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All biological functions evolve by fixing beneficial mutations and removing deleterious ones. Therefore, continuously fixing and removing the same essential function to separately diverge monophyletic gene families sounds improbable. Yet, here we report that brassinosteroid insensitive1 kinase inhibitor1 (BKI1)/membrane-associated kinase regulators (MAKRs) regulating a diverse function evolved into BKI1 and MAKR families from a common ancestor by respectively enhancing and losing ability to bind brassinosteroid receptor brassinosteroid insensitive1 (BRI1). The BKI1 family includes BKI1, MAKR1/BKI1-like (BKL) 1, and BKL2, while the MAKR family contains MAKR2-6. Seedless plants contain only BKL2. In seed plants, MAKR1/BKL1 and MAKR3, duplicates of BKL2, gained and lost the ability to bind BRI1, respectively. In angiosperms, BKL2 lost the ability to bind BRI1 to generate MAKR2, while BKI1 and MAKR6 were duplicates of MAKR1/BKL1 and MAKR3, respectively. In dicots, MAKR4 and MAKR5 were duplicates of MAKR3 and MAKR2, respectively. Importantly, BKI1 localized in the plasma membrane, but BKL2 localized to the nuclei while MAKR1/BKL1 localized throughout the whole cell. Importantly, BKI1 strongly and MAKR1/BKL1 weakly inhibited plant growth, but BKL2 and the MAKR family did not inhibit plant growth. Functional study of the chimeras of their N- and C-termini showed that only the BKI1 family was partially reconstructable, supporting stepwise evolution by a seesaw mechanism between their C- and N-termini to alternately gain an ability to bind and inhibit BRI1, respectively. Nevertheless, the C-terminal BRI1-interacting motif best defines the divergence of BKI1/MAKRs. Therefore, BKI1 and MAKR families evolved by gradually gaining and losing the same function, respectively, extremizing divergent evolution and adding insights into gene (BKI1/MAKR) duplication and divergence.

Puerarin can protect chondrocytes, whereby ameliorating osteoarthritis. Puerarin also promotes autophagy. Autophagy maintains chondrocyte homeostasis. The role of autophagy in puerarin-protected chondrocytes is unknown. Puerarin promoted chondrocyte autophagy. Puerarin-protected chondrocytes were reversed by autophagy inhibitors and Beclin1 inhibitor. 3-MA or Beclin1 inhibitor in vivo reversed puerarin-ameliorated cartilage damage of osteoarthritis mice. Thus, puerarin can protect chondrocytes through Beclin1-dependent autophagy activation.

Brassinosteroids (BRs) are essential plant hormones. In angiosperms, brassinolide and castasterone, the first and second most active BRs, respectively, are synthesised by CYP85A2 and CYP85A/A1, respectively. BRs in angiosperms function through an essential receptor, BR Insensitive 1 (BRI1). In addition, some angiosperms also have non-essential BRI1-like 1/3 (BRL1/3). In conifers, BRs promote seed germination under drought stress; however, how BRs function in gymnosperms is unknown. In this study, we performed functional complementation of BR biosynthesis and receptor genes from Picea abies with respective Arabidopsis mutants. We found that P. abies possessed functional PaCYP85A and PaBRL1 but not PaCYP85A2 or PaBRI1, and this results in weak BR signaling, and both PaCYP85A and PaBRL1 were abundantly expressed. However, neither BR treatment of P. abies seedlings nor expression of PaBRL1 in the Arabidopsis Atbri1 mutant promoted plant height, despite the fact that BR-responsive genes were activated. Importantly, chimeric AtBRI1 replaced with the BR-binding domain of PaBRL1 complemented the Atbri1 phenotypes. Furthermore, PaBRL1 had less kinase activity than BRI1 in vitro. Overall, P. abies had weak but still active BR signaling, explaining aspects of its slow growth and high stress tolerance. Our study sheds light on the functional and evolutionary significance of distinct BR signaling that is independent of BRI1 and brassinolide.

