Accumulation of ß-aminoisobutyric acid mediates hyperalgesia in ovariectomized mice through Mas-related G protein-coupled receptor D signaling.

Hyperalgesia is typified by reduced pain thresholds and heightened responses to painful stimuli, with a notable prevalence in menopausal women, but the underlying mechanisms are far from understood. ß-Aminoisobutyric acid (BAIBA), a product of valine and thymine catabolism, has been reported to be a novel ligand of the Mas-related G protein coupled receptor D (MrgprD), which mediates pain and hyperalgesia. Here, we established a hyperalgesia model in 8-week-old female mice through ovariectomy (OVX). A significant increase in BAIBA plasma level was observed and was associated with decline of mechanical withdrawal threshold, thermal and cold withdrawal latency in mice after 6 weeks of OVX surgery. Increased expression of MrgprD in dorsal root ganglion (DRG) was shown in OVX mice compared to Sham mice. Interestingly, chronic loading with BAIBA not only exacerbated hyperalgesia in OVX mice, but also induced hyperalgesia in gonadally intact female mice. BAIBA supplementation also upregulated the MrgprD expression in DRG of both OVX and intact female mice, and enhanced the excitability of DRG neurons in vitro. Knockout of MrgprD markedly suppressed the effects of BAIBA on hyperalgesia and excitability of DRG neurons. Collectively, our data suggest the involvement of BAIBA in the development of hyperalgesia via MrgprD-dependent pathway, and illuminate the mechanisms underlying hyperalgesia in menopausal women.

Tanshinone IIA attenuates osteoarthritis via inhibiting aberrant angiogenesis in subchondral bone.

Excessive angiogenesis in subchondral bone is a pathological feature of osteoarthritis (OA). Tanshinone IIA (TIIA), an active compound found in Salvia miltiorrhiza, demonstrates significant anti-angiogenic properties. However, the effect of TIIA on abnormal subchondral angiogenesis in OA is still unclear. This study aims to investigate the mechanism of TIIA in modulating subchondral bone angiogenesis during OA and assess its therapeutic potential in OA. Our findings demonstrate that TIIA attenuated articular cartilage degeneration, normalized subchondral bone remodeling, and effectively suppressed aberrant angiogenesis within subchondral bone in monosodium iodoacetate (MIA)-induced OA mice. Additionally, the angiogenesis capacity of primary CD31hiEmcnhi endothelial cells was observed to be significantly reduced after treatment with TIIA in vitro. Mechanically, TIIA diminished the proportion of hypertrophic chondrocytes, ultimately leading to a substantial reduction in the secretion of vascular endothelial growth factor A (VEGFA). The supernatant of hypertrophic chondrocytes promoted the tube formation of CD31hiEMCNhi endothelial cells, whereas TIIA inhibited this process. Furthermore, TIIA effectively suppressed the expression of vascular endothelial growth factor receptor 2 (VEGFR2) along with its downstream MAPK pathway in CD31hiEmcnhi endothelial cells. In conclusion, our data indicated that TIIA could effectively inhibit the abnormal angiogenesis in subchondral bone during the progression of OA by suppressing the VEGFA/VEFGR2/MAPK pathway. These findings significantly contribute to our understanding of the abnormal angiogenesis in OA and offer a promising therapeutic target for OA treatment.

Age-related decline in melatonin contributes to enhanced osteoclastogenesis via disruption of redox homeostasis.

