantly upregulated in osteosarcoma compared with normal tissues and increase of HOXB9 expression was positively associated with the elevation of SPP1 in osteosarcoma. Overall, our study illustrates that HOXB9 contributes to malignancy in osteosarcoma and inhibits cell death through transcriptional upregulating SPP1 under glucose starvation.

Dietary therapy of the herbal porridge improves the symptoms of functional dyspepsia: A randomized, double-blind, placebo-controlled, clinical trial.

This study (ISRCTN17174559) aimed to explore the efficacy and safety of a kind of herbal porridge (Hou Gu Mi Xi) on the clinical symptoms of functional dyspepsia (FD). This was a single-center, single-dose, prospective, double-blind, randomized controlled trial involving 64 participants with FD (35 cases and 29 controls) for 2 months of intervention and 1 month of follow-up. The 7-point Global Overall Symptom Scale (GOSS), 36-Item Short Form Survey (SF-36), and other indicators were assessed at baseline (day 0), at days 15, 30, and 60 of treatment, and at follow-up 1 month after the end of the intervention. Many participants with FD achieved remission of their epigastric symptoms at follow-up on the 90th day after treatment with herbal porridge compared to the placebo group (45.71% vs. 20.69%, p = .036). Furthermore, herbal porridge appeared to be effective in improving the quality of life of participants with FD, which was reflected in the rising SF-36 scores for physical role, bodily pain, emotional role, and mental health. Although adverse events were reported, there was no overall difference in the number of adverse events between the two groups (p = .578). Herbal porridge is another effective and safe method for improving the symptoms and quality of life in patients with FD.

BMS-794833 reduces anlotinib resistance in osteosarcoma by targeting the VEGFR/Ras/CDK2 pathway.

Background: Osteosarcoma, a tumor that originates from bone cells, has a poor prognosis and a high degree of malignancy. Anlotinib, a small-molecule multi-target tyrosine kinase inhibitor (TKI), is the first-line drug in treating osteosarcoma, especially in late-stage osteosarcoma. However, patients often develop resistance after using anlotinib for a certain period, which poses a challenge to its further clinical application. Recently, several TKIs, for instance regorafenib and cabozantinib, have showed clinical interest in treating osteosarcoma and target both vascular endothelial growth factor receptor (VEGFR) and mesenchymal epithelial transition factor (c-MET). Therefore, the identification of new TKI warrants further investigation. Methods: We performed CCK8 aasays to confirm that BMS-794833 sensitization osteosarcoma cells to anlotinib. Bioinformatics analysis and rescue experiments showed that the reduce of resistance were dependent on the VEGFR/Ras/CDK2 pathway. Cell line based xenograft model were used to demonstrate that BMS-794833 and anlotinib could synergistically treat OS. Results: Here, we found that BMS-794833 reduced anlotinib resistance in osteosarcoma by targeting the VEGFR/Ras/CDK2 pathway. CCK8 assay showed that BMS-794833 significantly improved the resistance of osteosarcoma cells to anlotinib. The results of rescue experiments showed that the regulatory effects of BMS-794833 on the proliferation and drug resistance of osteosarcoma cells were dependent on the VEGFR/Ras/CDK2 pathway. In addition, BMS-794833 affected the resistance of osteosarcoma cells to anlotinib through epithelial-mesenchymal transition (EMT) and apoptosis pathways. More importantly, BMS-794833 and anlotinib exerted synergistic therapeutic effects against osteosarcoma in vivo. Conclusion: Altogether, this study reveals a new (VEGFR)-targeting drug that can be combined with anlotinib for the treatment of osteosarcoma, which provides an important theoretical basis for overcoming anlotinib resistance.

Association between sugar-free beverage intake and childhood obesity among Chinese children and adolescents.