To compare the cervical sagittal balance and surgical outcomes between anterior cervical discectomy and fusion (ACDF) and hybrid decompression and fusion (HDF; 1-level corpectomy combined with 1-level discectomy) for consecutive 3-level cervical spondylotic myelopathy (CSM).From January 2013 to June 2016, 82 patients with 3-level CSM who underwent ACDF (n = 40) and HDF (n = 42) were retrospectively reviewed. Perioperative parameters, clinical outcomes, and radiologic sagittal alignment were analyzed and compared.Patients were followed up for 35.5 ± 6.5 months (range, 25-53 months). All patients had achieved significant improvement in Neck Disability Index and Japanese Orthopedic Association scores after operation, with similar clinical outcomes between both groups (P0.05). In the ACDF group, 2 patients were found with axial symptoms, and 1 with hoarseness. In the HDF group, 5 patients were found with axial symptoms, 1 with hoarseness, 1 with dysphagia, and 1 with pseudarthrosis. The ACDF group had less operation time and bleeding compared with the HDF group (P0.05). The restoration of segmental and C2-7 lordosis were significantly greater in the ACDF group than the HDF group (P0.05). The C2-7 sagittal vertical axis and T1 slope minus C2-7 lordosis decreased in the ACDF group at final follow-up (P0.05); however, there was no obvious change in those of the HDF group (P0.05).Both ACDF and HDF were safe and effective for the treatment of 3-level CSM. ACDF showed superiority to HDF in terms of less blood loss, shorter operation time, and better postoperative sagittal balance.

Brassinosteroid (BR) signaling has been identified from the ligand BRs sensed by the receptor Brassinosteroid Insensitive 1 (BRI1) to the final activation of Brassinozole Resistant 1/bri1 EMS-Suppressor 1 through a series of transduction events. Extensive studies have been conducted to characterize the role of BR signaling in various biological processes. Our previous study has shown that Excess Microsporocytes 1 (EMS1) and BRI1 control different aspects of plant growth and development via conserved intracellular signaling. Here, we reveal that another receptor, NILR1, can complement the bri1 mutant in the absence of BRs, indicating a pathway that resembles BR signaling activated by NILR1. Genetic analysis confirms the intracellular domains of NILR1, BRI1 and EMS1 have a common signal output. Furthermore, we demonstrate that NILR1 and BRI1 share the coreceptor BRI1 Associated Kinase 1 and substrate BSKs. Notably, the NILR1-mediated downstream pathway is conserved across land plants. In summary, we provide evidence for the signaling cascade of NILR1, suggesting pan-brassinosteroid signaling initiated by a group of distant receptor-ligand pairs in land plants.

Biofuel crops are one of the most promising regenerative alternatives of energy resources to fossil fuels. Revealing the current and future resource distribution patterns of biofuel crops will promote the development of green energies and the mitigation of greenhouse gas emissions. In this study, we first conducted a comprehensive and systematic analysis on the distribution patterns of main biofuel crops in China, using 22,352 occurrence records of 31 biofuel plant species and thirty-year environmental variables (1970-2000) with maximum entropy modeling, as well as nine-year field investigation of land use (2011-2019). The results showed that there were six different sub-regions for main biofuel crops in China, while Southwest China and South China were determined as the main concentrated potential regions. Specifically, the ranges of these regions were wider than those of current land use of main biofuel crops in China, indicating great potential for industrial cultivation. Moreover, the main biofuel crops had diverse changing patterns including increase, decrease and unstable under future climate change. Among them, biofuel crops with increase pattern (six crops) and decrease pattern (seven crops) should receive high attention for future resource utilization and production. Further field validation results confirmed that the above distribution patterns were mainly determined by increasing global temperature and precipitation. Collectively, these results will provide valuable references for the utilization and development of main biofuel resources under climate change in China, thereby shedding light on studies regarding the production of green biofuels globally.

Background: In recent years, patent foramen ovale (PFO) has been reported to be strongly associated with embolic strokes of undetermined source (ESUS), including cryptogenic stroke, transient ischemic attack (TIA), migraine, and so on.The aim of this research was to compare the sensitivity and positive predictive value of contrast transcranial Doppler (c-TCD), contrast- transthoracic echocardiography (c-TTE) versus contrast- transesophageal echocardiography (c-TEE), to find which is the best method to diagnose patent foramen ovale (PFO), provides reference for the further improvement of clinical. Methods: We investigated 161 patients who suffered from migraine, cryptogenic stroke, TIA, and cerebral infarction of unknown cause, all patients underwent the transcatheter examination, and put the results of the right heart catheterization (RHC) as the gold standard for PFO diagnosis. A chi-square test was used to compare the sensitivity and specificity between the three methods. A P value of Results: The present study revealed that c-TTE with the Valsalva maneuver yielded a higher sensitivity in detecting PFO-RLS, but the rate of misdetection might be higher than c-TCD. Conclusions: For the suspected patients, can examine with c-TCD first, if we get the positive results, then c-TTE and c-TEE should be taking to further confirmed. These findings may be helpful in the diagnosis of patent foramen ovale in practice.

