Low-level Cu-fortification of bovine lactoferrin: Focus on its effect on in vitro anti-inflammatory activity in LPS-stimulated macrophages.

Bovine lactoferrin (LF) per 1 g was reacted with 0.16, 0.32, and 0.64 mg CuCl2 to reach 10%, 20%, and 40% copper-saturation, respectively, aiming to assess their anti-inflammatory activities to lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. The macrophages treated with CuCl2 at 0.051 µg/mL dose did not have obvious change in cell viability, lactate dehydrogenase (LDH) release, and intracellular reactive oxygen species (ROS) production. However, LF and Cu-fortified LF products (10-80 µg/mL doses) mostly showed inhibitory effects on the stimulated macrophages dose-dependently. Moreover, Cu-fortified LF products of lower Cu-fortifying levels at lower doses exerted weaker inhibition on the stimulated macrophages than LF, leading to higher cell viability but decreased LDH release. Meanwhile, LF and Cu-fortified LF products at 10 and 20 µg/mL doses showed different activities to the stimulated cells, via partly decreasing or increasing the production of inflammatory mediators namely prostaglandin E2 (PGE2), nitric oxide, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1ß, and ROS production, depending on the used Cu-fortifying and dose levels. Compared with LF, Cu-fortified LF product (Cu-fortifying level of 0.16 mg/g LF) at 10 µg/mL dose showed enhanced inhibition on the production of PGE2, ROS, IL-1ß, and TNF-α, evidencing increased anti-inflammatory activity. However, the inhibition of Cu-fortified LF product (Cu-fortifying level of 0.32 mg/g LF) at 20 µg/mL dose on the production of these inflammatory mediators was mostly reduced. It is thus proposed that both Cu-fortifying and dose levels could affect LF's anti-inflammatory activity in LPS-stimulated macrophages, while the Cu-fortifying level of LF could govern activity change.

Integration of Artificial Intelligence, Blockchain, and Wearable Technology for Chronic Disease Management: A New Paradigm in Smart Healthcare.

Chronic diseases are a growing concern worldwide, with nearly 25% of adults suffering from one or more chronic health conditions, thus placing a heavy burden on individuals, families, and healthcare systems. With the advent of the "Smart Healthcare" era, a series of cutting-edge technologies has brought new experiences to the management of chronic diseases. Among them, smart wearable technology not only helps people pursue a healthier lifestyle but also provides a continuous flow of healthcare data for disease diagnosis and treatment by actively recording physiological parameters and tracking the metabolic state. However, how to organize and analyze the data to achieve the ultimate goal of improving chronic disease management, in terms of quality of life, patient outcomes, and privacy protection, is an urgent issue that needs to be addressed. Artificial intelligence (AI) can provide intelligent suggestions by analyzing a patient's physiological data from wearable devices for the diagnosis and treatment of diseases. In addition, blockchain can improve healthcare services by authorizing decentralized data sharing, protecting the privacy of users, providing data empowerment, and ensuring the reliability of data management. Integrating AI, blockchain, and wearable technology could optimize the existing chronic disease management models, with a shift from a hospital-centered model to a patient-centered one. In this paper, we conceptually demonstrate a patient-centric technical framework based on AI, blockchain, and wearable technology and further explore the application of these integrated technologies in chronic disease management. Finally, the shortcomings of this new paradigm and future research directions are also discussed.

[Overexpression of Interferon Regulatory Factor IRF1 Remodels Homeostasis of K562 Cells].

OBJECTIVE: To investigate the effects of IRF1 on the homeostasis and differentiation of K562 cells. METHODS: Three different vectors were constructed to screen the best strategy for IRF1 overexpression. The effect of IRF1 on cell proliferation and apoptosis was explored by cell count and apoptotic surface marker detection. Likely, the effect of IRF1 on cell differentiation was analyzed by differentiational surface marker assay. Finally, the regulation mechanism at mRNA level was analyzed by RT-qPCR. RESULTS: The single open reading frame constructed by P2A-T2A element showed the highest expression intensity, and it was the best approach to realize IRF1 enhancement. Cell counts showed that IRF1 had no significant effect on the proliferation of K562. Annexin V and 7-AAD labeling exhibited strong anti-apoptotic function of IRF1 against AraC induction. Flow cytometry revealed that IRF1 overexpression could also further increase the proportion of CD71+CD235a+ cells. RT-qPCR confirmed its upregulation effect on CD235a and TAL1. CONCLUSION: IRF1 enhancement alters the homeostasis characteristics of K562 cells, increases the anti-apoptotic ability and raises the potential to downstream differentiation, suggesting that IRF1 may play an important regulatory role in the hematopoietic development, including erythropoiesis.

