The alteration of the structure and macroscopic mechanical response of porcine patellar tendon by elastase digestion.

Background: The treatment of patellar tendon injury has always been an unsolved problem, and mechanical characterization is very important for its repair and reconstruction. Elastin is a contributor to mechanics, but it is not clear how it affects the elasticity, viscoelastic properties, and structure of patellar tendon. Methods: The patellar tendons from six fresh adult experimental pigs were used in this study and they were made into 77 samples. The patellar tendon was specifically degraded by elastase, and the regional mechanical response and structural changes were investigated by: (1) Based on the previous study of elastase treatment conditions, the biochemical quantification of collagen, glycosaminoglycan and total protein was carried out; (2) The patellar tendon was divided into the proximal, central, and distal regions, and then the axial tensile test and stress relaxation test were performed before and after phosphate-buffered saline (PBS) or elastase treatment; (3) The dynamic constitutive model was established by the obtained mechanical data; (4) The structural relationship between elastin and collagen fibers was analyzed by two-photon microscopy and histology. Results: There was no statistical difference in mechanics between patellar tendon regions. Compared with those before elastase treatment, the low tensile modulus decreased by 75%-80%, the high tensile modulus decreased by 38%-47%, and the transition strain was prolonged after treatment. For viscoelastic behavior, the stress relaxation increased, the initial slope increased by 55%, the saturation slope increased by 44%, and the transition time increased by 25% after enzyme treatment. Elastin degradation made the collagen fibers of patellar tendon become disordered and looser, and the fiber wavelength increased significantly. Conclusion: The results of this study show that elastin plays an important role in the mechanical properties and fiber structure stability of patellar tendon, which supplements the structure-function relationship information of patellar tendon. The established constitutive model is of great significance to the prediction, repair and replacement of patellar tendon injury. In addition, human patellar tendon has a higher elastin content, so the results of this study can provide supporting information on the natural properties of tendon elastin degradation and guide the development of artificial patellar tendon biomaterials.

M2 macrophage-derived exosomal circTMCO3 acts through miR-515-5p and ITGA8 to enhance malignancy in ovarian cancer.

Tumor-associated macrophages of the M2 phenotype promote cancer initiation and progression. Importantly, M2 macrophage-derived exosomes play key roles in the malignancy of cancer cells. Here, we report that circTMCO3 is upregulated in ovarian cancer patients, and its high expression indicates poor survival. M2-derived exosomes promote proliferation, migration, and invasion in ovarian cancer, but these effects are abolished by knockdown of circTMCO3. Furthermore, circTMCO3 functions as a competing endogenous RNA for miR-515-5p to reduce its abundance, thus upregulating ITGA8 in ovarian cancer. miR-515-5p inhibits ovarian cancer malignancy via directly downregulating ITGA8. The decreased oncogenic activity of circTMCO3-silencing exosomes is reversed by miR-515-5p knockdown or ITGA8 overexpression. Exosomal circTMCO3 promotes ovarian cancer progression in nude mice. Thus, M2 macrophage-derived exosomes promote malignancy by delivering circTMCO3 and targeting the miR-515-5p/ITGA8 axis in ovarian cancer. Our findings not only provide mechanistic insights into ovarian cancer progression, but also suggest potential therapeutic targets.

Electrospun Aligned Nanofiber Yarns Constructed Biomimetic M-Type Interface Integrated into Precise Co-Culture System as Muscle-Tendon Junction-on-a-Chip for Drug Development.

The incorporation of engineered muscle-tendon junction (MTJ) with organ-on-a-chip technology provides promising in vitro models for the understanding of cell-cell interaction at the interface between muscle and tendon tissues. However, developing engineered MTJ tissue with biomimetic anatomical interface structure remains challenging, and the precise co-culture of engineered interface tissue is further regarded as a remarkable obstacle. Herein, an interwoven waving approach is presented to develop engineered MTJ tissue with a biomimetic "M-type" interface structure, and further integrated into a precise co-culture microfluidic device for functional MTJ-on-a-chip fabrication. These multiscale MTJ scaffolds based on electrospun nanofiber yarns enabled 3D cellular alignment and differentiation, and the "M-type" structure led to cellular organization and interaction at the interface zone. Crucially, a compartmentalized co-culture system is integrated into an MTJ-on-a-chip device for the precise co-culture of muscle and tendon zones using their medium at the same time. Such an MTJ-on-a-chip device is further served for drug-associated MTJ toxic or protective efficacy investigations. These results highlight that these interwoven nanofibrous scaffolds with biomimetic "M-type" interface are beneficial for engineered MTJ tissue development, and MTJ-on-a-chip with precise co-culture system indicated their promising potential as in vitro musculoskeletal models for drug development and biological mechanism studies.