Developing novel processing approaches is an effective way to improve the industrial utilization and production of crops. Dogwood (Cornus officinalis Sizb. et Zucc.) is an industrial crop in the Cornaceae family, which has been used as a traditional Chinese medicine with good medicinal values. Nevertheless, little is reported on the processing technology of C. officinalis products. In this study, we conducted a comprehensive study on the development and optimization of novel processing methods of C. officinalis fruits. The processing approach of fermentation was first used to obtain fruit extracts of C. officinalis. The fresh fruits and dried fruits were used as raw materials of fermentation. Three different parameters including sugar content, SO2 addition and yeast inoculation were chosen as variables to determine the best fermentation condition using response surface methodology (RSM). The contents of total organic acids, total iridoid glycosides, and seven medicinal active ingredients were estimated to reveal the accumulation change of bioactive products during processing. The results showed that the main-fermentation time of fresh fruits is 12–16 days, and the after-fermentation time is 30–35 days, while the main-fermentation time of dried fruits is 16–20 days, and the after-fermentation time is 30–35 days. Compared with dry-fruit extracts, fresh-fruit extracts have better quality with higher contents of medicinal active ingredients. Collectively, these results provide valuable references for the processing of C. officinalis products. Moreover, the novel processing approaches of fermentation proposed in this study will shed light on further development of other medicinal crops.

Drought is one the critical abiotic factors that reduces the germination, growth and yield of crops. Therefore the present project was designed with the objective to screen the best drought tolerant hybrid of Vigna radiata. Genetic variations for drought tolerance among these hybrids were assessed by simple and efficient technique. Seven hybrids of V. radiata (9801, 7002, 9706, 08003, 07007, 97012 and 08007) were used for screening against three levels of drought stress (zero, control), 5% (-0.05 MPa) and 10% (-0.1 MPa) induced by Polyethyleneglycol (PEG6000). A higher proline content was observed in 07007 (6.10 μg/g fresh weight) as compared to all other hybrids. Treated seedlings of each hybrid were compared with their respective control to evaluate the differences in their growth under drought. Different parameters such as percentage of germination, germination stress tolerance index (GSI), shoot and root weights (bath fresh and dry) and lengths, root length stress index (RLSI), dry matter stress index (DMSI) and plant height stress index (PHSI) showed considerable variations. Germination percentage, shoot weights, PHSI and DMSI decreased in all hybrids along with the increase of PEG induced drought stress (5% and 10% PEG). In contrary, root weights and RLSI were increased under drought. Overall 07007 showed a better performance, and can therefore be classified as a drought tolerant hybrid.

Environmental gradients play important roles in shaping the diversity of functional traits of crops. Therefore, it is imperative to reveal how crops respond to environmental shifts. In this study, a ten-year investigation (from 2008 to 2018) of an important medicinal crop, dogwood (Cornus officinalis Sieb. et Zucc.), was conducted to reveal the effects of environmental shifts on crop functional traits and bioactive products. The C. officinalis plants originated from three main producing areas were cross-regionally introduced to study area. The key environmental factors of the three original regions (ORs) and the introduced region (IR) were monitored. The functional traits and bioactive products of C. officinalis were estimated. Furthermore, the correlations among functional traits, bioactive products, and environmental factors were analyzed. The results showed that there were significant differences between functional traits and bioactive products of C. officinalis in ORs and in IR (P