Solvothermal Synthesis of Humic Acid-Supported CeO2 Nanosheets Composite as High Performance Adsorbent for Congo Red Removal.

Surface properties and structures of materials are essential for their adsorption of pollutants in water. Humic acids (HA)-supported CeO2 nanosheet composites are synthesised by solvothermal method. The size of CeO2 nanosheets are approximately 100-500 nm. The obtained composite exhibits superior adsorption ability for Congo Red (CR) in water, which can be attributed to its unique structure and highly dispersed CeO2 nanosheet. The composite's adsorption behaviour of CR follows a pseudo-second-order mode and Langmuir adsorption model well, and the maximum adsorptive capacity for CR achieves 260 mg g-1. The presence of CeO2 nanosheets enhances surface area and enriches the mesoporous structure of the composites, thereby promoting CR adsorption capacity.

Application of contrast-enhanced ultrasonography in the diagnosis of post-kidney transplant lymphoproliferative disorder in native kidney- a case report.

BACKGROUND: Post-transplant lymphoproliferative disorders (PTLDs) represent a spectrum of heterogenetic lymphoid proliferations. PTLD is a serious complication that affects the long-term survival of kidney transplant patients. Imaging examination is an important method for detecting and diagnosing PTLD. Contrast-enhanced ultrasonography (CEUS) and CEUS-guided biopsy are important modalities for tumor detection and diagnosis. In this case, we describe a 69 years old man in whom a native kidney PTLD was confirmed by CEUS. CASE PRESENTATION: A 69-year-old male patient who had a kidney transplant 1 year earlier presented with 3 months of progressive myasthenia of both lower limbs associated with amyotrophy and weight loss. Although positron emission tomography/computed tomography (PET-CT) showed a high metabolic lesion in the untransplanted kidney, abdominal contrast enhanced computed tomography cannot detect the lesion in the atrophic left kidney. The above examinations showed that the transplanted kidney was normal. CEUS can detect a homogeneously enhanced lesion in the same location as PET-CT. Subsequently, a biopsy was performed under CEUS guidance, and the final pathological diagnosis was diffuse large B-cell lymphoma. The patient then received the R-CHOP treatment. Unfortunately, pulmonary thromboembolism occurred 2 weeks later, and the patient's condition was not alleviated through active treatment. Finally, the patient's family gave up treatment, and the patient was discharged. CONCLUSION: The case suggested that CEUS was a valuable imaging method for patient with renal transplantation to detect and diagnose of PTLD.

Comparison of the Effects of the Alcalase-Hydrolysates of Caseinate, and of Fish and Bovine Gelatins on the Acidification and Textural Features of Set-Style Skimmed Yogurt-Type Products.

Commercial caseinate and two gelatins from bovine and fish skin were hydrolyzed by alcalase, and used at 2 g/kg in skimmed bovine milk that was then fermented with a commercial direct vat set starter, to clarify different effects of these hydrolysates on acidification and textural attributes of set-style yogurt samples. Compared with the fermentation of the yogurt sample without hydrolysate addition, the two gelatin hydrolysates in the yogurt samples endowed lower titratable acidity but higher pH values and thus delayed yogurt fermentation, while the caseinate hydrolysate showed an effect opposite to the two gelatin hydrolysates. The two gelatin hydrolysates induced worse quality attributes for the resultant yogurt samples, including higher syneresis extent, smaller hysteresis loop areas, and lower values in these textural indices like hardness, adhesiveness, apparent viscosity, elastic and viscous moduli. However, the caseinate hydrolysate led to improved quality attributes. Moreover, bovine gelatin hydrolysate always had a greater negative effect than fish gelatin hydrolysate on yogurt acidification and texture. It is concluded that these gelatin hydrolysates could confer the yogurt with intended bio-activities of gelatin hydrolysates but negatively impact yogurt acidification and texture, while the caseinate hydrolysate might be helpful for yogurt processing by shortening fermentation time and improving yogurt texture.

[Comparison of Effect of Serum-Free Culture Systems on Directional Erythroid Differentiation of Human Umbilical Cord Blood CD34+ Cells].

