Optimizing conditioning regimen with low-dose irradiation or busulfan enables the outcome of transplantation from a 6-7/8 HLA-matched donor comparable to that from an 8/8 HLA-matched unrelated donor in severe aplastic anemia patients under 40 years.

With the dramatic improvements in outcomes following alternative donor hematopoietic stem cell transplantation (HSCT), interest in the use of alternative donors in severe aplastic anemia (SAA) is increasing. We conducted a multicenter prospective study to explore the efficiency and safety of upfront HSCT from a 6-8/8 HLA-matched unrelated donor (MUD) or 6-7/8 HLA-matched related donor (MRD) in acquired SAA patients under 40 years. Between August 2014 and July 2017, 115 patients were enrolled, including 48 (41.7%) patients receiving grafts from an 8/8 MUD, 25 (21.7%) from a 6-7/8 MRD, and 42 (36.5%) from a 6-7/8 MUD. The incidence of grade II-IV acute graft-versus-host disease (GVHD) was higher in the 6-7/8 MUD group than in the 8/8 MUD group (42.9% vs. 12.8%, P=0.001). The corresponding incidence in the 6-7/8 MRD group was comparable to that in the 8/8 MUD group (21.7% vs. 12.8%, P=0.332). There was no significant difference in the incidence of chronic GVHD (24.3%, 13.6%, and 17.9%, P=0.676), graft failure (2.4%, 8.0%, and 6.3%, P=0.551), overall survival (85.7%, 96.0%, and 87.5%, P=0.424), and failure-free survival (83.3%, 88.0%, and 83.3%, P=0.885) among the three groups (6-7/8 MUD, 6-7/8 MRD, and 8/8 MUD). In multivariate analysis, conditioning regimen without low-dose irradiation or busulfan was associated with an inferior failure-free survival (HR=2.973, P=0.042). In conclusion, after an intensified conditioning regimen with additional low-dose irradiation or busulfan, the outcome of HSCT from a 6-7/8 MRD or 6-7/8 MUD is comparable to that from an 8/8 MUD.

Mapping the Endothelial Cell S-Sulfhydrome Highlights the Crucial Role of Integrin Sulfhydration in Vascular Function.

BACKGROUND: In vascular endothelial cells, cysteine metabolism by the cystathionine γ lyase (CSE), generates hydrogen sulfide-related sulfane sulfur compounds (H2Sn), that exert their biological actions via cysteine S-sulfhydration of target proteins. This study set out to map the "S-sulfhydrome" (ie, the spectrum of proteins targeted by H2Sn) in human endothelial cells. METHODS: Liquid chromatography with tandem mass spectrometry was used to identify S-sulfhydrated cysteines in endothelial cell proteins and ß3 integrin intraprotein disulfide bond rearrangement. Functional studies included endothelial cell adhesion, shear stress-induced cell alignment, blood pressure measurements, and flow-induced vasodilatation in endothelial cell-specific CSE knockout mice and in a small collective of patients with endothelial dysfunction. RESULTS: Three paired sample sets were compared: (1) native human endothelial cells isolated from plaque-free mesenteric arteries (CSE activity high) and plaque-containing carotid arteries (CSE activity low); (2) cultured human endothelial cells kept under static conditions or exposed to fluid shear stress to decrease CSE expression; and (3) cultured endothelial cells exposed to shear stress to decrease CSE expression and treated with solvent or the slow-releasing H2Sn donor, SG1002. The endothelial cell "S-sulfhydrome" consisted of 3446 individual cysteine residues in 1591 proteins. The most altered family of proteins were the integrins and focusing on ß3 integrin in detail we found that S-sulfhydration affected intraprotein disulfide bond formation and was required for the maintenance of an extended-open conformation of the ß leg. ß3 integrin S-sulfhydration was required for endothelial cell mechanotransduction in vitro as well as flow-induced dilatation in murine mesenteric arteries. In cultured cells, the loss of S-sulfhydration impaired interactions between ß3 integrin and Gα13 (guanine nucleotide-binding protein subunit α 13), resulting in the constitutive activation of RhoA (ras homolog family member A) and impaired flow-induced endothelial cell realignment. In humans with atherosclerosis, endothelial function correlated with low H2Sn generation, impaired flow-induced dilatation, and failure to detect ß3 integrin S-sulfhydration, all of which were rescued after the administration of an H2Sn supplement. CONCLUSIONS: Vascular disease is associated with marked changes in the S-sulfhydration of endothelial cell proteins involved in mediating responses to flow. Short-term H2Sn supplementation improved vascular reactivity in humans highlighting the potential of interfering with this pathway to treat vascular disease.