Effect of magnetized water irrigation on Cd subcellular allocation and chemical forms in leaves of Festuca arundinacea during phytoremediation.

The application of an external magnetic field has been shown to improve the Cd phytoremediation efficiency of F. arundinacea by leaf harvesting. However, the influencing mechanisms of the promoting effect have not yet been revealed. This study evaluated variations in the Cd subcellular allocation and fractions in various F. arundinacea leaves, with or without magnetized water irrigation. Over 50 % of the metal were sequestered within the cell wall in all tissues under all treatments, indicating that cell wall binding was a critical detoxification pathway for Cd. After magnetized water treatment, the metal stored in the cytoplasm of roots raised from 33.1 % to 45.3 %, and the quantity of soluble Cd in plant roots enhanced from 53.4 % to 59.0 %. The findings suggested that magnetized water mobilized Cd in the roots, and thus drove it into the leaves. In addition, the proportion of Cd in the organelles, and the concentration of ethanol-extracted Cd in emerging leaves, decreased by 13.0 % and 47.1 %, respectively, after magnetized water treatment. These results explained why an external field improved the phytoextraction effect of the plant through leaf harvesting.

Biomechanical evaluation on a new type of vertebral titanium porous mini-plate and mechanical comparison between cervical open-door laminoplasty and laminectomy: a finite element analysis.

Objective: Compare the spine's stability after laminectomy (LN) and laminoplasty (LP) for two posterior surgeries. Simultaneously, design a new vertebral titanium porous mini plate (TPMP) to achieve firm fixation of the open-door vertebral LP fully. The objective is to enhance the fixation stability, effectively prevent the possibility of "re-closure," and may facilitate bone healing. Methods: TPMP was designed by incorporating a fusion body and porous structures, and a three-dimensional finite element cervical model of C2-T1 was constructed and validated. Load LN and LP finite element models, respectively, and analyze and simulate the detailed processes of the two surgeries. It was simultaneously implanting the TPMP into LP to evaluate its biomechanical properties. Results: We find that the range of motion (ROM) of C4-C5 after LN surgery was greater than that of LP implanted with different plates alone. Furthermore, flexion-extension, lateral bending, and axial rotation reflect this change. More noteworthy is that LN has a much larger ROM on C2-C3 in axial rotation. The ROM of LP implanted with two different plates is similar. There is almost no difference in facet joint stress in lateral bending. The facet joint stress of LN is smaller on C2-C3 and C4-C5, and larger more prominent on C5-C6 in the flexion-extension. Regarding intervertebral disc pressure (IDP), there is little difference between different surgeries except for the LN on C2-C3 in axial rotation. The plate displacement specificity does not significantly differ from LP with vertebral titanium mini-plate (TMP) and LP with TPMP after surgery. The stress of LP with TPMP is larger in C4-C5, C5-C6. Moreover, LP with TMP shows greater stress in the C3-C4 during flexion-extension and lateral bending. Conclusion: LP may have better postoperative stability when posterior approach surgery is used to treat CSM; at the same time, the new type of vertebral titanium mini-plate can achieve almost the same effect as the traditional titanium mini-plate after surgery for LP. In addition, it has specific potential due to the porous structure promoting bone fusion.

S2 subunit plays a critical role in pathogenesis of TW-like avian coronavirus infectious bronchitis virus.