OBJECTIVE: To compare the efficacy of directional erythroid differentiation in different serum free culture systems and to screen the optimal culture systems for inducing the differentiation of umbilical cord blood hematopoietic stem and progenior cells (HSPC) to erythroid cells. METHODS: The CD34+ cells from umbilical blood munonuclear cells were sorted by using the magnetic beads, and were inoculated into 3 different of culture systems (system 1, 2 and 3 respectively), to induce erythrold differentiation by 3 stage culture. The living cells were counted in different differentiation stages and were observed by Wright-Giemsa staining; the expression of CD71 and CD235a on cell surface was detected by flow cytometry, the erythroid differentiation pteency was detected via colony-forming test. RESULTS: The ability of system 2 to promote the HSPC proliferation was the strongest, the efficacy of system 3 to promote the erythroid differentiation of HSPC was the most optimal; the proliferation ability of cells cultured in system 2 for 2-15 days all was higher than that of cells cutured in system 1 and 3 (P＜0.05). The flow cytometry detection showed that the expression of CD71 and CD235a on surface of cells cultured in system 3 was the highest, the CD235a percentage on day 15 of differentiation in system 3 was (92.33±3.89)%, that in system 2 was (84.67±3.12)%, while that in system 1 was (72.17±6.83)% (P＜0.05). Cell morplologic detection showed that throid differentiation was accelerated on day 12, the percentage of orthochromatic erythrocytes in system 3 was (67.67±2.08)% which was 10.69 and 25.34 times higher than that in system 2 and 1 respectively (P＜0.05). The colony-forming test showed the ratio of BFU-E in system 3 increased gradually on day 3-9 (r=0.99, P＜0.05), which was significanlly higher than that in system 2 and 1 on day 9 (90.35±5.52% vs 77.06±2.26% and 74.50±3.95%). CONCLUSION: Culture system 3 is the most effective serum-free erythroid differentiation system, and the culture system 2 is the most powerful HSPC proliferation system. This study results provide a technical basis for further efficiently increasing and inducing the erythroid proliferation and differentiation of HSPC, and also provide culture system in vitro for the clinical application and basic research.

Allyl methyl trisulfide protected against acetaminophen (paracetamol)-induced hepatotoxicity by suppressing CYP2E1 and activating Nrf2 in mouse liver.

In order to investigate the protective effects of allyl methyl trisulfide (AMTS) on acetaminophen (APAP)-induced hepatotoxicity, 75 KM mice were randomized into 5 groups, i.e. a control group, an APAP group, and three AMTS/APAP groups. The mice in the AMTS/APAP groups and APAP group were gavaged with 25-100 mg kg-1 AMTS or corn oil for 7 d followed by intraperitoneal injection of 300 mg kg-1 APAP, while mice in the control group were treated with a vehicle. We found that AMTS significantly attenuated APAP-induced hepatotoxicity shown by reduced mortality, decreased serum aminotransferase activities, and improved liver histological morphology. APAP overdose resulted in a significant increase of hepatic malondialdehyde (MDA) level and a decrease of the protein levels of NQO-1, γ-GCS, HO-1, and SOD, which was suppressed by AMTS pretreatment. Furthermore, AMTS inhibited the APAP-induced elevation of hepatic p62 and LC3II protein levels. Interestingly, AMTS attenuated the APAP-induced decline of hepatic CYP2E1 protein levels, but AMTS alone led to the decrease of CYP2E1 protein expression in mouse liver. Collectively, these data suggest that AMTS could attenuate APAP-induced hepatotoxicity by suppressing CYP2E1 and activating Nrf2.

Tea Plant Information Archive: a comprehensive genomics and bioinformatics platform for tea plant.

Tea is the world's widely consumed nonalcohol beverage with essential economic and health benefits. Confronted with the increasing large-scale omics-data set particularly the genome sequence released in tea plant, the construction of a comprehensive knowledgebase is urgently needed to facilitate the utilization of these data sets towards molecular breeding. We hereby present the first integrative and specially designed web-accessible database, Tea Plant Information Archive (TPIA; http://tpia.teaplant.org). The current release of TPIA employs the comprehensively annotated tea plant genome as framework and incorporates with abundant well-organized transcriptomes, gene expressions (across species, tissues and stresses), orthologs and characteristic metabolites determining tea quality. It also hosts massive transcription factors, polymorphic simple sequence repeats, single nucleotide polymorphisms, correlations, manually curated functional genes and globally collected germplasm information. A variety of versatile analytic tools (e.g. JBrowse, blast, enrichment analysis, etc.) are established helping users to perform further comparative, evolutionary and functional analysis. We show a case application of TPIA that provides novel and interesting insights into the phytochemical content variation of section Thea of genus Camellia under a well-resolved phylogenetic framework. The constructed knowledgebase of tea plant will serve as a central gateway for global tea community to better understand the tea plant biology that largely benefits the whole tea industry.