Frequency-following response among neonates with progressive moderate hyperbilirubinemia.

OBJECTIVE: To evaluate the feasibility of auditory monitoring of neurophysiological status using frequency-following response (FFR) in neonates with progressive moderate hyperbilirubinemia, measured by transcutaneous (TcB) levels. STUDY DESIGN: ABR and FFR measures were compared and correlated with TcB levels across three groups. Group I was a healthy cohort (n = 13). Group II (n = 28) consisted of neonates with progressive, moderate hyperbilirubinemia and Group III consisted of the same neonates, post physician-ordered phototherapy. RESULT: FFR amplitudes in Group I controls (TcB = 83.1 ± 32.5µmol/L; 4.9 ± 1.9 mg/dL) were greater than Group II (TcB = 209.3 ± 48.0µmol/L; 12.1 ± 2.8 mg/dL). After TcB was lowered by phototherapy, FFR amplitudes in Group III were similar to controls. Lower TcB levels correlated with larger FFR amplitudes (r = -0.291, p = 0.015), but not with ABR wave amplitude or latencies. CONCLUSION: The FFR is a promising measure of the dynamic neurophysiological status in neonates, and may be useful in tracking neurotoxicity in infants with hyperbilirubinemia.

Soluble epoxide hydrolase promotes astrocyte survival in retinopathy of prematurity.

Polyunsaturated fatty acids such as docosahexaenoic acid (DHA) positively affect the outcome of retinopathy of prematurity (ROP). Given that DHA metabolism by cytochrome P450 and soluble epoxide hydrolase (sEH) enzymes affects retinal angiogenesis and vascular stability, we investigated the role of sEH in a mouse model of ROP. In WT mice, hyperoxia elicited tyrosine nitration and inhibition of sEH and decreased generation of the DHA-derived diol 19,20-dihydroxydocosapentaenoic acid (19,20-DHDP). Correspondingly, in a murine model of ROP, sEH-/- mice developed a larger central avascular zone and peripheral pathological vascular tuft formation than did their WT littermates. Astrocytes were the cells most affected by sEH deletion, and hyperoxia increased astrocyte apoptosis. In rescue experiments, 19,20-DHDP prevented astrocyte loss by targeting the mitochondrial membrane to prevent the hyperoxia-induced dissociation of presenilin-1 and presenilin-1-associated protein to attenuate poly ADP-ribose polymerase activation and mitochondrial DNA damage. Therapeutic intravitreal administration of 19,20-DHDP not only suppressed astrocyte loss, but also reduced pathological vascular tuft formation in sEH-/- mice. Our data indicate that sEH activity is required for mitochondrial integrity and retinal astrocyte survival in ROP. Moreover, 19,20-DHDP may be more effective than DHA as a nutritional supplement for preventing retinopathy in preterm infants.

The activated iron system for phosphorus recovery in aqueous environments.

Finding a good sorbent for phosphorus (P) recovery from the aquatic environment is critical for preventing eutrophication and providing P resources. The activated iron system (mainly consisted of zero-valent iron (ZVI), Fe3O4 and Fe2+) has been reported to exhibit a favorable performance towards various contaminants in wastewater, but its effect on P recovery has not been studied systematically. In this study, we used Fe2+-nitrate pretreatment reaction to prepare the activated iron system and then applied it to P recovery. Results show that more than 99% P was removed from water in 60 min; co-existing anions (NO3-, Cl- and SO42-) and natural organic matter (NOM) had little effect on P removal. The P removal capacity of activated iron system is very high compared with currently reported sorbents. Externally-supplied Fe2+ plays an important role on P removal in the system. Regeneration study shows that the activated iron system exhibited stable P recovery ability by using 0.1 M NaOH solution. Various methods were applied to characterize the ZVI and iron corrosion, and results conclude that sorption precipitation, and co-precipitation contribute to P removal. This method will be promising and have an application potential in the field for efficient and cost-effective recovery of P with cheap microscale zero valent iron.

Evaluation of two algorithms for detecting human frequency-following responses to voice pitch.