BACKGROUND: Increased oxidative stress contributes to enhanced osteoclastogenesis and age-related bone loss. Melatonin (MT) is an endogenous antioxidant and declines with aging. However, it was unclear whether the decline of MT was involved in the enhanced osteoclastogenesis during the aging process. METHODS: The plasma level of MT, oxidative stress status, bone mass, the number of bone marrow-derived monocytes (BMMs) and its osteoclastogenesis were analyzed in young (3-month old) and old (18-month old) mice (n = 6 per group). In vitro, BMMs isolated from aged mice were treated with or without MT, followed by detecting the change of osteoclastogenesis and intracellular reactive oxygen species (ROS) level. Furthermore, old mice were treated with MT for 2 months to investigate the therapeutic effect. RESULTS: The plasma level of MT was markedly lower in aged mice compared with young mice. Age-related decline in MT was accompanied by enhanced oxidative stress, osteoclastogenic potential and bone loss. MT intervention significantly suppressed the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis, decreased intracellular ROS and enhanced antioxidant capacity of BMMs from aged mice. MT supplementation significantly attenuated oxidative stress, osteoclastogenesis, bone loss and deterioration of bone microstructure in aged mice. CONCLUSIONS: These results suggest that age-related decline of MT enhanced osteoclastogenesis via disruption of redox homeostasis. MT may serve as a key regulator in osteoclastogenesis and bone homeostasis, thereby highlighting its potential as a preventive agent for age-related bone loss.

Three-dimensional femur morphology analysis for the optimal location of subtrochanteric osteotomy with an implanted Wagner cone stem in total hip arthroplasty for Crowe type IV developmental dysplasia of the hip.

BACKGROUND: This study aimed to accurately evaluate the matching of proximal and distal femoral segments and fitting of the femur-femoral stem in patients with Crowe type IV developmental dysplasia of the hip (DDH) who have undergone subtrochanteric osteotomy at different locations with an implanted Wagner cone stem to improve the rate of the bone union at the osteotomy site. METHODS: Three-dimensional femur morphology of 40 patients with Crowe type IV DDH was evaluated at each cross-section to determine the femoral cortical bone area. This study focused on five osteotomy lengths (2.5, 3, 3.5, 4, and 4.5 cm). The overlapped area between the proximal and distal cortical bone segments was defined as the contact area (S, mm2), and the contact area to distal cortical bone area ratio was defined as the coincidence rate (R). Three indicators were used to evaluate the matching and fitting of the osteotomy sites with the implanted Wagner cone stems: (1) higher S and R between the proximal and distal segments; (2) the effective fixation length of the femoral stem at the distal segments being at least 1.5 cm; and (3) osteotomy did not involve the isthmus. RESULTS: In all groups, S significantly decreased in the two proximal levels above the 0.5 cm level below the lesser trochanter (LT) compared with those below this level. In comparison, at osteotomy lengths from 2.5 to 4 cm, R significantly decreased in the three proximal levels. The optimal osteotomy levels ranged from 1.5 and 2.5 cm below the LT for an appropriately sized stem. CONCLUSIONS: Subtrochanteric osteotomy at the optimal level not only ensures fitting of the femur-femoral stem but also meets the requirements of a higher S and R to ensure adequate reduction and stabilization at the osteotomy site, which may contribute to the bone union. Although the optimal osteotomy level varies with the size of the femoral stem and the length of the subtrochanteric osteotomy, the optimal osteotomy levels for an appropriately sized Wagner cone femoral stem implantation range from 1.5 to 2.5 cm below the LT.

CircRBM23 regulates the switch between osteogenesis and adipogenesis of mesenchymal stem cells via sponging miR-338-3p.