Effect of the Zn Supplementation on Immuno-Modulatory Activities of Bovine Lactoferrin in the Murine Splenocytes and RAW264.7 Macrophages.

Lactoferrin (LF) has important bio-functions including immuno-modulation, while essential trace metals may interact with LF and thereby induce property especially bio-activity changes. Bovine LF was thus supplemented with Zn2+ at 0.16, 0.32, and 0.64 mg/g LF to yield 10%, 20%, and 40% Zn-saturation, respectively. Afterwards, bovine LF and the Zn-supplemented LF products at 10-40-µg/mL doses were compared for their immuno-modulatory activities in two immune cells (murine splenocytes and RAW264.7 macrophages), using the stimulation index of the splenocytes, T lymphocyte subpopulations, macrophage phagocytosis, and cytokine production as evaluation reflectors. The results showed that bovine LF and the Zn-supplemented LF products had suppressive effect on the splenocytes and concanavalin A (ConA)- and lipopolysaccharide-stimulated splenocytes, but lower Zn-saturation and lower dose could alleviate and even counteract this suppressive effect (P < 0.05). More importantly, the Zn-supplemented LF product with lower Zn-saturation at lower dose exerted slightly higher macrophage stimulation, increased CD4+/CD8+ ratio of T lymphocyte subpopulations, and were capable of enhancing the interleukin-2 (IL-2), IL-4, and interferon-γ production in the splenocytes or the IL-1ß, IL-6, and tumor necrosis factor-α production in the macrophages significantly (P < 0.05). Contrary to its counterpart at lower dose, the Zn-supplemented LF product with higher Zn-saturation at higher dose mostly showed opposite effects in the two cell models. It is concluded that Zn supplementation has an impact on the immuno-modulation of bovine LF, while Zn-saturation is a key factor to modulate these assessed immune activities.

Modulatory Effect of the Supplemented Copper Ion on In Vitro Activity of Bovine Lactoferrin to Murine Splenocytes and RAW264.7 Macrophages.

Bovine lactoferrin (LF) was supplemented with Cu2+ at three contents of 0.16, 0.32, and 0.64 mg/g LF, respectively. After then, LF and Cu-supplemented LF products were assessed for immuno-modulation in murine splenocytes and RAW264.7 macrophages, using dose levels of 10-40 µg/mL and four evaluation reflectors including stimulation index of splenocytes, T lymphocyte subpopulations, macrophage phagocytosis, and cytokine secretion. The results indicated that LF and Cu-supplemented LF products had suppression on splenocytes as well as concanavalin A (ConA)- or lipopolysaccharide-stimulated splenocytes; however, using lower Cu-supplementation content (i.e., 0.16 mg/g LF) and lower dose level (10 µg/mL) alleviated this suppression significantly (P < 0.05). Compared to LF, Cu-supplemented LF product of lower Cu-supplementation content at lower dose level yielded slightly enhanced macrophage stimulation, increased CD4+/CD8+ ratio of T lymphocyte subpopulations in ConA-stimulated splenocytes, and significant secretion enhancement for interleukin-2 (IL-2), IL-4, interferon-γ (in splenocytes), IL-1ß, and tumor necrosis factor-α (in macrophages) (P < 0.05). Furthermore, Cu-supplemented LF product of higher Cu-supplementation content (i.e., 0.64 mg/g LF) at higher dose level mostly showed opposite effects in the cells, in comparison with its counterpart at lower dose level. It is concluded that Cu-supplementation of LF can alleviate or increase LF's effects on the two immune cells, and moreover, Cu content of supplemented LF is a key factor that modulates these effects.

Gut microbiota alterations in moderate to severe acne vulgaris patients.

Acne vulgaris is a chronic inflammatory dermatosis affecting approximately 85% of adolescents. There are many factors contributing to the development of this ailment. A recent study indicated that gut microbiota takes part in the pathogenesis of acne. We aimed to investigate the link between acne vulgaris and gut microbiota. A total of 31 moderate to severe acne vulgaris patients and 31 healthy controls were enrolled. We collected their feces, and gut microbiota was evaluated by the hypervariable regions of 16S rRNA genes through high-throughput sequencing. We identified links between acne vulgaris and changes of gut microbiota. At the phylum level, Actinobacteria (0.89% in acne patients and 2.84% in normal controls, P = 0.004) was decreased and Proteobacteria (8.35% in acne patients and 7.01% in normal controls, P = 0.031) was increased. At the genus level, Bifidobacterium, Butyricicoccus, Coprobacillus, Lactobacillus and Allobaculum were all decreased. The observed difference in genera between acne patients and healthy controls provides a new insight into the link between gut microbiota changes and acne vulgaris risk.