BACKGROUND: The relationship between sugar-free beverage (SFB) intake and childhood obesity among Chinese children is unknown. OBJECTIVES: To describe the status of SFB consumption among children and adolescents in China and assess the association between SFB intake and different types of obesity. METHODS: The study was based on the baseline data of an ongoing cohort project named Evaluation and Monitoring on School-based Nutrition and Growth in Shenzhen (EMSNGS). Food frequency questionnaires were used to collect information on SFB consumption in 3227 students aged 9-17. Physical and clinical examinations were conducted by trained investigators and clinicians. Multivariable binary logistic regression models were performed to assess the association between SFB intake and general obesity, overweight/obesity, abdominal obesity, metabolically unhealthy overweight (MUOW)/metabolically unhealthy obesity (MUO). RESULTS: The median age of the participants was 13.28 years. Among the participants, 55.2% were boys, and 66.1% were adolescents. The median SFB consumption was 16.67 mL/d. After adjusting for potential confounding factors, each 100 mL increase in daily SFB intake was associated with an increased risk of overweight/obesity (OR = 1.14; 95%CI: 1.06-1.23), abdominal obesity (OR = 1.12; 95%CI: 1.03-1.23), and MUOW/MUO (OR = 1.12; 95%CI: 1.02-1.21), respectively. Stratified analyses showed that family income may have an impact on the association between SFB intake and overweight/obesity (P for interaction = 0.021) and abdominal obesity (P for interaction = 0.031). CONCLUSION: SFB intake was positively associated with childhood obesity in Chinese children, particularly among individuals with high-income families.

Deep eutectic solvent-based ultrasound-assisted extraction in soil samples preparation and elemental determination by ICP-OES.

A fast, simple and green method was established for the extraction of Al, Be, Ca, Co, Cr, Cu, Mn, P, Pb, V and Zn from soil samples using ultrasound-assisted deep eutectic solvents (DESs). DESs based on choline chloride, carboxylic acids and polyols were investigated. It was established that the solvent synthesized from choline chloride and oxalic acid provided the highest extraction recovery (85-104%). Inductively coupled plasma-optical emission spectrometry (ICP-OES) was employed to determine the target analytes in the extracts. The parameters that affect the extraction of target analytes was optimized using standard reference samples. Target analytes could be effectively isolated from soil samples using 0.5 g of DES, ultrasound for 40 min, and heating at 80 â„ƒ. The content of the target analytes determined by this method showed no significant difference from the certified values of 24 reference samples. The proposed method was applied to quantify target analytes in real soil samples. Compared to the traditional acid digestion method, this method showed no significant difference in precision and accuracy, with a confidence level of 95%. The proposed method was found to be simple, accurate and environmentally friendly.

[Comparison of posterolateral approach and combined approach in treatment of Mason type 2B posterior malleolar fracture].