BACKGROUND: The disruption of the balance between osteogenic and adipogenic differentiation of mesenchymal stem cells (MSCs) in bone marrow contributes to the adipocytes accumulation and bone loss, which leads to the development of osteoporosis (OP). The circular RNA (circRNA), circRBM23, was generated from the RNA binding motif protein 23 (RBM23) gene. It was reported that circRBM23 was down-regulated in OP patients, but it remains unknown whether its down-regulation is involved in the lineage switch of MSCs. OBJECTIVE: We aimed to explore the role and mechanism of circRBM23 in regulating the switch between osteogenic and adipogenic differentiation of MSCs. METHODS: The expression and function of circRBM23 in vitro were detected by qRT-PCR, alizarin red staining, and oil Red O staining. The interactions between circRBM23 and microRNA-338-3p (miR-338-3p) were analyzed by RNA pull-down assay, FISH, and dual-luciferase reporter assay. MSCs treated with lentivirus overexpression of circRBM23 was applied for both in vitro and in vivo experiments. RESULTS: CircRBM23 was expressed at lower levels in OP patients. Besides, circRBM23 was up-regulated during osteogenesis and down-regulated during adipogenesis of MSCs. CircRBM23 could promote the osteogenic differentiation but inhibit the adipogenic differentiation of MSCs. Mechanistically, circRBM23 acted as a sponge for microRNA-338-3p (miR-338-3p) to enhance the expression of RUNX family transcription factor 2 (RUNX2). CONCLUSIONS: Our research indicates that circRBM23 could promote the switch from adipogenic to osteogenic differentiation of MSCs via sponging miR-338-3p. It might improve the understanding of the lineage switch of MSCs and provide a potential target for diagnosing and treating OP.

[Propensity score-matched study of uni-condylar arthroplasty in elderly patients with knee osteoarthritis].

OBJECTIVE: To investigate the clinical efficacy of unicompartmental knee arthroplasty (UKA) in the treatment of knee osteoarthritis in patients over 75 years old. METHODS: The clinical efficacy of primary fixed platform UKA in patients with osteoarthritis, was retrospectively analyzed from October 2014 to November 2020. Age, body mass index (BMI), range of motion (ROM), preoperative joint function score, the quality of life score and other preoperative indicators were measured by propensity score matching (PSM). The patients were divided into elderly group (≥75 years old) and control group (<75 years old). Oxford knee score(OKS), Western Ontario McMaster Universities osteoarthritis index(WOMAC), Short Form-12 including physical component summary (PCS), mental component summary(MCS), minimal clinically important difference(MCID ) and clinical complications were evaluated preoperatively and postoperatively. RESULTS: A total of 514 patients were analyzed, 428 patients fulfilled the inclusion criteria. A propensity-score matching study was conducted to eliminate confounding factors. After 1∶2 propensity match, there were 84 patients in elderly group (≥75 years), age ranged from 75 to 88 years old, with an average of (78.79±3.08) years old, and 168 patients in control group (<75 years), age ranged from 47 to 74 years old, with an average of (64.10±5.96)years old. The follow-up duration of two groups ranged from 12 to 84 months with an average of (29.35±16.52) months in elderly group, and 12 to 85 months with an average of (31.83±17.34) months in control group. There was only significant difference in age between the elderly and control groups preoperatively (P<0.01). Postoperatively, the elderly group showed significantly higher WOMAC (P<0.01) and lower SF-12 PCS scores (P<0.01) as compared to the control group. There was no significant difference between the elderly group and the control group in knee range of motion, OKS and the proportion of each scoring system reaching the minimum clinical difference value (P>0.05). In the aspect of preperative complications, the elderly group exhibited more surgical site complications and postoperative delirium compared to control group(P<0.05). The differences in other indicators including deep vein thrombosis, acute urinary retention, cardiovascular events, cerebrovascular events and radiolucent lines around prothesis were not statistically significant(P>0.05). CONCLUSION: UKA in the treatment of elderly patients over 75 years old with knee osteoarthritis was safe and feasible, and could obtain satisfactory short-term efficacy.

Induction of Anxiety-Like Phenotypes by Knockdown of Cannabinoid Type-1 Receptors in the Amygdala of Marmosets / 神经科学通报·英文版

The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases, such as depression and anxiety. Meanwhile, the endocannabinoid system plays a crucial role in regulating emotions and mainly functions through the cannabinoid type-1 receptor (CB1R), which is strongly expressed in the amygdala of non-human primates (NHPs). However, it remains largely unknown how the CB1Rs in the amygdala of NHPs regulate mental diseases. Here, we investigated the role of CB1R by knocking down the cannabinoid receptor 1 (CNR1) gene encoding CB1R in the amygdala of adult marmosets through regional delivery of AAV-SaCas9-gRNA. We found that CB1R knockdown in the amygdala induced anxiety-like behaviors, including disrupted night sleep, agitated psychomotor activity in new environments, and reduced social desire. Moreover, marmosets with CB1R-knockdown had up-regulated plasma cortisol levels. These results indicate that the knockdown of CB1Rs in the amygdala induces anxiety-like behaviors in marmosets, and this may be the mechanism underlying the regulation of anxiety by CB1Rs in the amygdala of NHPs.

