Genome-Wide Identification of bZIP Transcription Factors in Faba Bean Based on Transcriptome Analysis and Investigation of Their Function in Drought Response.

Faba bean is an important cool-season edible legume crop that is constantly threatened by abiotic stresses such as drought. The basic leucine zipper (bZIP) gene family is one of the most abundant and diverse families of transcription factors in plants. It regulates plant growth and development and plays an important role in the response to biotic and abiotic stresses. In this study, we identified 18 members of the faba bean bZIP transcription factor family at the genome-wide level based on previous faba bean drought stress transcriptome sequencing data. A phylogenetic tree was constructed to group the 18 VfbZIP proteins into eight clades. Analysis of cis-acting elements in the promoter region suggested that these 18 VfbZIPs may be involved in regulating abiotic stress responses such as drought. Transcriptome data showed high expression of seven genes (VfbZIP1, VfbZIP2, VfbZIP5, VfbZIP7, VfbZIP15, VfbZIP17, and VfbZIP18) in the drought-tolerant cultivar under drought stress, in which VfbZIP1, VfbZIP2, and VfbZIP5 were consistently expressed as detected by quantitative real-time polymerase chain reaction (qRT-PCR) compared to the transcriptome data. Ectopic overexpression of the three VfbZIPs in tobacco, based on the potato Virus X (PVX) vector, revealed that VfbZIP5 enhanced the drought tolerance. Overexpressed VfbZIP5 in plants showed lower levels of proline (PRO), malondialdehyde (MDA), and peroxidase (POD) compared to those overexpressing an empty vector under 10 days of drought stress. Protein-protein interaction (PPI) analysis showed that VfbZIP5 interacted with seven proteins in faba bean, including VfbZIP7 and VfbZIP10. The results depict the importance of VfbZIPs in response to drought stress, and they would be useful for the improvement of drought tolerance.

Site-specific and seasonal variation in habitat use of Eurasian otters ( Lutralutra) in western China: implications for conservation.

As a top predator, the Eurasian otter ( Lutralutra) is an indicator of healthy freshwater ecosystems and a flagship species for conservation. Once widespread in China, the species is now distributed mainly in the upper reaches of the great rivers of western China. However, a lack of knowledge on local otter populations continues to hinder their conservation in China. Here, we conducted a detailed study on habitat use of Eurasian otters in Yushu City and Tangjiahe National Nature Reserve in western China using transect surveys. At both study sites, otters preferred to defecate on large rocks close to or protruding from the river and about 50 cm above the waterline. In Yushu, no spraints were found along the 5 km river bank section in the downtown area, with otters preferring sprainting sites with natural banks, riparian zones, and lower human population density. However, this pattern was not obvious at Tangjiahe, where river transformation and human disturbance are minor. Otter river use intensity was negatively correlated with elevation and human population density in Yushu in both seasons. In Tangjiahe, otter river use intensity was positively correlated with prey mass and flow rate and negatively correlated with human population in spring, but positively correlated with human population and negatively correlated with flow rate in autumn. These results reflect the flexible habitat use strategies of otters at different sites, underlining the necessity to study otters living in different regions and habitat types. We provide suggestions for river modification and call for more site-specific studies to promote otter conservation in China.

Repair of Bone Defects With Endothelial Progenitor Cells and Bone Marrow-Derived Mesenchymal Stem Cells With Tissue-Engineered Bone in Rabbits.

PURPOSE: This study aimed to investigate the repair of bone defects in rabbits with tissue-engineered bones using cocultured endothelial progenitor cells (EPCs) and bone marrow mesenchymal stem cells (BMSCs) as seeding cells. METHODS: Endothelial progenitor cells and BMSCs were isolated and purified from the peripheral blood and bone marrow, respectively, of New Zealand rabbits. The third passage of BMSCs was cultured alone or with EPCs. Cells were characterized using specific markers and then seeded on partially deproteinized biologic bones from pigs as a scaffold. The engineered bones were used to repair bone defects in rabbits. Hematoxylin and eosin and Masson staining were performed to examine vascularization and osteogenesis in the engineered bone. RESULTS: The cocultured EPCs and BMSCs grew well on the surface of the scaffold. Compared with monocultured BMSCs, cocultured EPCs and BMSCs promoted the formation of blood vessels and bone on the scaffold, in addition to accelerating the repair of bone defects. The collagen content was significantly increased in the scaffold with cocultured EPCs and BMSCs, compared with the scaffold seeded with mono-cultured BMSCs. CONCLUSIONS: Tissue-engineered bones seeded with cocultured EPCs and BMSCs may be used effectively for the repair of bone defects.

Discovery of first active breeding den of Chinese mountain cat ( Felis bieti).

In mid-September 2018, during a field survey in Chiat'ung, Sanjiangyuan (Three-River-Source) Region, Tibetan Plateau, China, we discovered the first active breeding den of the Chinese mountain cat (Felis bieti), inhabited by one adult female and two kittens. Based on fieldwork over the following months, five breeding dens were discovered, and 33 sightings were recorded. In addition, at least five individuals were confirmed to inhabit this overlooked region, and much previously unknown information concerning this cat species and its ecology was revealed for the first time.