To investigate the critical role of the S gene in determining pathogenesis of TW-like avian infectious bronchitis virus (IBV), we generated two recombinant IBVs (rGDaGD-S1 and rGDaGD-S2) by replacing either the S1 or S2 region of GD strain with the corresponding regions from an attenuated vaccine candidate aGD strain. The virulence and pathogenicity of these recombinant viruses was assessed both in vitro and in vivo. Our results indicated the mutations in the S2 region led to decreased virulence, as evidenced by reduced virus replication in embryonated chicken eggs and chicken embryonic kidney cells as well as observed clinical symptoms, gross lesions, microscopic lesions, tracheal ciliary activity, and viral distribution in SPF chickens challenged with recombinant IBVs. These findings highlight that the S2 subunit is a key determinant of TW-like IBV pathogenicity. Our study established a foundation for future investigations into the molecular mechanisms underlying IBV virulence.

Effects of magnetic field on cd subcellular distribution and chemical speciation in Noccaea caerulescens.

Implementing an external magnetic field of suitable strength has been reported to increase Cd uptake by Noccaea caerulescence. However, only a few mechanisms promoting this efficiency have been reported. A series of culture experiments was conducted to explore how Cd subcellular distribution and speciation vary within the tissue of N. caerulescens when subjected to external magnetic fields of different intensities. Without a magnetic field, over 80% of the Cd was deposited in the cell wall and cytoplasm, indicating that cell wall retention and cytoplasm isolation are significant mechanisms for the detoxification of Cd. An external magnetic field (120 mT) increased the Cd concentrations deposited in the cytoplasm and water-soluble inorganic Cd in the roots, increasing the cell wall-bound Cd and undissolved Cd phosphate in the shoots. Meanwhile, the magnetic field increased carbonic anhydrase activity in plant shoots, except at 400 mT. These results indicated that an external field can elevate the Cd decontamination capacity of N. caerulescens by changing the subcellular compartmentalization and speciation of Cd in different tissues.

Interaction effects of magnetized water irrigation and wounding stress on Cd phytoremediation effect of Arabidopsis halleri.

In this study, the phytoremediation efficiency of Arabidopsis halleri L. in response to mechanical injury were compared between those irrigated with magnetized water and those irrigated with normal water. Under normal irrigation treatment, wounding stress increased malondialdehyde (MDA) concentrations and hydrogen peroxide (H2O2) levels in A. halleri leaves significantly, by 46.7-86.1% and 39.4-77.4%, respectively, relative to those in the intact tissues. In addition, wounding stresses decreased the content of Cd in leaves by 26.8-52.2%, relative to the control, indicating that oxidative damage in plant tissues was induced by mechanical injury, rather than Cd accumulation. There were no significant differences in MDA and H2O2 between A. halleri irrigated with magnetized water and with normal water under wounding conditions; however, the activities of catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) in the leaves of plants treated with magnetized water were significantly increased by 25.1-56.7%, 47.3-183.6%, and 44.2-109.4%, respectively. Notably, under the magnetic field, the phytoremediation effect of 30% wounded A. halleri nearly returned to normal levels. We find that irrigation with magnetized water is an economical pathway to improve the tolerance of A. halleri to inevitable mechanical injury and may recover its phytoremediation effect.

Salvage radiotherapy strategy and its prognostic significance for patients with locoregional recurrent cervical cancer after radical hysterectomy: a multicenter retrospective 10-year analysis.

OBJECTIVE: We aimed to evaluate the clinical efficacy and prognostic significance of intensity-modulated radiotherapy (IMRT)-based salvage concurrent chemoradiotherapy (CCRT) for patients with locoregional recurrence cervical cancer after radical hysterectomy and evaluated two salvage radiotherapy modes-regional RT (involved-field RT combined with regional lymph nodes) and local RT (involved-field RT). METHODS: Patients were enrolled retrospectively from January 2011 to January 2022 in three medical centers. Clinical outcomes were analyzed using the Kaplan-Meier method and a Cox proportional hazards model. Propensity score (PS) matching analysis was used to compare the two RT groups. RESULTS: There were 72 patients underwent IMRT-based salvage CCRT. The 5-year overall survival and progression-free survival rates were 65.9% and 57.6%, respectively. Univariate analysis showed that patients with stump recurrence, a lower systemic inflammation response index (SIRI), only one metastatic lesion, and received regional RT had better prognosis than their counterparts. In multivariate analysis, recurrence site was the independent prognostic factor of OS, and SIRI was that of PFS. After PS matching, there were 15 patients each in the regional RT group and local RT group. The 5-year OS rate of regional RT group was better than that of local RT group (90.9 vs. 42.4, p = 0.021). However, there was no significant difference between them in terms of PFS rate (47.1 vs. 38.1, p = 0.195). CONCLUSION: Locoregional recurrent cervical cancer treated with IMRT-based salvage therapy has a good prognosis. Recurrence site and SIRI were independent prognostic factors. Regional RT may be a better option for patients with locoregional recurrent.