Revealing the potential geographical distribution and regions that yield maximum harvest of products will be of great significance for evaluation and utilization of crops in medicinal industry. Diabetes has become a global health problem that threatens almost all nations in the world. The traditional Chinese medicine (TCM) has long been used for the treatment of diabetes in history. Specifically, four medicinal plants including huangqi (Astragalus membranaceus [Fisch.] Bunge.), huanglian (Coptis chinensis Franch.), shengdi (Rehmannia glutinosa [Gaetn.] Libosch. ex Fisch. et Mey.), and xuanshen (Scrophularia ningpoensis Hemsl.) have been confirmed effective in diabetes treatment with clear anti-diabetic mechanisms. In recent years, increasing demand of pharmaceutical industry as well as human activities lead to the degradation of habitat and drop in production of the four species. A pressing problem faced by scientists is how to propose effective conservation and utilization strategies of these medicinal crops. In this study, a comprehensive evaluation on the potential geographical distribution of the four medicinal crops was conducted based on the maximum entropy (Maxent) modeling. A total of 227, 426, 433, and 353 localities for A. membranaceus, C. chinensis, R. glutinosa, and S. ningpoensis, respectively, in China were used as the occurrence data. Ten bioclimatic variables were used as environmental factors. The results showed that the Maxent modeling generated high area under curve (AUC) values of 0.947, 0.966, 0.959, and 0.959, respectively. The areas of suitable distribution for the four species are 878,181 km2, 684,406 km2, 768,132 km2, 714,267 km2, respectively. The main decisive environmental factors that determine the potential distribution are bio12, bio12, bio6, and bio6, respectively. By further determining the core distribution regions and main producing provinces of the four species, it was found that A. membranaceus and R. glutinosa shared similar potential habitats which mainly located in northern China, while C. chinensis and S. ningpoensis shared similar potential habitats which mainly located in South and Central China. Furthermore, the former two species have similar suitable precipitation condition, while the latter two species have similar suitable precipitation condition. These results thus raised the possibility of conducting concentrated conservation and utilization of the four species in overlapping distribution regions to obtain the maximum products for pharmaceutical industry. Collectively, these results will provide valuable references for the conservation and utilization of the four medicinal crops as well as other endangered species.

Degradation and loss of species' suitable habitats in response to global warming are well documented, which are assumed to be affected by increasing temperature. Conversely, habitat increase of species is little reported and is often considered anomalous and unrelated to climate change. In this study, we first revealed the climate-change-driven habitat shifts of six endangered wetland plants - Bruguiera gymnorrhiza, Carex doniana, Glyptostrobus pensilis, Leersia hexandra, Metasequoia glyptostroboides, and Pedicularis longiflora. The current and future potential habitats of the six species in China were predicted using a maximum entropy model based on thirty-year occurrence records and climate monitoring (from 1960 to 1990). Furthermore, we observed the change of real habitats of the six species based on eight-year field observations (from 2011 to 2019). We found that the six species exhibited three different patterns of habitat shifts including decrease, unstable, and increase. The analysis on the main decisive environmental factors showed that these patterns of habitat shifts are counter to what would be expected global warming but are mostly determined by precipitation-related environmental factors rather than temperature. Collectively, our findings highlight the importance of combining multiple environmental factors including temperature and precipitation for understanding plant responses to climate change.

Revealing the dynamic accumulation and synthesizing mechanism of medicinal active ingredients will be of great significance for the evaluation and utilization of crops in medicinal industry. Dogwood (Cornus officinalis Sizb. et Zucc.), a perennial deciduous tree in the Cornaceae family, possesses important medicinal values in protecting nerves, treating diabetes and kidney diseases, and preventing inflammation and apoptosis. Dengtaishu (Cornus controversa Hemsl.) is another perennial tree in the Cornaceae family. As a Cornus plant that is phylogenetically close to C. officinalis, it is widely used as horticultural landscape plants. Revealing the difference of active ingredients between the two plants within the same genus as well as its molecular mechanism will help improve resource utilization of medicinal crops. In this study, a systematic analysis on the fruit characters, content of active ingredients and expression of key enzyme genes of C. officinalis and C. controversa was conducted. First, the main fruit characters and active ingredients in fruits and leaves of C. officinalis and C. controversa during different growth stages were determined and analyzed. The key enzyme genes geraniol 10-dehydrogenase (G10H) and secologanin synthase (SLS) in the loganin synthesis pathway of C. officinalis and C. controversa were cloned, and the expression level of G10H and SLS was determined by quantitative real-time PCR (qRT-PCR) in leaves and fruits of the two species at different growth stages. Finally, correlation analysis between the content of loganin and the expression of G10H and SLS was conducted to reveal the possible molecular mechanism of loganin synthesis. The results showed that: (1) The fruit length, seed length, and 100-fruit weight of C. officinalis were significantly higher than those of C. controversa (P