Objective: To compare the effectiveness between the posterolateral approach and the posterolateral combined posteromedial approaches in the treatment of Mason type 2B posterior malleolar fracture. Methods: A retrospective analysis was performed on the clinical data of 79 patients with posterior ankle fracture who met the selection criteria between January 2015 and January 2022. There were 62 cases of Mason 2B Pilon subtype and 17 cases of avulsion subtype. Among Mason 2B Pilon subtype patients, 35 were treated with posterolateral approach (group A), 27 patients were treated with combined approach (group B). There was no significant difference in gender, age, injured side, cause of injury, time from injury to operation, preoperative hospital stay, preoperative visualanalogue scale (VAS) score, and intraoperative internal fixation between the two groups ( P>0.05). All patients with Mason 2B avulsion subtype were treated by posterolateral approach, including 7 males and 10 females, aged from 25 to 68 years, with an average of 46.1 years. The operation time, intraoperative blood loss, postoperative hospital stay, and complications were recorded. The reduction quality was evaluated by Ovadia deals radiographic score, and the ankle function and pain were evaluated by VAS score, American Orthopaedic Foot and Ankle Society (AOFAS) score, and ankle range of motion. Results: Mason 2B Pilon subtype: There was no significant difference in operation time, intraoperative blood loss, postoperative hospital stay, and follow-up time between the two groups ( P>0.05). The radiological evaluation of Ovadia deals in group A was significantly worse than that in group B ( P<0.05). The VAS score in the two groups significantly improved at each time point after operation, and the VAS score and AOFAS score further improved with the extension of time after operation, and the differences were significant ( P<0.05). Except that the AOFAS score of group A was significantly lower than that of group B at last follow-up ( P<0.05), there was no significant difference in VAS score and AOFAS score between the two groups at other time points ( P>0.05). At last follow-up, the ankle range of motion in group A was significantly less than that in group B ( P<0.05). There was no significant difference in the incidence of sural nerve injury, deep tissue infection, limitation of toe movement, and traumatic ankle arthritis between the two groups ( P>0.05). Mason 2B avulsion subtype: The operation time was (119.47±20.61) minutes and the intraoperative blood loss was 50 (35, 55) mL. Seventeen patients were followed up 13-25 months, with an average of 18 months. The Ovadia deals score was excellent in 10 cases, good in 6 cases, and poor in 1 case at 1 week after operation, and the excellent and good rate was 94.1%. All fractures healed in 8-18 weeks with an average of 12.35 weeks. There were 1 case of sural nerve injury and 3 cases of traumatic ankle arthritis after operation. No deep tissue infection or limitation of toe movement occurred. The VAS score decreased significantly and AOFAS score increased significantly with time, and the differences were significant between different time points before and after operation ( P<0.05). The ankle range of motion at last follow-up was (56.71±2.47)°. Conclusion: Compared with the posterolateral approach, the combined approach is a better choice for the treatment of Mason 2B Pilon subtype. If the posteromedial bone block does not affect the reduction of the medial malleolus, the posterolateral approach can achieve good effectiveness for Mason 2B avulsion subtype.

Retrieving soil moisture using cosmic-ray neutron technology based on COSMIC model in the desert-oasis region.

Cosmic-ray neutron technology could estimate average soil moisture on scale of hectometers by monitoring the neutron intensity near the ground, which has been successfully applied in forest, grassland, farmland, and other ecosystems. To verify the reliability of Cosmic-ray Soil Moisture Interaction Code (COSMIC) model for retrieving mesoscale soil moisture in arid regions, we carried out soil moisture observation experiment by using the cosmic-ray neutron rover in the desert-oasis region of the middle reaches of Heihe River. The results showed that the fast neutron intensity in the desert-oasis region were 350-715 counts·(30 s)-1, and the calibrated high energy neutron intensity (Ncosmic) were (38.5±2.2) counts·(30 s)-1, which was affected by land surface characteristics. Both COSMIC model (root mean square error=0.019 g·g-1) and N0 equation (root mean square error=0.018 g·g-1) could well assess the mesoscale soil moisture, with the accuracy of soil moisture being higher considering soil lattice water. The average penetration depth was 19 cm in the oasis region and 36 cm in the desert region during the experiment. COSMIC model could be used to retrieve soil moisture by cosmic ray neutron in the desert-oasis regions, which had great potential to realize data assimilation of surface meteorological-hydrological-ecological variables by combining with land surface models.

Determination of major and trace elements in plant samples by inductively coupled plasma optical emission spectrometry with deep eutectic solvent extraction based on choline chloride and carboxylic acids.

A rapid, precise, and environmentally friendly approach utilizing an ultrasound assisted deep eutectic solvent-based extraction method was developed for the extraction of Ca, Fe, K, Mg, Mn, Na, P, S and Zn from plant samples. The investigation was conducted on deep eutectic solvents that are based on choline chloride and carboxylic acids. The determination of target analytes in the extracts was carried out using inductively coupled plasma-optical emission spectrometry (ICP-OES). The DESs based on malic acid were found to exhibit the highest extraction recovery values (95-106%). The parameters affecting the extraction of target analytes were optimized using standard reference materials. The target analytes can be effectively extracted from plant samples using 0.5 g of DESs and ultrasonication for 40 minutes. The determination results of the reference samples indicated that the relative error (RE) was below 15.1%, and the relative standard deviation (RSD) was less than 6.3%, demonstrating excellent accuracy and precision. The proposed method was employed to quantify target analytes in actual plant samples. The accuracy of this method was not significantly different from that of the microwave digestion method. The proposed method has been demonstrated to be a valid approach for the determination of target elements in actual plant samples.