Ultrasound diagnosis of intramural pregnancy.

Intramural pregnancy is a rare type of ectopic pregnancy, where the gestational sac is implanted inside the myometrium instead of the endometrial and fallopian tubes. Preoperative diagnosis remains very difficult. Ultrasonic findings vary according to the anatomical location or duration of pregnancy. In this study, we summarized the ultrasonic characteristics of intramural pregnancy by assessing three cases. We also propose a set of ultrasonic characteristics to facilitate differential diagnosis between intramural pregnancy and other types of ectopic pregnancy.

Three-dimensional transvaginal tomographic ultrasound imaging for cervical cancer staging.

The objective of this study was to investigate the feasibility of using 3-D transvaginal tomographic ultrasound imaging (TUI) to stage patients with cervical carcinoma. Eighty women with cervical cancer who underwent transvaginal TUI examinations were enrolled. In all patients, cancer was confirmed pre-operatively by pathologic examination. Staging on the basis of clinical features, ultrasonography and magnetic resonance imaging was performed according to the International Federation of Gynecology and Obstetrics (FIGO) staging system. Clinical, TUI and magnetic resonance imaging staging was compared with that based on histology. Depth of invasion into the stroma was measured by TUI in 52 cases and compared with pathologic results. An interclass correlation coefficient was used to analyze reproducibility. In total, all 80 patients underwent surgical treatment. The accuracy of pre-operative staging, compared with histologic findings, was 92.50% for TUI, 82.50% for magnetic resonance imaging and 78.75% for clinical examination. The mean depth of lesions as measured with TUI was 12.5 ± 6.2 mm (range: 3.5-40.0 mm), and that measured on histology was 10.5 ± 8.0 mm (range: 3.0-40.0 mm). The interclass correlation coefficient of the two methods was 0.933 (95% confidence interval: 0.887-0.961). Pre-operative TUI is promising as a method for pre-operative staging of cervical carcinomas. TUI can also reliably assess lesion depth.

[Establishment and experimental study of alveolar preservation before dental implantation in Beagle dogs].

PURPOSE: To establish the model of alveolar ridge preservation after tooth extraction for dental implant replacement, and to observe the effect of tissue engineered bone on osseointegration. METHODS: Isolated BMSCs were expanded and osteogenically induced in vitro. The tissue engineering complex was constructed with BMSCs/A-PCPC in vitro. Six extraction sockets, with three on each side, were created in the mandibles of four Beagle dogs by extracting the second, third and fourth premolars. BMSCs/A-PCPC were placed on one side of the extraction sockets, while autogenous bone, A-PCPC and nothing were placed on the other side as control. X-ray and CT scans were conducted 1day, 4 and 12 weeks after operation to detect the change of the alveolar ridge. The bone of sockets were harvested at 8-week post-implantation and subject to histological for evaluating. SPSS17.0 software package was used for data analysis. RESULTS: Radiographs demonstrated higher radiodensity in group of complex than in simple materials group, autogenous bone group after 4 weeks. Hard tissue biopsy at 12-week showed that bone activity of BMSCs/A-PCPC complex was better than the other groups. Spiral CT analysis showed that alveolar ridge of each group experienced a certain degree of absorption. At 12-week, the alveolar ridge height reduction values in A-PCPC group was smaller than in A-PCPC group, autogenous bone group and blank group (P<0.05). CONCLUSION: The BMSCs/A-PCPC complex is favorable for preservation of alveolar ridge.

Coculture of vascular endothelial cells and adipose-derived stem cells as a source for bone engineering.

The interaction between vascular endothelial cells (VECs) and osteoblasts (OBs) is the focus of this recent research. Vascular endothelial cells secrete bone morphogenetic protein, which promotes OB differentiation and stimulates OBs and their precursor cells to secrete vascular endothelial growth factor. Vascular endothelial growth factor is important in angiogenesis and angiopoiesis. Cloning studies have shown that adipose-derived stem cells (ADSCs) have the potential to differentiate into fat, bone, cartilage, and skeletal and smooth muscle cells, among others. Adipose-derived stem cells can express multiple growth factors, including vascular endothelial growth factor and hepatocyte growth factor. Our study examined the influence of coculturing VECs and ADSCs on osteogenic differentiation. Cord blood-derived VECs and ADSCs were isolated from rats and characterized with immunofluorescence staining and morphological observation. Coculture of third-generation ADSCs and VECs was induced for 6 weeks. Cell growth was analyzed using a modified MTT assay. Alkaline phosphatase (ALP) and osteocalcin (OC) was analyzed using immunofluorescence staining. When ADSCs and VECs were cocultured, the absorbance of cells gradually increased, reaching a peak on day 12. The highest absorbance was seen in a coculture system with a ratio of ADSCs and VECs of 1:1. The secretion of ALP and OC gradually increased in these cells and was significantly higher than controls (P < 0.01). Coculturing of ADSCs and VECs at a 1:1 ratio gave the highest secretion of ALP and OC at every time point, and was significantly higher than other groups (P < 0.01). Our results indicated that ADSCs can be induced to osteogenic differentiation by VECs in vitro, suggesting a coculture system of VECs and ADSC as a novel source of cells for bone engineering.