Scutellaria viscidula Bunge (Labiatae), a perennial herb, is an important medicinal plant that possesses broad pharmacological actions. S. viscidula contains flavonoids with good bioactivities (e.g., baicalin, wogonoside, baicalein, and wogonin) mainly in its dry root, which is used as alternative to Scutellaria baicalensis in the north of China. Furthermore, S. viscidula also has flavones with interesting diverged structures such as panicolin, viscidulin I, viscidulin II, and viscidulin III. Tracing the dynamic process of gene expression will help reveal the mechanism of flavonoid synthesis in S. viscidula, as well as the 4′-deoxyflavone biosynthesis in S. baicalensis. One way is to generate and analyze the expressed sequence tags (ESTs). However, little is known on the transcriptome information of S. viscidula, particularly the key genes involved in flavonoid biosynthesis. In this study, we conducted de novo transcriptome analysis of S. viscidula and obtained 42,310,834 reads and 40,052 unigenes, respectively. We revealed 177 genes relating to flavonoid biosynthesis, where 23 key enzyme-encoding genes including CHS, CHI, F3H, PAL, and 4CL were annotated. Furthermore, we investigated the dynamic expression of SvCHS, SvCHI, SvF3H, SvMYB2, and SvbHLH of stem, root, and leaf of S. viscidula in May, July, and September. Our results showed that these key genes had important regulatory function and exhibited positive correlation with total flavonoid content in different growth stages of S. viscidula. Collectively, this study provides high-quality transcriptome data of S. viscidula, and further gives significant information for understanding the molecular mechanism of gene expression and active ingredients in Scutellaria plants.

Cornus officinalis Sieb. et Zucc. is a small tree that is recognized with notable medicinal, economic, and ecological values. It has been used as traditional Chinese medicine (TCM) for thousands of years in China. Modern pharmacological research has revealed that cornel iridoid glycosides (CIGs, e.g., loganin and morroniside) in dried pericarp of C. officinalis have significant medicinal activities for strengthening immune functions. However, little is known on the molecular processes responsible for the production of these compounds. This is partly due to the absence of genomic resources, such as sequences of key enzyme genes in the biosynthetic pathways. In the present study, the transcriptome of C. officinalis was analyzed by the RNA sequencing. A total of 54,827 unigenes were yielded by de novo assembly, of which 31,780 unigenes were successfully annotated. As potential molecular markers, 121, 118, 96, 89, and 82 transcription factors belonged to bHLH, MYB, PHD, WRKY, and AP2-ERF were obtained, respectively. Moreover, the results showing that geraniol 10-hydroxylase (G10H) and secologanin synthase (SLS) were differentially expressed in fruits and leaves during different growing stages were confirmed by qRT-PCR. Furthermore, we identified two distinct expression patterns of G10H and SLS in loganin synthesis of C. officinalis fruits. Collectively, the genomic information and gene expression results presented in this study will be helpful for future studies on gene discovery and molecular process of loganin synthesis in C. officinalis.

Cornus officinalis is a small deciduous tree with significant medicinal values. To explore the ecological effects on phenotypic, cytological and biochemical diversity of medicinal plants, we estimated the loganin content, C-values and fruit and seed phenotypic traits of 80 C. officinalis germplasms collected from across three genuine producing areas of China and Madison, USA. Our results showed that most fruit and seed phenotypic traits of C. officinalis germplasms from Madison, USA were significantly larger than those from China, but the loganin contents and 2C values of germplasms from China (0.9% and 5.92 pg) were significantly higher than those from Madison (0.6% and 5.29 pg) ( P C. officinalis . Based on the above results, we found that C. officinalis had a tendency to decline in genome size (GS) and loganin content during the long period of adaptation and evolution with new ecological variables. We are able to presume that the ecological variable is the key factor in determining phenotypic diversity, GS and active ingredient of medicinal plants.