[Effect of open wedge tibial high osteotomy on patella position and joint function].

OBJECTIVE: To investigate the effect of open wedge tibial high osteotomy on patella position, anterior knee pain and joint function. METHODS: From June 2016 to June 2021, 109 patients (111 knees) with medial knee osteoarthropathy treated by open wedge tibial high osteotomy were included according to the inclusion and exclusion criteria, including 41 males and 68 females;the age ranged from 38 to 78 years old with an average of(57.98±7.07) years;the course of disease ranged from 1 to 36 months with an average of (8.58±6.91) months. The femoral tibial angle(FTA), medial proximal tibial angle(MPTA), weight bearing line(WBL) percentage, Caton Deschamps index (CD index), lateral patella tilt angle (LPTA) and lateral patella shift (LPS) were observed and compared before and after operation. Lysholm score was used to evaluate the knee function, visual analogue scale(VAS) was used to evaluate the degree of anterior knee pain, and Kellgren Lawrence(K-L) grading system was used to evaluate the progress of patellofemoral osteoarthritis. RESULTS: All patients were followed up for 6 to 38 months with an average of (12.41±2.40) months. The preoperative FTA, MPTA, WBL percentage, CD index, and LPTA were significantly different from those at the last follow-up(P<0.05). There was no significant difference between before and after LPS operation(P=0.78). Lysholm score increased from (58.79±7.90) scores to (76.05±7.36) scores (P<0.05). The VAS of anterior knee pain decreased from (3.28±1.95) scores to(1.07±1.75) scores(P<0.05). Knee patellofemoral osteoarthritis showed progressive changes, but there was no significant difference in K-L grading before and after operation (P>0.05). CONCLUSION: After open wedge tibial high osteotomy, the position of patella is lowered and the patella is tilted outward, but the knee function and anterior knee pain are significantly improved. Adverse changes in patella position caused by open wedge tibial high osteotomy may not affect clinical outcomes.

Mid-Term Outcomes of Cemented Stem and Subtrochanteric Shortening Derotational Osteotomy in Total Hip Arthroplasty for Crowe IV Developmental Dysplasia.

OBJECTIVE: Performing subtrochanteric osteotomy with cemented components in patients with Crowe IV developmental dysplasia of the hip (DDH) is technically challenging and not widely reported. This study aimed to evaluate the mid-term outcomes of cemented stem total hip arthroplasty (THA) with subtrochanteric femoral shortening and transverse derotational osteotomy in patients with Crowe IV DDH. METHODS: Data collected from patients with Crowe IV DDH who underwent cemented stem THA with subtrochanteric femoral shortening and transverse derotational osteotomy between 2010 and 2018 were retrospectively evaluated. The cemented Lubinus SP II femoral component and the cementless CombiCup acetabular component were used together in all cases. These data, including Harris hip scores, limb length discrepancy (LLD), severity of limp, Trendelenburg test, bone union, length of the resected femur, limb lengthening, level of the osteotomy site, and length bridging the osteotomy site, as well as complications, were analyzed. A paired Student t-test was used to analyze continuous variables, categorical data were compared using Fisher's exact probability test, and correlation analysis was performed using Spearman's rank correlation coefficient. RESULTS: Among 14 included patients (10 females and four males), the mean age was 60.4 years (range, 47-73). The mean follow-up period was 49.1 months, and no patient was lost to follow-up. The mean Harris hip score improved from 40.7 to 87.7. The mean LLD decreased from 52 to 12.7 mm. The mean length of the excised femoral segment was 38.4 mm, and the mean length of limb lengthening was 27.1 mm. The mean distance between the osteotomy site and the lesser trochanter was 21.1 mm after surgery. The mean length of the femoral stem bridging the osteotomy site was 97.6 mm. Finally, the mean osteotomy union time was 10.6 months. No statistically significant correlation was found between the osteotomy union time and these factors. No neurological deficits were noted. Delayed union was observed in one patient, and postoperative dislocation was observed in two patients. Cement leakage into the osteotomy gap was observed in one patient, however, no revisions were required, and no signs of loosening or migration were observed. CONCLUSIONS: Cemented stem THA combined with subtrochanteric femoral shortening and transverse derotational osteotomy is safe and effective in treating patients with Crowe IV DDH. Rather than leading to nonunion, cement leakage may negatively affect bone healing.