OBJECTIVE: Voice pitch carries important cues for speech perception in humans. Recent studies have shown the feasibility of recording the frequency-following response (FFR) to voice pitch in normal-hearing listeners. The presence of such a response, however, has been dependent on subjective interpretation of experimenters. The purpose of this study was to develop and test an automated procedure including a control-experimental protocol and response-threshold criteria suitable for extracting FFRs to voice pitch, and compare the results to human judgments. DESIGN: A set of four Mandarin tones (Tone 1 flat; Tone 2 rising; Tone 3 dipping; and Tone 4 falling) were prepared to reflect the four contrastive pitch contours. Two distinctive algorithms, short-term autocorrelation in the time domain and narrow-band spectrogram in the frequency domain, were used to estimate the Frequency Error, Slope Error, Tracking Accuracy, Pitch Strength and Pitch-Noise Ratio of the recordings from individual listeners as well as the power and false-positive rates of each algorithm. STUDY SAMPLE: Eleven native speakers (five males; age: mean ± SD = 31.4 ± 4.7 years) of Mandarin Chinese were recruited. RESULTS: The results demonstrated that both algorithms were suitable for extracting FFRs and the objective measures showed comparable results to human judgments. CONCLUSIONS: The automated procedure used in this study, including the use of the control-experimental protocol and response thresholds used for each of the five objective indices, can be used for difficult-to-test patients and may prove to be useful as an assessment and diagnostic method in both clinical and basic research efforts.

[Effects of manipulation therapy in repairing rabbit articular cartilage defects].

OBJECTIVE: To evaluate the efficacy of manipulation therapy in repairing thin-layer and thick-layer articular cartilage defects in rabbits. METHODS: Twenty-four rabbits were randomly divided into manipulation group and control group. Chondral defects in the center of condyles medialis femoris (punch diameter 2.0 mm, thin-layer defect, without damaging the subchondral bone) and the center of facies patellar femoris (punch diameter 3.0 mm, thick-layer defect, without damaging the subchondral bone) were made in both side knee joints of the rabbits. Four weeks after chondral defect, the rabbits in the manipulation group were treated with manipulation therapy, while the rabbits in the control group didn't take any therapeutic measure. The rabbits were sacrificed after 4-, 8- and 12-week manipulation treatment to obtain the distal femur. The appearance of the chondral defect was first assessed, and then regenerated tissues were stained respectively with hematoxylin and eosin (HE), safranin O and immunohistochemical methods to observe the changes in pathology. The regenerated tissue O'Driscoll score system was also used to evaluate the efficacy of manipulation therapy. RESULTS: After manipulation treatment, although the thin-layer articular cartilage defects of the control group were partly repaired, a sharp gap between the regenerated tissue and the normal cartilage was visible. The range of the thick-layer articular cartilage defect in the control group became smaller, and the regenerated tissue only consisted of fiber tissue. On the contrary, both the thin-layer defects and the thick-layer defects in the manipulation group were well repaired. The appearance of the regenerated tissues was close to the normal cartilage. The gap between regenerated tissue and normal tissue was disappeared and the regenerated tissue was well integrated with around tissues. HE staining showed that the regenerated tissues in the manipulation group were similar to hyaline cartilage. Safranin O staining showed that matrix stain of the regenerated tissues in the manipulation group was similar to the normal cartilage, but no stain was observed in the control group. The type II collagen immunohistochemical staining showed that the color of the regenerated tissues was a little darker than that of the normal cartilage in the two kinds of defects in the manipulation group, and the two kinds of defects in the control group were little stained. The O'Driscoll cartilage scores of the two kinds of defects in the manipulation group were higher than those in the control group at different time points (P<0.01), but there were no differences in O'Driscoll scores in the manipulation group among the different time points. CONCLUSION: Manipulation therapy is effective in repairing the thick-layer and thin-layer articular cartilage defects. The regenerated tissue is constructed with hyaline cartilage.

[Application of magnetic resonance imaging in evaluating the effects of manipulation on knee osteoarthritis].