Destruction in wild resource and decline in genetic diversity of Scutellaria baicalensis are becoming a big problem urgently needed be dealt in China. To help efficiently utilize and conserve this medicinal plant resources, ISSR analysis was employed to reveal genetic diversity of 107 S. baicalensis accessions consisting of 82 wild and 25 cultivated germplasm resources from 6 populations in two main producing provinces in China. Only eighteen ISSR primers that resulted in accuracy and credibility, high polymorphism, well stability PCR products were selected. The maximal and minimal bands resulted from an ISSR primer are 35 and 18 respectively with a total of 480 bands were resolved in the agarose gels with an average of two alleles per locus. As indicated by the index of genetic diversity (Na = 1.96, Ne = 1.39, He = 0.25, I = 0.39), there were richness of genetic diversity in the collected samples and that the S. baicalensis germplasm had a large representativeness as primary collection. Cluster analysis by UPGMA and STRUCTURE placed the 107-germplasm resources into three linkage clusters. The samples from all the collection locations were clustered together, but the populations from one province were not clustered together. Compared to the RS and SW strategies, the LDSS proved to be more representative for core collection construct of S. baicalensis and the best sampling ratio was 10.3 % of the total germplasms. Finally, we found that the mini core collection by LDSS was composed of 11 accessions with high genetic diversity (Na = 1.82, Ne = 1.40, He = 0.25, I = 0.39). To verify the reliability of sampling strategy, the t test was used to evaluate the representativeness between core collections (constructed by LDSS), primary and reserve collection. Our results showed that no significant difference was found in the two important genetic parameters He and I (P > 0.05), thus suggesting that the LDSS will provide a rational sampling strategy for constructing a core collection of S. baicalensis germplasm resources in the future.

Scientific prediction of suitable cultivation regions is an effective way for the assessment of habitat suitability and resource conservation to protect endangered medicinal plants. In recent years, the natural habitat of Scutellaria baicalensis Georgi has been degenerating and disappearing in China owing to excessive market demand of medicinal plant resource. This paper reports a new approach to predict potential suitable cultivation regions and to explore the key environmental factors affecting the content of active ingredients in S. baicalensis using integrated Maxent (maximum entropy) modeling and fuzzy logics. The modeling procedure used 275 occurrence records and baicalin contents of S. baicalensis collected through 2000–2014, and 16 Worldclim environmental factors as well as HWSD soil data. The result showed that six environmental factors (alt, prec7, prec1, bio4, bio1 and t_ph) were determined as key influential factors that mostly affect both the habitat distribution and baicalin content of S. baicalensis. The highly suitable cultivation regions of S. baicalensis mainly distribute (with probability ≥0.50) in the northeast, the north-central and the northwest of China (total 419,857 km2). The statistically significant AUC (area under the curve) value (0.952) of ROC (receiver operating characteristic) curve indicated that integrated Maxent modeling and fuzzy logics could be used to predict the potential suitable cultivation regions of medicinal plants. These results could pave the road for the habitat conservation and resource utilization of endangered medicinal plants.

Cornus officinalis is plant species that is widely used in traditional Chinese medicine. To help the efficient utilization and conservation of this species, the genetic diversity of 73 germplasm resources collected from Zhejiang, Henan and Shaanxi provinces in China were used in a primary core collection. The germplasm resources were classified nine types and the analysis of morphological traits and chemical components showed wide variations among the germplasm types. The primary core collection of C. officinalis was evaluated by using 18 inter-simple sequence repeat (ISSR) markers and a mini core collection was developed based on three strategies on construction of a core germplasm resources. The ISSR results showed that a total of 233 alleles were identified in these 73 accessions, the genetic diversity (P = 91.02%, H = 0.293, I = 0.446) revealed with an average of 2 alleles per locus. The genetic diversity index analysis suggested that there is a great richness and uniqueness of genetic variation in this primary core collection. Cluster analysis by UPGMA and STRUCTURE placed the 73-germplasm resources into four linkage clusters. Almost all samples from one sampling location were clustered together. Compared to the RS and SW strategy, the LDSS strategy proved to be more representative in the core collection of C. officinalis. Finally, a mini core collections by LDSS, composed of 18 accessions (24.66% of the primary core collection), let more than 94.85% of polymorphic loci remained, 97.55% of the observed number of alleles and 99.28% of the effective number of alleles of the mini core collection, respectively. The Nei’s gene diversity and Shannon’s Information index of core collection were 0.294 and 0.443, respectively. Taken all together, the results indicated that mini core collections constructed by the LDSS strategy represented their primary core collections and provided a rational framework for intensive surveys of natural variation in C. officinalis genetic resources.