From a Metal-Organic Square to a Robust and Regenerable Supramolecular Self-assembly for Methane Purification.

Porous supramolecular assemblies constructed by noncovalent interactions are promising for adsorptive purification of methane because of their easy regeneration. However, the poor stability arising from the weak noncovalent interactions has obstructed their practical applications. Here, we report a robust and easily regenerated polyhedron-based cationic framework assembled from a metal-organic square. This material exhibits a very low affinity for CH4 and N2 , but captures other competing gases (e.g. C2 H6 , C3 H8 , and CO2 ) with a moderate affinity. These results underpin highly selective separation of a range of gas mixtures that are relevant to natural gas and industrial off-gas. Dynamic breakthrough studies demonstrate its practical separation for C2 H6 /CH4 , C3 H8 /CH4 , CO2 /N2 , and CO2 /CH4 . Particularly, the separation time is ≈11 min g-1 for the C2 H6 /CH4 (15/85 v/v) mixture and ≈49 min g-1 for the C3 H8 /CH4 (15/85 v/v) mixture (under a flow of 2.0 mL min-1 ), respectively, enabling its capability for CH4 purification from light alkanes.

Pseudotumor and delayed recurrent dislocation after total hip arthroplasty with a modular femoral neck: A case report.

RATIONALE: Pseudotumor formation after hip arthroplasty is a rare complication that can occur not only at the head-neck junction but also at the modular neck-stem junction. Dislocation is a challenging and common complication of primary and revision total hip arthroplasty compared with other complications. Similarly, the association between pseudotumors and delayed recurrent dislocation remains unclear. PATIENT CONCERNS: We report the case of a 73-year-old woman with pseudotumor formation after total hip arthroplasty combined with a modular femoral neck. A delayed recurrent dislocation occurred in this case. Approximately 4weeks after the first revision surgery, redislocation occurred. DIAGNOSIS: The patient was eventually diagnosed with delayed recurrent artificial hip dislocation combined with a periprosthetic pseudotumor of the right hip. INTERVENTIONS: During the first revision surgery, a thickened, indurated cyst measuring 8×3×8cm with a red-brown wall containing brown fluid was completely excised. A cemented stem, combined with a BIOLOX Forte ceramic head, was implanted. Approximately 4weeks after surgery, redislocation occurred, and we cemented an elevated rim liner on the acetabular component with a metal head. OUTCOMES: At the last follow-up, 49 months after revision surgery, the patient was asymptomatic with a Harris hip score of 90. The patient had a satisfactory prognosis after treatment. LESSONS: The application of the modular-neck stem should be cautiously performed, particularly for modular prostheses containing different alloys. Pseudotumors and insufficient soft-tissue tension both contribute to hip instability, which may eventually lead to delayed repeated dislocation. In addition, femoral offset must be considered. Cement-liner technology may be used for aging patients who are less active. This case report, focusing on pseudotumors and delayed recurrent dislocations, aimed to identify factors that may support this diagnosis, which is easy to miss. Consequently, it can provide further details on the treatment process and alert orthopedic surgeons to this infrequent but important cause of delayed recurrent dislocation.