Comparison of biomechanical parameters of two Chinese cervical spine rotation manipulations based on motion capture and finite element analysis.

Objective: The purpose of this study was to obtain the stress-strain of the cervical spine structure during the simulated manipulation of the oblique pulling manipulation and the cervical rotation-traction manipulation in order to compare the mechanical mechanism of the two manipulations. Methods: A motion capture system was used to record the key kinematic parameters of operating the two manipulations. At the same time, a three-dimensional finite element model of the C0-T1 full healthy cervical spine was established, and the key kinematic parameters were loaded onto the finite element model in steps to analyze and simulate the detailed process of the operation of the two manipulations. Results: A detailed finite element model of the whole cervical spine including spinal nerve roots was established, and the validity of this 3D finite element model was verified. During the stepwise simulation of the two cervical spine rotation manipulations to the right, the disc (including the annulus fibrosus and nucleus pulposus) and facet joints stresses and displacements were greater in the oblique pulling manipulation group than in the cervical rotation-traction manipulation group, while the spinal cord and nerve root stresses were greater in the cervical rotation-traction manipulation group than in the oblique pulling manipulation group. The spinal cord and nerve root stresses in the cervical rotation-traction manipulation group were mainly concentrated in the C4/5 and C5/6 segments. Conclusion: The oblique pulling manipulation may be more appropriate for the treatment of cervical spondylotic radiculopathy, while cervical rotation-traction manipulation is more appropriate for the treatment of cervical spondylosis of cervical type. Clinicians should select cervical rotation manipulations for different types of cervical spondylosis according to the patient's symptoms and needs.

In silico biomechanical analysis of poller screw-assisted small-diameter intramedullary nail in the treatment of distal tibial fractures.

Objective: To evaluate the biomechanical effects of Poller screws (PS) combined with small-diameter intramedullary nails in the treatment of distal tibial fractures at different locations and on different planes. Methods: Nine finite element (FE) models were used to simulate the placement of the intramedullary nail (IMN) and the PS for distal tibial fractures. Structural stiffness and interfragmentary motion (IFM) through the fracture were investigated to assess the biomechanical effects of the PS. The allowable stress method was used to evaluate the safety of the construct. Results: With the axial load of 500 N, the mean axial stiffness of IMN group was 973.38 ± 95.65 N/mm, which was smaller than that at positions A and B of the coronal group and sagittal group (p < 0.05). The shear IFM of the IMN group was 2.10 ± 0.02 mm, which were smaller than that at positions A and B of the coronal group and sagittal group (p < 0.05). Under physiological load, the stresses of all internal fixation devices and the nail-bone interface were within a safe range. Conclusion: In the treatment of distal tibial fractures, placing the PS in the proximal fracture block can obtain better biomechanical performance. The IMN fixation system can obtain higher structural stiffness and reduce the IFM of the fracture end by adding PS.

Plasma Polishing as a New Polishing Option to Reduce the Surface Roughness of Porous Titanium Alloy for 3D Printing.