Niclosamide attenuates inflammatory cytokines via the autophagy pathway leading to improved outcomes in renal ischemia/reperfusion injury.

Renal ischemia/reperfusion (I/R) injury is a debilitating condition that leads to loss renal function and damage to kidney tissue in the majority of patients with acute kidney disease. Previous studies have indicated that autophagy serves a protective function in renal I/R injury. In the present study, the effect of the anthelmintic niclosamide in the regulation of inflammatory responses in kidney I/R was investigated. A total of 40 Sprague-Dawley rats were randomly divided into the following 5 groups (n=8 in each group): Sham group; renal I/R injury; renal I/R injury plus 3­methyladenine (3­MA) treatment (15 mg/kg); renal I/R injury plus niclosamide (25 mg/kg); and renal I/R injury plus rapamycin (10 mg/kg). The expression levels of autophagy­associated proteins in kidney samples obtained from rats with I/R injury were examined using reverse transcription­quantitative polymerase chain reaction and western blotting techniques. In addition, histopathological alterations, the expression of cytokines and renal function were evaluated. Treatment with niclosamide was associated with induction of autophagy and an overall improvement in renal function. There was an increased expression of autophagosome­associated proteins, suggesting a strong correlation between autophagy and improvement of renal function. The increased levels of anti­inflammatory cytokines and decreased levels of pro­inflammatory cytokines provided additional evidence that niclosamide may be effective for the treatment of renal I/R injury. Clinical studies are required to further validate the results of the present study.

[The Research on Measurement System and Method of Tissue Optical Parameters with Wide Spectra Based on Double-Integrating-Spheres].

The measurement of tissue optical parameters is the focusing research content of Biomedical Photonics. The optical properties of human tissue are closely related to the physiological and pathological state. In recent years, the tissue imaging diagnosis and non-invasive detection of componentsbecome the hot research topics, applying the tissue optical properties especially the absorption and scattering properties. These provide the basis for the study of optical imaging and the spectrum detection of body composition etc. The Double-Integrating-Spheres (DIS) method can measure the absorption coefficient, scattering coefficient and so on in vitro tissuesimultaneously. It has the advantages of accurate, rapid, large applicable scope. The method applya standard method for measuring the optical parameters. This paper build the wide spectrum measurement system of optical parameters based on DIS and super continuum lasers. Then we analyze the transfer function, error sources and the best measuring conditions of the system. Finally we establish the correction forward model based on BP-MCML and the inverse algorithm of the optical parameters based on L-M algorithm. The optical parameters of intralipid solution in the wavelength range of 1,100~1,400 nm are measured. The experiment results show that the improved inverse algorithm is accurate. The multiple measurements standard deviation is within 3%. Compared the results of scattering coefficient and absorption coefficient at different wavelengths to the results of other research groups, the deviation is less than 3.4%.

[Study on the Determination System of Tissue Optical Properties Based on Diffuse Reflectance Spectrum].

The determination of tissue optical properties is the fundamental research field in biomedical optics. The ability to separately quantify absorption and scattering coefficients of tissue based on diffuse reflectance spectrum not only helps to gain physiological and structural properties of tissue but also provide insight into the mechanisms of tissue, which leads to the improvement in non-invasive detecting, image diagnosis and photodynamic therapy. In the paper, a flexible and rapid method is developed to extract the absorption and reduced scattering coefficients of turbid medium such as human tissue with diffuse reflectance spectrum. The diffuse reflectance spectrum is measured by the system which includes a white light source, a spectrometer, and a fiber optic probe for delivery and collection of light. The collection efficiency and system transfer function are researched based on the fiber probe geometry. This paper outlines a method based on empirical forward model and non-linear modeling inverse model to extract the optical properties from diffuse reflectance spectrum. The approach includes four steps: (1) generating diffuse reflectance spectra for training inverse model; (2) training the inverse model; (3) measuring and processing the diffuse reflectance spectra of samples; (4) predicting the optical properties of samples. Since the forward and inverse models could be regarded as non-linearity, the Artificial Neural Networks (ANN) is employed to develop the forward and inverse models. The principal component analysis (PCA) is also employed in the inverse model to decompress the data dimension and suppress the spectral noise. With a single fiber optic probe and spectroscopy system, the diffuse reflectance spectrum is measured and preprocessed. The accuracy and robustness of this method are evaluated by measuring the phantoms with a wide range of optical properties. The results indicate that the absorption and scattering coefficients could be extracted accurately by measuring the diffuse reflectance spectrum of single source-detector distance. The mean RMS percentage error is 4.58% and 7.92%, respectively. As to the application of extracting concentration of different chromosphere, it is better to include the absorption peak of every chromosphere within the measuring wavelength range. This method is valid for a wide range of optical properties with the advantage of rapid measurement and simple system setup, which is important for the clinical application.