A Deep Mesencephalic Nucleus Circuit Regulates Licking Behavior / 神经科学通报·英文版

Licking behavior is important for water intake. The deep mesencephalic nucleus (DpMe) has been implicated in instinctive behaviors. However, whether the DpMe is involved in licking behavior and the precise neural circuit behind this behavior remains unknown. Here, we found that the activity of the DpMe decreased during water intake. Inhibition of vesicular glutamate transporter 2-positive (VGLUT2+) neurons in the DpMe resulted in increased water intake. Somatostatin-expressing (SST+), but not protein kinase C-δ-expressing (PKC-δ+), GABAergic neurons in the central amygdala (CeA) preferentially innervated DpMe VGLUT2+ neurons. The SST+ neurons in the CeA projecting to the DpMe were activated at the onset of licking behavior. Activation of these CeA SST+ GABAergic neurons, but not PKC-δ+ GABAergic neurons, projecting to the DpMe was sufficient to induce licking behavior and promote water intake. These findings redefine the roles of the DpMe and reveal a novel CeASST-DpMeVGLUT2 circuit that regulates licking behavior and promotes water intake.

Predictive value of systemic immune-inflammation index and sdLDL-C for contrast-induced acute kidney injury in patients with ST-segment elevation myocardial infarction undergoing emergency percutaneous coronary intervention / 中华检验医学杂志

Objective:To explore the predictive value of systemic immune-inflammation index (SII) and small and dense low-density lipoprotein-cholesterol (sdLDL-C) on contrast-induced acute kidney injury (CI-AKI) in patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary intervention (PCI).Methods:This retrospective analysis included 674 STEMI patients who underwent emergency PCI in Affiliated Hospital of Xuzhou Medical University from November 2019 to October 2021, all patients were divided into a training cohort ( n=450) and validation cohort ( n=224) at a ratio of 2∶1 according to the chronological sequence. The patients in the training cohort were further divided into CI-AKI group ( n=92) and non-CI-AKI group ( n=358). Information at admission and emergency blood biochemical indexes were collected, and the SII was calculated. Multifactorial logistic regression analysis was used to explore the independent factors influencing the occurrence of CI-AKI in STEMI patients undergoing emergency PCI in the training cohort and a predictive model was established. Receiver operating characteristic (ROC) curve and Hosmer-Lemeshow test were used to evaluate the model discrimination and calibration. Results:The prevalence of CI-AKI was 20.4% (92/450). Age, proportion of women, sdLDL-C, urea, baseline creatinine, uric acid, neutrophil count, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and SII were significantly higher in the CI-AKI group than in the non-CI-AKI group (all P<0.05), and left ventricular ejection fraction (LVEF), high-density lipoprotein cholesterol, estimated glomerular filtration rate (eGFR) and lymphocyte count were significantly lower in the CI-AKI group than in the non-CI-AKI group (all P<0.05). The results of multifactorial logistic regression analysis showed that age ( OR=1.046, P=0.001), LVEF ( OR=0.916, P<0.001), sdLDL-C ( OR=4.754, P<0.001), uric acid ( OR=1.012, P=0.007), eGFR ( OR=0.994, P=0.002), and lnSII ( OR=2.471, P<0.001) were independent determinants of CI-AKI after emergency PCI in STEMI patients. ROC curve analysis showed that area under the curve (AUC) for the diagnosis of CI-AKI was 0.688 with a sensitivity of 73.9% and specificity of 61.5% for the SII cut-off point of 1 179.07×10 9/L. The AUC for the diagnosis of CI-AKI was 0.709 with a sensitivity of 65.2% and specificity of 77.4% for the sdLDL-C cut-off point of 1.147 mmol/L. The AUC for the diagnosis of CI-AKI was 0.847 with a sensitivity of 88.0% and a specificity of 70.6% for the combination of SII and sdLDL-C with age, LVEF, uric acid and eGFR. The Hosmer-Lemeshow test (χ2=6.913, P=0.546) proved the goodness of fit of the model. Conclusions:SII and sdLDL-C have significant clinical value in the prediction of CI-AKI. SII and sdLDL-C combined with age, LVEF, uric acid and eGFR could further improve the predictive efficacy of CI-AKI.