Porous titanium alloy implants with simulated trabecular bone fabricated by 3D printing technology have broad prospects. However, due to the fact that some powder adheres to the surface of the workpiece during the manufacturing process, the surface roughness in direct printing pieces is relatively high. At the same time, since the internal pores of the porous structure cannot be polished by conventional mechanical polishing, an alternative method needs to be found. As a surface technology, plasma polishing technology is especially suitable for parts with complex shapes that are difficult to polish mechanically. It can effectively remove particles and fine splash residues attached to the surface of 3D printed porous titanium alloy workpieces. Therefore, it can reduce surface roughness. Firstly, titanium alloy powder is used to print the porous structure of the simulated trabecular bone with a metal 3D printer. After printing, heat treatment, removal of the supporting structure, and ultrasonic cleaning is carried out. Then, plasma polishing is performed, consisting of adding a polishing electrolyte with the pH set to 5.7, preheating the machine to 101.6 °C, fixing the workpiece on the polishing fixture, and setting the voltage (313 V), current (59 A), and polishing time (3 min). After polishing, the surface of the porous titanium alloy workpiece is analyzed by a confocal microscope, and the surface roughness is measured. Scanning electron microscopy is used to characterize the surface condition of porous titanium. The results show that the surface roughness of the whole porous titanium alloy workpiece changed from Ra (average roughness) = 126.9 µm to Ra = 56.28 µm, and the surface roughness of the trabecular structure changed from Ra = 42.61 µm to Ra = 26.25 µm. Meanwhile, semi-molten powders and ablative oxide layers are removed, and surface quality is improved.

Fully Biodegradable and Long-Term Operational Primary Zinc Batteries as Power Sources for Electronic Medicine.

Given the advantages of high energy density and easy deployment, biodegradable primary battery systems remain as a promising power source to achieve bioresorbable electronic medicine, eliminating secondary surgeries for device retrieval. However, currently available biobatteries are constrained by operational lifetime, biocompatibility, and biodegradability, limiting potential therapeutic outcomes as temporary implants. Herein, we propose a fully biodegradable primary zinc-molybdenum (Zn-Mo) battery with a prolonged functional lifetime of up to 19 days and desirable energy capacity and output voltage compared with reported primary Zn biobatteries. The Zn-Mo battery system is shown to have excellent biocompatibility and biodegradability and can significantly promote Schwann cell proliferation and the axonal growth of dorsal root ganglia. The biodegradable battery module with 4 Zn-Mo cells in series using gelatin electrolyte accomplishes electrochemical generation of signaling molecules (nitric oxide, NO) that can modulate the behavior of the cellular network, with efficacy comparable with that of conventional power sources. This work sheds light on materials strategies and fabrication schemes to develop high-performance biodegradable primary batteries to achieve a fully bioresorbable electronic platform for innovative medical treatments that could be beneficial for health care.

Macrolactin R from Bacillus siamensis and its antifungal activity against Botrytis cinerea.

Botrytis cinerea is listed among the most important fungal pathogens infecting strawberries. The use of biological control agents, such as Bacillus species, offers an alternative and effective way to reduce airborne pathogens. The aim of this research was to select the macrolactin R produced by Bacillus siamensis with potential for using as biological agents against the pathogenetic fungi (Botrytis cinerea) of strawberries, and to assess the mechanisms involved. Macrolactin R had significant inhibitory effects on spore germination, germ tube elongation, and mycelial growth of Botrytis cinerea. The MICs of macrolactin R inhibitions in vitro was 12.5 mg/L and The EC50 value of NJ08-3 to Botrytis cinerea spores and mycelial was 1.93 and 2.88 mg/L, respectively. Macrolactin R impacted the membrane structure of Botrytis cinerea, resulting in changes in membrane permeability and leakage of proteins and nucleic acids, then cell death. The application of the macrolactin R of Bacillus siamensis reduced the disease severity index of gray mold on strawberries. This study demonstrated that the production of macrolactin R produced by Bacillus siamensis are involved in the antifungal activity against Botrytis cinerea.

Palmitic acid induces nDNA release to cytosol and promotes microglial M1 polarization via cGAS-STING signaling pathway.