To study the effectiveness of long segment fixation combined with vertebroplasty (LSF-VP) for severe osteoporotic thoracolumbar compressive fractures with kyphosis deformity.Between March 2006 and May 2012, a retrospective analysis was made on the clinical data of 48 cases of severe osteoporotic thoracolumbar compressive fractures with more than 50% collapse of the anterior vertebral body or more than 400 of sagittal angulation, which were treated by LSF-VP in 27 cases (LSF-VP group) or percutaneous kyphoplasty (PKP) in 21 cases (PKP group). All patients suffered from single thoracolumbar vertebral compressive fracture at T11 to L2. There was no significant difference in gender, age, spinal segment, and T values of bone mineral density between 2 groups (P0.05). The effectiveness of the treatment was appraised by visual analogue scale (VAS), Cobb angle of thoracolumbar kyphosis, height of anterior/posterior vertebral body, and compressive ratio of vertebrae before and after operations.The LSF-VP group had longer operation time, hospitalization days, and more bone cement injection volume than the PKP group, showing significant differences (P0.05). Intraoperative blood loss in LSF-VP group ranged from 220 to 1,050 mL (mean, 517 mL). No pulmonaryor cerebral embolism or cerebrospinal fluid leakage was found in both groups. Asymptomatic bone cement leakage was found in 3 cases of LSF-VP group and 2 cases of PKP group. The patients were followed up for 16-78 months (mean, 41.1 months) in LSF-VP group, and 12-71 months (mean, 42.1 months) in PKP group. No fixation failure such as loosened or broken pedicle screw was found in LSF-VP group during the follow-up, and no re-fracture or adjacent vertebral body fracture was found. Two cases in PKP group at 39 and 56 months after operation respectively were found to have poor maintenance of vertebral height and loss of rectification (Cobb angle was more than 40 degrees) with recurrence of pain, which were treated by second surgery of LSF-VP; another case had compressive fracture of the adjacent segment and thoracolumbar kyphosis at 16 months after operation, which was treated by second surgery of LSF-VP. There were significant differences in the other indexes between each pair of the three time points (P0.05), except the Cobb angle of thoracolumbar kyphosis, and the height of posterior vertebral body between discharge and last follow-up in LSF-VP group, and except the Cobb angle of thoracolumbar kyphosis and compressive ratio of bertebrae between discharge and last follow-up in PKP group (P0.05). After operation, the other indexes of LSF-VP group were significantly better than those of PKP group at each time point (P0.05), except the VAS score and the height of posterior vertebral body at discharge (P0.05).The effectiveness of LSF-VP is satisfactory in treating severe osteoporotic thoracolumbar compressive fractures with kyphosis deformity. LSF-VP can acquire better rectification of kyphosis and recovery of vertebral body height than PKP.

To study the osteogenic effects of a new type of peptides anchored aminated-poly-D, L-lactide acid (PA/PDLLA) scaffold in repairing femoral defect in rats.The PDLLA scaffolds were treated by ammonia plasma and subsequent anchor of Gly-Arg-Gly-Asp-Ser (GRGDS) peptides via amide linkage formation. Thus PA/PDLLA scaffolds were prepared. The bone marrow was harvested from the femur and tibia of 4 4-week-old Sprague Dawley (SD) rats, and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured by whole bone marrow adherence method. BMSCs-scaffold composites were prepared by seeding osteogenic-induced BMSCs at passages 3-6 on the PA/PDLLA and PDLLA scaffolds. The right femoral defects of 8 mm in length were prepared in 45 adult male SD rats (weighing, 350-500 g) and the rats were divided into 3 groups (n=15) randomly. BMSCs-PA/PDLLA (PA/PDLLA group) or BMSCs-PDLLA (PDLLA group) composites were used to repair defects respectively, while defects were not treated as blank control (blank control group). General state of the rats after operation was observed. At 4, 8, and 12 weeks after operation, general, radiological, histological, micro-CT observations and real-time fluorescent quantitative PCR were performed.Two rats died after operation, which was added; the other rats survived to the end of the experiment. At each time point after operation, general and radiological observations showed more quick and obvious restoration in PA/PDLLA group than in PDLLA group; no bone repair was observed in blank control group. The X-ray scores were the highest in PA/PDLLA group, higher in PDLLA group, and the lowest in blank control group; showing significant difference in multiple comparison at the other time (P0.05) except between blank control group and PDLLA group at 4 weeks (P0.05). The X-ray scores showed an increasing trend in PDLLA group and PA/PDLLA group with time (P0.05). Histological and micro-CT observations showed the best osteogenesis in PA/PDLLA group, better in PDLLA group, and worst in blank control group. Comparison between groups had significant differences (P0.05) in bone mineral density, bone volume/total volume of range of interest, trabecular number, and structure model index. Significant differences (P0.05) were found in the expression levels of osteogenesis-related genes, such as osteocalcin, alkaline phosphatase, collagen type I, bone morphogenetic protein 2, and osteopontin when compared PA/PDLLA group with the other groups by real-time fluorescent quantitative PCR analysis.The PA/PDLLA scaffolds can accelerate the repair of femoral defects in rats.