NUCKS1, a LINC00629-upregulated gene, facilitated osteosarcoma progression and metastasis by elevating asparagine synthesis.

Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 (NUCKS1) has been reported to play an oncogenic role in several cancers. However, the biological functions and regulatory mechanism of NUCKS1 in osteosarcoma have not been fully understood. In this study, we reported that NUCKS1 was significantly increased in osteosarcoma. Depletion of NUCKS1 decreased osteosarcoma cell proliferation and metastasis in vivo and in vitro. Overexpression of NUCKS1 accelerated osteosarcoma cell aggressiveness. Mechanistically, NUCKS1 facilitated asparagine (Asn) synthesis by transcriptionally upregulating asparagine synthetase (ASNS) expression and elevating the levels of Asn in osteosarcoma cells, leading to increased cell growth and metastasis. Inhibition of ASNS or reduction of Asn decreased osteosarcoma cell aggressiveness and impaired the promoting effects of NUCKS1 on tumorigenesis and metastasis. Furthermore, we also found that by acting as a sponge for miR-4768-3p, LINC00629 promoted NUCKS1 expression. Collectively, our findings highlight the role of NUCKS1 in regulating asparagine metabolism and reveal that LINC00629 is an important regulator of NUCKS1 that contributes to NUCKS1 upregulation in osteosarcoma.

Impacts of climate change and anthropogenic stressors on runoff variations in major river basins in China since 1950.

Runoff is one of the main components of hydrological cycle and an important index for water resources evaluation, understanding the runoff change and their causes is vital to water resource management. In the study, we analyzed the runoff change and the impacts of climate change and land use alteration on runoff variation based on natural runoff and previous studies in China. The results showed that there was a significant increasing trend in the annual runoff during 1961-2018 (p < 0.05), with change rate of 0.4 mm/a and abrupt point at 1999 across China, climate change dominated the runoff variation with a contribution of 54 %. In previous studies, the runoff of the major basins in China had a downward trend on the whole (-0.99 mm/a) except Continental River Basin (CRB) showed an increasing trend (0.25 mm/a), the abrupt points were mainly concentrated in 1991-2000, and human activity was the leading factor of runoff change with the contribution of 54 % across China. Human activity was the dominant factor of runoff change in Songhua and Liao River Basin (SLRB), Yellow River Basin (YRB), Hai River Basin (HRB) and Pearl River Basin (PRB), the contribution was >56 %, while climate change was the dominant factor of runoff change in Huai River Basin (HuRB), CRB, and Yangtze River Basin (YZRB). Overall, there was a significant correlation between runoff and precipitation, unused land, urban and grassland in China. We concluded that runoff change and the contribution of climate change and human activities varies greatly among different basins. The findings in this work can shed light on the quantitative understanding of runoff changes in national scale and offer a scientific basis for sustainable water management.

Ecohydrological insight: Solar farms facilitate carbon sink enhancement in drylands.

Solar farms are critical to tackling climate change and achieving carbon neutrality. Besides producing renewable energy, a solar farm modifies microclimates and changes water distribution, consequently affecting local carbon sequestration capacity (CSC). Yet, how the CSC of an ecosystem responds to these changes after solar farm construction remains inadequately understood. Herein, the SOFAR model was adopted to reveal the effects of large-scale solar farms (LSFs) on CSC in arid northern China, with a series of numeric experiments along a climate gradient (with precipitation ranging from 70 to 500 mm yr-1). The results show that relative to pristine vegetation background, CSC was non-linearly increased by averages of 3.49-6.68%, 4.43-10.25%, 5.07-9.71% and 5.6% each year after the installation of LSFs in hyper-arid climates (with aridity index or AI = 0.04-0.05), arid climates (AI = 0.14-0.16), semi-arid climates (AI = 0.21-0.3) and semi-humid climates (AI = 0.55), respectively. The increase in available water for plants growing under the drip lines of photovoltaic panels (PVs) in LSFs is confirmed to be the overwhelming factor responsible for CSC enhancement. Although biases remain in the estimation of increased CSC in hyper- and semi-humid regions due to the high variability of climate (e.g., extreme drought events) and serious radiation reduction beneath PVs, it is certain that solar farms facilitate CSC without increasing external land use. These results will deepen our understanding of the feedback between solar farms and ambient environments and be meaningful for vegetation management in solar farms, especially in the context of climate change and carbon neutrality aims.