OBJECTIVE: To observe and evaluate the effects of manipulation on knee osteoarthritis (KOA) using T2-mapping and magnetic resonance imaging (MRI)-based volume measurements. METHODS: Forty-five cases of KOA were involved retrospectively in the study, and the patients were composed of 9 males and 36 females with the mean age of (57.4+/-6.1) years. The cases were treated with manipulative therapy once or twice per week for 12 months. MRI of each knee was performed separately by using 1.5-T MRI equipment before and during the treatment. Average cartilage depth, MRI grading of cartilage defects, cartilage volume, average T2 values in patella cartilage and femoral condyle, and bone marrow edema area were detected respectively. RESULTS: The knee joint cartilage thickness from MRI began to increase after 6-month treatment (P<0.05). The cartilage volume increase was obviously observed after 12-month treatment, and there was a significant difference (F=14.64, P<0.01). MRI grading of cartilage defects decreased from IIIA to IIB after 3-month treatment (Z=17.96, P<0.05). The average T2 value in patella cartilage decreased after 9-month treatment (F=3.11, P<0.05), but there were no differences in cartilage from tibial plateau and femoral condyle compartments after the treatment. The bone marrow edema area in femoral condyle began to diminish at 3-month treatment (t=-4.53, P<0.01), and the bone marrow edema area in cartilage patella was diminished after 6-month treatment (t=-5.53, P<0.01). CONCLUSION: T2-mapping and cartilage volume measurement are suitable for evaluating the manipulative therapy on KOA. Traditional Chinese manipulation therapy is an effective method for KOA in the cartilage recovery.

Astragaloside IV attenuates hypoxia-induced cardiomyocyte damage in rats by upregulating superoxide dismutase-1 levels.

1. Astragaloside IV (AST-IV) is purified from a natural plant product. Previous studies have shown that AST-IV has anti-oxidant activity. In the present study, we investigated the effect and mechanism of action AST-IV on rat cardiomyocytes subjected to hypoxic conditions (up to 12 h). 2. Cardiomyocytes were prepared from neonatal rats and cultured under normoxic or hypoxic conditions in the absence or presence of AST-IV (12.5, 25 or 50 microg/mL). Cell viability, malondialdehyde (MDA) levels, activity and expression of superoxide dismutase (SOD)-1 (mRNA and protein levels determined by reverse transcription-polymerase chain reaction and western blotting, respectively) and reactive oxygen species (ROS; determined by 2',7'-dichlorodihydrofluorescein diacetate) were investigated under these culture conditions. Intracellular localization of AST-IV was tested using fluorescein isothiocyanate-labelled AST-IV. 3. Hypoxic culture reduced the viability of cardiomyocytes, which was improved following treatment with 25 or 50 microg/mL AST-IV. Under hypoxic conditions, MDA levels were double those under control conditions. Astragaloside IV (25 and 50 microg/mL) dose-dependently reduced the increase in MDA seen in hypoxic cardiomyocytes. 4. Fluorescein isothiocyanate-labelled AST-IV entered cardiomyocytes and was localized mainly within the cytoplasm. 5. Under hypoxic conditions, SOD-1 activity was decreased, but mRNA and protein expression increased, compared with normoxia. Following treatment with 25 microg/mL AST-IV, SOD-1 activity and expression were increased under both normoxic and hypoxic conditions. The ROS scavenging effect of AST-IV was abolished in the presence of the SOD inhibitor sodium diethyl dithiocarbamate (25 micromol/L). 6. These in vitro results show that AST-IV protects cardiomyocytes from oxidative stress-mediated injury under hypoxic conditions. A major part of this action is achieved by upregulation of SOD-1 content and activity within the cell cytoplasm.

[Magnetic resonance assessment the effect of manipulation on knee cartilaginous recovery of osteoarthritis].

OBJECTIVE: To observe and assess the effect of manipulation on knee cartilaginous recovery with knee osteoarthritis (OA) by using magnetic resonance (MR). METHODS: Fifty cases which were suffering from knee OA involved this retrospective study. They were matched-pairs into 2 groups according to their gender, age and severity. Treated with manipulation once a week in one year for manipulation group patients, compared with those orally use with 500 mg glucosamine sulfate (GS) three times per day. Knee cartilage MR were performed before treatment and on 3, 6, 12 months after treatment, the maximum defect diameter and volume of knee cartilage were assessed with Noyes Score. RESULT: Both Noyes Score declined in the two groups. But Noyes Score of the manipulation group significantly decreased 6 months after treatment, the same tendency was observed just 12 months after treatment in another group. The maximum defect diameter of knee cartilage began to diminish at 3 months after treatment in the manipulation group, grew significantly at 6 and 12 months after treatment compared with before treatment. In the GS group, there was no significantly deference in the maximum defect diameter of knee cartilage between after and before treatment. The volume of knee cartilage in manipulation group was greater than the GS group at 3, 6, 12 months after treatment and significantly increased at 6 months after treatment and grew 58 percent 12 months after treatment. The volume of knee cartilage in GS group had no significantly change, though had a tendency to increase. CONCLUSION: Manipulation is effective to treatment of knee osteoarthritis by decreasing the maximum defect diameter and increasing the volume of knee cartilage.