Introduction: Conventional methods used to detect and characterize influenza viruses in biological samples face multiple challenges due to the diversity of subtypes and high dissimilarity of emerging strains. Next-generation sequencing (NGS) is a powerful technique that can facilitate the detection and characterization of influenza, however, the sequencing strategy and the procedures of data analysis possess different aspects that require careful consideration. Methodology: The RNA from the lungs of ferrets infected with influenza A/California/07/2009 was analyzed by next-generation sequencing (NGS) without using specific PCR amplification of the viral sequences. Several bioinformatic approaches were used to resolve the viral genes and detect viral quasispecies. Results: The genomic sequences of influenza virus were characterized to a high level of detail when analyzing the short-reads with either the fast aligner Bowtie2, the general purpose aligner BLASTn or de novo assembly with Abyss. Moreover, when using distant viral sequences as reference, these methods were still able to resolve the viral sequences of a biological sample. Finally, direct sequencing of RNA samples did not provide sufficient coverage of the viral genome to study viral quasispecies, and, therefore, prior amplification of the viral segments by PCR would be required to perform this type of analysis. Conclusions: the introduction of NGS for virus research allows routine full characterization of viral isolates; however, careful design of the sequencing strategy and the procedures for data analysis are still of critical importance.

Background Understanding the pathogenesis of influenza infection is a key factor leading to the prevention and control of future outbreaks. Pandemic 2009 Influenza H1N1 infection, although frequently mild, led to a severe and fatal form of disease in certain cases that make its virulence nature debatable. Much effort has been made toward explaining the determinants of disease severity; however, no absolute reason has been established. Results This study presents the heterogeneous virulence of clinically similar strains of pandemic 2009 influenza virus in human alveolar adenocarcinoma cells and mice. The viruses were obtained from patients who were admitted in a local hospital in China with a similar course of infection and recovered. The A/Nanchang/8002/2009 and A/Nanchang/8011/2009 viruses showed efficient replication and high lethality in mice while infection with A/Nanchang/8008/2009 was not lethal with impaired viral replication, minimal pathology and modest proinflammatory activity in lungs. Sequence analysis displayed prominent differences between polymerase subunits (PB2 and PA) of viral genomes that might correlate with their different phenotypic behavior. Conclusions The study confirms that biological heterogeneity, linked with the extent of viral replication, exists among pandemic H1N1 strains that may serve as a benchmark for future investigations on influenza pathogenesis.

Introduction: South China has a proven role in the global epidemiology of previous influenza outbreaks due to its dual seasonal pattern. We present the virologic, genetic and clinical characterization of pandemic H1N1 influenza infection (pH1N1) in Shantou and Nanchang, cities in southern China, during the second wave of the 2009-2010 pandemic. Methodology: Nasopharyngeal swabs were collected from 165 individuals with influenza-like illness (ILI) who presented to the hospitals in Shantou and Nanchang. Laboratory diagnosis and characterization was performed by real-time PCR, virus isolation in embryonated chicken eggs, and sequencing. Results: pH1N1 activity was sustained in three different temporal patterns throughout the study period. The overall positivity rate of pH1N1 was 50% with major distribution among young adults between the ages of 13 and 30 years. High fever, cough, expectoration, chest pain, myalgia, nasal discharge and efficient viral replication were observed as major clinical markers whereas a substantial number of afebrile cases (17%) was also observed. Rate of hospitalization and disease severity (39%) and recovery (100%) were also high within the region. Furthermore, severe complications were likely to develop in young adults upon pH1N1 infection. Genetic characterization of the HA and NA genes of pH1N1 strains exhibited homogenous spread of pH1N1 strains with 99% identity with prototypic strains; however, minor unique mutations were also observed in the HA gene. Conclusion: The study illustrates the detailed characteristics of 2009 influenza pandemic in southern parts of China that might help to strategize preparedness for future pandemics and subsequent influenza seasons.