Astaxanthin attenuates contrast-induced acute kidney injury in rats by inhibiting dynamin-related protein 1-mediated mitochondrial fission through activating SIRT1 pathway / 中华肾脏病杂志

Objective:To investigate whether astaxanthin (AST) down-regulates dynamin-related protein 1 (Drp1) through activating the silent mating type information regulation 2 homolog-1 (SIRT1) signaling pathway, thereby attenuating contrast-induced acute kidney injury.Methods:Forty adult male Sprague-Dawley rats weighing 160-180 g were randomly divided into five groups: sham surgery group (Sham group), contrast medium injury group (CM group), astaxanthin-intervention group (AST+CM group), SIRT1 inhibitor Ex527 intervention group (Ex527+CM group), and astaxanthin combined with Ex527 intervention group (AST+Ex527+CM group). After 72 hours of modeling, heart blood was removed and kidney tissues were collected for follow-up testing. Serum creatinine (Scr), blood urea nitrogen (BUN), and oxidative stress-related indexes total superoxide dismutase (T-SOD) and malondialdehyde (MDA) were measured by biochemistry; hematoxylin and eosin staining was performed to observe the pathological changes in the kidney; mitochondrial morphology and number were observed by transmission electron microscopy; reactive oxygen species (ROS) levels were detected by ROS staining in frozen sections; TUNEL staining was performed to detect apoptosis level. The expression levels of SIRT1, p53, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), Drp1 and apoptosis-related proteins Bcl-2 and Bax were detected by Western blotting.Results:(1) Compared with the CM group, Scr and BUN level were significantly lower, T-SOD level was higher and MDA level was lower in the AST+CM group, while T-SOD level decreased and MDA level increased after the combination of Ex527 (all P<0.05). (2) ROS expression was lower in the AST+CM group compared to the CM group and higher after the combination of Ex527 (both P<0.05). (3) The number of apoptotic cells was significantly reduced in the AST+CM group compared to the CM group and increased after the combination of Ex527 (both P<0.05). (4) The protein expression levels of SIRT1, PGC-1α and Bcl-2 were increased and the protein expression levels of p53, Drp1 and Bax were decreased (all P<0.05) in the AST+CM group compared with the CM group, and the protein expression levels of SIRT1, PGC-1α and Bcl-2 were decreased and the protein expression levels of p53, Drp1 and Bax were increased when Ex527 was combined (all P<0.05). Conclusion:Astaxanthin can inhibit Drp1-mediated mitochondrial fission by activating the SIRT1 pathway, thereby reducing contrast-induced acute kidney injury in rats.

[Determination of 40 dyes in oxidative hair dye products by high performance liquid chromatography].