The limiting effect of genome size on xylem vessel diameter is shifted by environmental pressures in seed plants.

Current and previous studies have extensively studied the physiological and ecological consequences of genome size (GS) on plants because of the limiting effect of GS on cell size. However, it is still obscure whether such limiting effect could be shifted by environmental pressures, or not. Here, we compiled a global dataset comprised of GS, xylem vessel diameter (V dia), xylem hydraulic conductivity (K S), P 50 (xylem water potential at the loss of 50% maximum K S), and climate factors of 251 phylogeny and habitat divergent species from 59 families. The results showed that GS could limit the V dia of the species from the same family sampled in the similar climate conditions. But the expected positive relationship between GS and V dia became uncertain and even negative across different environmental conditions. V dia was strongly positively coordinated with mean annual temperature (MAT), mean annual precipitation (MAP), and potential evapotranspiration (PET). Furthermore, V dia as the anatomic foundation of plant hydraulic performance was strongly positively coordinated with K S and negatively coordinated with -P 50. The strong environmental selection on K S and P 50 explained the concerted regulation of V dia by environmental factors. The findings revealed the combined regulation of GS and environmental pressures on xylem cell size and thus affected plant eco-physiological performance. The shifted cell size limiting effect of GS by environmental factors manifests plants great plasticity under changed environmental conditions.

LINC00629 protects osteosarcoma cell from ER stress-induced apoptosis and facilitates tumour progression by elevating KLF4 stability.

BACKGROUND: Escaping from ER stress-induced apoptosis plays an important role in the progression of many tumours. However, its molecular mechanism in osteosarcoma remains incompletely understood. METHODS: The molecular mechanism was investigated using RNA sequencing, qRT-PCR and Western blot assays. The relationship between LINC00629 and KLF4 was investigated using RNA pulldown and ubiquitylation assays. The transcriptional regulation of laminin subunit alpha 4 (LAMA4) by KLF4 was identified using bioinformatic analysis, a luciferase assay, and a chromatin immunoprecipitation assay. RESULTS: Here, we demonstrated that LINC00629 was increased under ER stress treatment. Elevated LINC00629 inhibited ER stress-induced osteosarcoma cell apoptosis and promoted clonogenicity and migration in vitro and in vivo. Further mechanistic studies indicated that LINC00629 interacted with KLF4 and suppressed its degradation, which led to a KLF4 increase in osteosarcoma. In addition, we also found that KLF4 upregulated LAMA4 expression by directly binding to its promoter and that LINC00629 inhibited ER stress-induced apoptosis and facilitated osteosarcoma cell clonogenicity and metastasis by activating the KLF4-LAMA4 pathway. CONCLUSION: Collectively, our data indicate that LINC00629 is a critical long noncoding RNA (lncRNA) induced by ER stress and plays an oncogenic role in osteosarcoma cell by activating the KLF4-LAMA4 axis.

Leaf water potential-dependent leaflet closure contributes to legume leaves cool down and drought avoidance under diurnal drought stress.

Efficient thermoregulation under diurnal drought stress protects leaves from photosystem damage and water supply-demand imbalance, yet the cool effect and drought avoidance by leaflet closure have not been well understood. We investigated the cool effect and the drought avoidance of leaflet closure in legume species that survived in the semi-arid region facing seasonal and diurnal drought stress. The results showed that leaflet closure effectively cooled down legume leaves through a reduction of projected leaflet area and the cosine of the angle of incidence (cos i). The leaflet closure was strongly dependent on leaf water potential (Ψleaf). In addition, by characterizing the sequence of key leaf drought response traits, we found leaflet closure occurred after stomatal closure and reduced transpiration rate but before hydraulic failure and turgor loss point (Ψtlp). The meta-analysis also showed that the leaflet closure and cos i decreased after the stomatal conductance declined but before midday. These results imply that Ψleaf-dependent leaflet closure as an alternative to transpiration for leaflet cooling down and as a protective drought avoidance strategy assisting sessile legume plants survival under drought stress.