Palmitic acid (PA), the most common statured fatty acid in diets, is involved in peripheral as well as central inflammation. The M1 polarization of microglia plays an important role in PA-induced neuroinflammation. However, it is still unclear on the key factor and molecule mechanism of microglial polarization among it. Thus, we investigated whether the release of self-DNA into the cytoplasm of microglia was a consequence of PA treatment, as in aortic endothelial cells and adipocytes. RT-qPCR and immunofluorescence were performed to detect the status of cytosolic DNA and microglial polarization after PA treatment. We found that the content of cytosolic nDNA rather than mtDNA increased after PA treatment and the M1 polarization of microglia was associated with this. Moreover, the knockdown of cGAS in BV2 microglial cells demonstrated that the cGAS-STING pathway is involved in polarization process. Our results revealed that nDNA and cGAS-STING pathway are critically involved in PA-induced microglial M1 polarization. This mechanism may pose a new insight on targeting microglia may be a promising way to mitigate diet-induced early neuroinflammation.

Optimal design and biomechanical analysis of sandwich composite metal locking screws for far cortical locking constructs.

In the interests of more flexible and less stiff bridge constructs to stimulate bone healing, the technique of far cortical locking has been designed to improve locked plating constructs in terms of stress concentration, stress shielding, and inhibition of issues around fracture healing. However, far cortical locking screws currently lack objective designs and anti-fatigue designs. This study investigates an optimization algorithm to form a special locking screw composed of various metals, which can theoretically achieve the maintenance of the excellent mechanical properties of far cortical locking constructs in terms of fracture internal fixation, while maintaining the biomechanical safety and fatigue resistance of the structure. The numerical results of our study indicate that the maximum von Mises stress of the optimized construct is less than the allowable stress of the material under each working condition while still achieving sufficient parallel interfragmentary motion. Numerical analysis of high cycle fatigue indicates that the optimized construct increases the safety factor to five. A high cycle fatigue test and defect analysis indicates that the sandwich locking constructs have better fatigue resistance. We conclude that the sandwich locking construct theoretically maintains its biomechanical safety and fatigue resistance while also maintaining excellent mechanical properties for fracture internal fixation.

LncRNA LAMTOR5-AS1 sponges miR-210-3p and regulates cervical cancer progression.

AIM: Cervical cancer has attracted increasing attention in recent years, and the incidence has shown a trend of younger age. Therefore, it is an effective method to regulate the progression of cervical cancer through new prognostic biomarkers. The purpose of this study was to evaluate the potential of lncRNA LAMTOR5-AS1 (LAMTOR5-AS1) as a prognostic biomarker and reveal its regulatory role in cervical cancer. METHODS: A total of 120 patients with cervical cancer were selected as research subjects to verify the prognostic effect of LAMTOR5-AS1 in a series of experiments. The expression of LAMTOR5-AS1 in cervical cancer tissues and cells was determined by polymerase chain reaction assay. The proliferation, migration, and invasion ability of cervical cancer cells were evaluated by Cell Counting Kit-8 (CCK-8) and Transwell assay. Luciferase reporter gene detection was used to determine the mechanism of LAMTOR5-AS1 targeting miR-210-3p, and to reflect the prognostic value of LAMTOR5-AS1 according to statistical methods. RESULTS: LAMTOR5-AS1 decreased in cervical cancer tissues, while miR-210-3p expression increased. In the study of cervical cancer cells, it was found that the LAMTOR5-AS1 sponge miR-210-3p was associated with the malignant progression of cervical cancer. Overexpression of LAMTOR5-AS1 could effectively inhibit the development of cervical cancer cells and might be chosen as a prognostic biomarker of cervical cancer. CONCLUSIONS: LAMTOR5-AS1 sponges miR-210-3p and modulates the progression of cervical cancer, which predict the prognosis of cervical cancer patients.

A flexible and highly sensitive organic electrochemical transistor-based biosensor for continuous and wireless nitric oxide detection.