[A prevalence investigation of blindness and vision impairment in 2009 in older adults of Dachang Blocks of Baoshan District, Shanghai, China].

OBJECTIVE: To investigate the prevalence of blindness and low vision and the leading causes of blindness in residents aged≥60 years in Dachang Blocks of Baoshan District, Shanghai, China. METHODS: A cross-sectional study was carried out by Shanghai Eye Disease Prevention & Treatment Center and the Center for Disease Control and Prevention in Baoshan District of Shanghai from October to December in 2009. Randomly cluster sampling was used to identify the adults aged≥60 years who had lived in Dachang Blocks of Baoshan District, Shanghai for more than 10 years. Presenting visual acuity (PVA) and best-corrected visual acuity (BCVA) based on autorefraction and subjective refraction were measured separately in each eye. External eye, anterior segment and ocular fundus were examined by the ophthalmologist using slit lamp-microscopes direct ophthalmoscopy and non-mydriatic digital camera. And the leading causes of visual impairment were assured. The Chi square test was used between the groups of rate comparison. RESULTS: Of 5199 enumerated subjects≥60 years of age, 87.42% (4545/5199) were examined. All subjects were urban population who were originally changed from the rural population in nearly 10 years. In this population, with best-corrected visual acuity, 30 persons were diagnosed as blindness, 145 persons were diagnosed as low vision. The prevalence of blindness and low vision were 0.67%, 3.19%, respectively. Low vision was associated with female gender. It was statistically significant difference (χ2=4.88, P<0.05). The leading causes of blindness were cataract, macular degeneration, ocular absence or atrophy, glaucoma, and diabetic retinopathy or corneal diseases. With presenting visual acuity, 39 persons were diagnosed as blindness, 401 persons were diagnosed as low vision. The prevalence of blindness and low vision were 0.86%, 8.82%, respectively. Blindness and low vision were associated with older age. The prevalence of blindness and low vision increased rapidly in aged 75 years or older people. The leading causes of blindness were cataract, uncorrected refractive error, macular degeneration, ocular absence or atrophy, glaucoma. Low vision was associated with female gender. It had statistically significant difference (χ2=13.345, P<0.01). CONCLUSIONS: In rapidly urbanized and aging community of Shanghai, cataract, uncorrected refractive error, macular degeneration were the leading causes of blindness with presenting visual acuity. The prevalence of low vision in females was higher than that of males which had statistically significant difference. These kinds of residents needed more targeted eye health education and services.

A test of seven candidate barcode regions from the plastome in Picea (Pinaceae).

DNA barcoding, as a tool for species discrimination, has been used efficiently in animals, algae and fungi, but there are still debates on which DNA region(s) can be used as the standard barcode(s) for land plants. Gymnosperms, especially conifers, are important components of forests, and there is an urgent need for them to be identified through DNA barcoding because of their high frequency of collection in the field. However, the feasibility of DNA barcoding in gymnosperms has not been examined based on a dense species sampling. Here we selected seven candidate DNA barcodes from the plastome (matK, rbcL, rpoB, rpoC1, atpF-atpH, psbA-trnH, and psbK-psbI) to evaluate their suitability in Picea (spruce). The results showed that none of them or their different combinations has sufficient resolution for spruce species, although matK+rbcL might be used as a two-locus barcode. The low efficiency of these candidate barcodes in Picea might be caused by the paternal inheritance of the chloroplast genome, long generation time, recent radiation, and frequent inter-specific hybridization aided by wind pollination. Some of these factors could also be responsible for the difficulties in barcoding other plant groups. Furthermore, the potential of the nuclear LEAFY gene as a land plant barcode was discussed.