An amphoteric betaine electrolyte additive enabling a stable Zn metal anode for aqueous batteries.

The corrosion and dendritic growth of the Zn anode limit its electrochemical performance in aqueous Zn batteries. Here, we present an amphoteric betaine additive for 5 m ZnCl2 aqueous electrolyte. The carboxyl group on betaine forms hydrogen bonds with water and reduces the water activity. The molecule also experiences preferential adsorption on the Zn surface and separates the interactions between Zn and water. Side reactions at the Zn electrode are thus inhibited. The regulated interface also ensures uniform Zn deposition. As a result, the electrolyte with betaine additive allows reversible Zn plating/stripping for over 1400 h at 0.5 mA cm-2. A capacity retention of 94% is obtained after 3000 cycles for a VO2 cathode.

The Grain-for-Green project offsets warming-induced soil organic carbon loss and increases soil carbon stock in Chinese Loess Plateau.

The dynamics of soil organic carbon (SOC) stock is a vital element affecting the climate, and ecological restoration is potentially an effective measure to mitigate climate change by enhancing vegetation and soil carbon stocks and thereby offsetting greenhouse gas emissions. The Grain-for-Green project (GFGP) implemented in Chinese Loess Plateau (LP) since 1999 is one of the largest ecological restoration projects in the world. However, the contributions of ecological restoration and climate change to ecosystem soil carbon sequestration are still unclear. In this study, we improved a soil carbon decomposition framework by optimizing the initial SOC stock based on full spatial simulation of SOC and incorporating the priming effect to investigate the SOC dynamics across the LP GFGP region from 1982 through 2017. Our results indicated that SOC stock in the GFGP region increased by 20.18 Tg C from 1982 through 2017. Most portion (15.83 Tg C) of the SOC increase was accumulated when the GFGP was initiated, with a SOC sink of 16.12 Tg C owing to revegetation restoration and a carbon loss of 0.29 Tg C due to warming during this period. The relationships between SOC and forest canopy height and investigations on the SOC dynamics after afforestation revealed that the accumulation rate of SOC could be as high as 24.68 g C m-2 yr-1 during the 70 years following afforestation, and that SOC could decline thereafter (-8.89 g C m-2 yr-1), which was mainly caused by warming. This study provides a new method for quantifying the contribution of ecological restoration to SOC changes, and also cautions the potential risk of LP SOC loss in the mature forest soil under future warming.

Increasing Taxa Sampling Provides New Insights on the Phylogenetic Relationship Between Eriobotrya and Rhaphiolepis.

Eriobotrya (Rosaceae) is an economically important genus with around 30 species. It is widely distributed in tropical and warm temperate regions of Asia, with most of its species in China, Myanmar, and Vietnam. However, Eriobotrya is often confused with the smaller genus Rhaphiolepis, and the phylogenetic relationships between the two genera are controversial. Here we present phylogenetic analyses of 38 newly generated Eriobotrya and Rhaphiolepis nrDNA together with 16 sequences of nrDNA and 28 sequences of ITS obtained from GenBank, representing 28 species of Eriobotrya and 12 species of Rhaphiolepis, in order to reconstruct highly supported relationships for the two genera. Contrary to previous research based on limited sampling, our results highlight the monophyly of Eriobotrya as well as Rhaphiolepis. The topology recovered here is consistent with key morphological synapomorphies such as the persistent sepals in Eriobotrya. Our findings show that increased sampling of taxa can provide a more robust phylogeny through reducing phylogenetic error and increasing overall phylogenetic accuracy.