As nitric oxide (NO) plays significant roles in a variety of physiological processes, the capability for real-time and accurate detection of NO in live organisms is in great demand. Traditional assessments of NO rely on indirect colorimetric techniques or electrochemical sensors that often comprise rigid constituent materials and can hardly satisfy sensitivity and spatial resolution simultaneously. Here, we report a flexible and highly sensitive biosensor based on organic electrochemical transistors (OECTs) capable of continuous and wireless detection of NO in biological systems. By modifying the geometry of the active channel and the gate electrodes of OECTs, devices achieve optimum signal amplification of NO. The sensor exhibits a low response limit, a wide linear range, high sensitivity, and excellent selectivity, with a miniaturized active sensing region compared with a conventional electrochemical sensor. The device demonstrates continuous detection of the nanomolar range of NO in cultured cells for hours without significant signal drift. Real-time and wireless measurement of NO is accomplished for 8 d in the articular cavity of New Zealand White rabbits with anterior cruciate ligament (ACL) rupture injuries. The observed high level of NO is associated with the onset of osteoarthritis (OA) at the later stage. The proposed device platform could provide critical information for the early diagnosis of chronic diseases and timely medical intervention to optimize therapeutic efficacy.

[Analysis of hereditary coagulation factor â ª deficiency in a Chinese pedigree with compound heterozygous mutations].

OBJECTIVE: To explore the molecular mechanisms of a Chinese pedigree with hereditary factor Ⅺ (FⅪ) deficiency. METHODS: All of the 15 exons, flanking sequences of the FⅪ gene and the corresponding mutation sites of family members were analyzed by the Sanger sequencing, followed by the extraction of the peripheral blood genomic DNA. And all the results were verified by the reverse sequencing. The conservation of the mutated sites was analyzed by the ClustalX-2.1-win. Three online bioinformatics software tools, including Mutation Taster, PolyPhen2 and the PROVEAN, were used to assess the possible impact of the mutations. Swiss-pdbviewer software was used to analyze the effects of mutant amino acids on protein structure. RESULTS: Genetic analysis revealed that the proband had compound heterozygous mutations including a nonsense mutation of c.1107C>A (Tyr369stop) in exon 10 and missense mutation of c.1562A>G (Tyr521Cys) in exon 13. The same c.1107C>A (Tyr369stop) was present in her father, the same c.1562A>G (Tyr521Cys) was present in both her mother and daughter. Conservation analysis indicated that Tyr521 was a highly conserved site during evolution. The prediction of pathogenicity showed that both c.1107C>A and c.1562A>G were pathogenic mutations. Protein structure prediction showed that in the wild type FⅪ protein structure, Tyr521 formed a hydrogen bond with the Lys572 and Ile388, respectively. When Tyr521 was replaced by Cys521, the original benzene ring structure disappeared, and side chains of Lys572 added a hydrogen bond with the Cys521, which may change protein catalytic domain structure. When Tyr369 was mutated to a stop codon, resulting in the truncated protein. CONCLUSION: The compound heterozygous mutations including the c.1107C>A heterozygous missense variant in exon 10 and the c.1562A>G heterozygous nonsense mutation in exon 13 may be responsible for the hereditary factor Ⅺ deficiency in this Chinese pedigree.

Patterns and driving forces of the agricultural water footprint of Chinese cities.

The patterns and determinants of different types of agricultural water footprints in China are poorly understood at the prefecture-city level. In this paper, we evaluate Chinese agricultural water footprints from 2000 to 2017 and analyzed their spatio-temporal characteristics. Our estimation results show that the annual average agricultural water footprint in China was 5.038 × 109 m3, and the proportions of green water, blue water, and gray water were 70%, 9%, and 21%, respectively. In addition, high agricultural water-footprint cities with obvious urban agglomeration effects are mainly located in the Northeast, the Huanghuai River, the Yangtze River Basin, and Northwestern of Xinjiang, while low agricultural water-footprint cities are concentrated in high coastal urbanization-level areas or less developed agricultural areas of the west. We also investigate their determinants using a spatio-temporal fixed-effect model and find that GDP per capita, total investment in fixed assets, the income level of rural residents, the proportion of food grown, spray and drip irrigation technology, low-pressure pipe irrigation technology and seepage control irrigation technology have significant positive impacts on the agricultural water footprint. In contrast, the proportion of secondary and tertiary industries, social retail consumption, urbanization, technology expenditure, and the effective irrigation area proportion have a significant inhibitory effect. The primary determinants of the agricultural water footprint also vary substantially across water footprint categories (green, blue, and gray water footprints) and regions. Our findings imply that the agricultural water footprint should be incorporated into city water resource management and monitoring system.