[Experimental study of subcutaneous osteogenesis by bone marrow stromal cells with A-PCPC in Beagle dogs].

PURPOSE: To study the osteogenic capability of the construct combined dog's bone stromal cells with active porous calcium phosphate cement (A-PCPC) in nude mice in vivo. METHODS: Isolated bone marrow stromal cells (BMSCs) were expanded and osteogenically induced in vitro. Their osteogenic phenotype was evaluated by cytochemistry. The tissue engineering complex was constructed with BMSCs/A-PCPC in vitro. After SEM scanning, the complex of BMSCs/A-PCPC was implanted into the subcutaneous tissue of the nude mice as experimental group, and A-PCPC as control group. The engineered bone was harvested 2,4,8weeks post-implantation and processed for HE staining, then evaluated by histology and histomorphometry. RESULTS: Cytochemistry showed alkaline phosphate activity, Von Kossa staining proved the formation of mineralization nodules. Scanning electron microscopy showed the cells adhered to the inner surface of the A-PCPC. HE staining showed a small group of woven bone formation 2 weeks later in the experimental group, while the formation of bone less in the control group. Woven bone turned into trabecular bone gradually at 4 weeks in the experimental group, while the control group showed a large number of bone-like tissue. Histomorphometry showed more mature bone in the experimental group than the control group at 8 weeks. CONCLUSIONS: The A-PCPC/BMSCs composites show good osteogenetic activity and could promote mineralization of the immature bone. It can be used as the bone tissue engineering scaffolds. Supported by Innovation Fund for Science and Technology Development of Pudong New District(Grant No. PKJ2009-Y19).

A follow-up study of arthroscopic combined reconstruction of anterior and posterior cruciate ligaments with allograft patellar tendon.

OBJECTIVE: To evaluate the therapeutic effect of combined reconstruction of anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) simultaneously by using allograft patellar tendon under arthroscopy. METHODS: From May 2003 to November 2005, 10 cases of ruptured ACL and PCL were fixated with compressed screws and reconstructed under arthroscopy with allograft patellar tendon simultaneously. The clinical results were evaluated according to IKDC, Lysholm, and Tegner clinical rating scales. RESULTS: All patients were followed up for 12-30 months (mean: 18 months). At the last follow-up, there was no knee extension limitation and knee flexion was between 120 degree and 135 degree,with an average of 128.38 degree. The Lysholm score of the 10 cases was 66.5+/-5.6 before operation and 89.8+/-3.4 at last follow up. The difference was statistically significant (P less than 0.01). The average Tegner activity score decreased from 6.9+/-1.7 (range: 4-9) before injury to 5.5+/-1.6 (rang:2-9) at the follow-up (P equal to 0.53). At the end of follow-up, IKDC score was graded as A in 4 cases (40.0%), B in 5 (50.0%), and C in 1 (10.0%). Of the 10 patients, 8 returned to the same sports level as before injury and 2 were under the level. CONCLUSION: Arthroscopic combined reconstruction of ACL and PCL with allograft patellar tendon has the advantages of minimal trauma in surgery and reliable satisfactory outcome.

[Bone-patellar tendon-bone graft in anterior cruciate ligament reconstruction: allograft versus autograft].

OBJECTIVE: To evaluate the clinical result of arthroscopic anterior cruciate ligament (ACL) reconstruction with either allo- or auto- bone-patellar tendon-bone (B-PT-B) grafts. METHODS: From February 2002 to January 2006, 142 of 187 cases of ACL ruptures who received ACL reconstruction with B-PT-B grafts were studied retrospectively. There were 93 male and 49 female whose age was from 15 to 57 years (mean 26 years). Patients were divided into 2 groups by graft selection: 38 patients with autograft and 104 with allograft. Clinical results were evaluated according to IKDC, Lysholm, Irgang and Larson scales. Sub-items of scales such as pain, swelling and laxity were specifically evaluated. RESULTS: All of the patients were followed up with an average of 24 months (range from 6 to 43 months). All grafts were radiographically in good position at the time of follow-up. KT-1000 examination of affected knee showed less than 3 mm anterior translation difference compared with contralateral one's. Allograft group: 85 patients got normal IKDC score (81.7%). Lysholm score 82.8 +/- 8.5, Irgang score 79.2 +/- 7.3, Larson score 86.7 +/- 3.1. Autograft group: 29 patients got normal IKDC score (76.3%). Lysholm score 84.6 +/- 9.5, Irgang score 79.5 +/- 7.6, Larson score 88.9 +/- 6.8. No significant statistical difference was found between the 2 groups in the comprehend scale (P>0.05). CONCLUSIONS: Both autograft and allograft group achieve good results, and the allograft B-PT-B could provide the similar clinical result as autograft, the preliminary result of allograft reconstruction might indicate predictable result in the future.