Oxidative hair dye products have always been one of the key concerns in cosmetic supervision. The various aromatic amine and phenol dyes used in such products have different degrees of sensitization and toxicities. Therefore, it is necessary to establish a rapid and accurate method for the determination of various dyes. A high performance liquid chromatography (HPLC) method was developed for the determination of 40 dyes in oxidative hair dye products, and the method was optimized in terms of mobile phase type, column temperature, detection wavelength, and extraction solvent concentration. The sample pretreatment method was as follows: weighed 0.5 g sample accurately, added to 10 mL of sodium bisulfite solution containing 70% ethanol, extracted by ultrasonication for 15 min, added to 25 mL of the sodium bisulfite solution. Chromatographic separation was carried out on a Waters Atlantis® T3 MV Kit column (250 mm×4.6 mm, 5 µm) by gradient elution. To protect the column, the sample solution was filtered using a syringe filter with a 0.45-µm membrane before injection. The mobile phase was 0.02 mol/L ammonium acetate aqueous solution (containing 4% acetonitrile) and acetonitrile. The column temperature was varied between 30 ℃ and 35 ℃ during the separation. The dyes were detected using a photodiode array detector. The detection wavelength of 31 dyes was 235 nm, and that of the other nine dyes was 280 nm. The external standard method was used for quantitative analysis. The results showed that the 40 dyes could be separated well. All the dyes had good linearities in their own concentration ranges, with correlation coefficients (r) exceeding 0.999. The limits of detection (LODs) of the 40 dyes ranged from 5 to 168 µg/g, and the limits of quantification (LOQs) ranged from 16 to 504 µg/g. The average spiked recoveries at three levels (2.5, 5.0, and 10 mg/g) of the 40 dyes ranged from 81.4% to 109.6%, with relative standard deviations (RSDs) less than 5%. The stability of each dye in the standard solutions was good within 24 h, with RSDs ranging from 0.2% to 2.2%. To demonstrate the applicability of the method, 12 batches of commercial oxidative hair dye products from different manufacturers were analyzed. In total, 24 kinds of dyes were detected in the 12 batches of samples, and the contents of these dyes ranged from 0.01% to 2.83%. The method greatly increased the types of dyes detected by a single chromatography step, especially the permitted dyes (36 types). Compared with the standard method of Safety and Technical Standards for Cosmetics (2015 edition), this method replenished 13 permitted dyes: 1-hydroxyethyl 4,5-diaminopyrazole sulfate, hydroxyethyl-p-phenylenediaminesulfate, tetraaminopyrimidine sulfate, 2,6-dihydroxyethylaminotoluene, 2-amino-6-chloro-4-nitrophenol, 2-methyl-5-hydroxyethylaminophenol, 3-nitro-p-hydroxyethylaminophenol, 4-hydroxypropylamino-3-nitrophenol, 5-amino-4-chloro-o-cresol, 5-amino-6-chloro-o-cresol, HC yellow No. 2, hydroxybenzomorpholine, and hydroxyethyl-2-nitro-p-toluidine. The sensitivity and accuracy of the results could ensure and the detection efficiency could be improved through validation of the method. The developed method is suitable for the determination of various dyes in oxidative hair dye products and can provide an effective technical means for the supervision of such products.

miR­497/MIR497HG inhibits glioma cell proliferation by targeting CCNE1 and the miR­588/TUSC1 axis.

Emerging evidence has shown that microRNA (miR)­497 serves pivotal roles in tumorigenesis, cancer progression, metastasis and chemotherapy resistance in several types of cancer. In the present study, the expression and biological functions of miR­497 host gene (MIR497HG) were investigated in glioma tissue. The expression levels of miR­497 and MIR497HG were measured in glioma, adjacent non­cancerous and normal brain tissue and their association with the prognosis of patients with glioma were analyzed. The biological roles of miR­497 and MIR497HG were investigated in glioma cell lines. In addition, bioinformatics analysis, luciferase reporter assay and functional experiments were performed to identify and validate the downstream targets of miR­497 or MIR497HG. The expression levels of miR­497 and MIR497HG were downregulated in glioma tissue and cell lines compared with those in adjacent non­cancerous and normal brain tissue and normal human cortical neuron cell line. Patients with low miR­497 or MIR497HG expression levels exhibited a poor prognostic outcome. In addition, forced overexpression of miR­497 or MIR497HG significantly inhibited the proliferation and cell cycle progression of glioma cell lines. Furthermore, the results indicated that miR­497 and MIR497HG exerted their biological functions by direct targeting of cyclin E1 and miR­588/tumor suppressor candidate 1. In summary, the data indicated that miR­497 and MIR497HG served as tumor suppressors and may be used as potential therapeutic targets and prognostic biomarkers in glioma.

Mouse strain-specific responses of mitochondrial respiratory function and cardiac hypertrophy to isoproterenol treatment / 生理学报

Cardiac hypertrophy is a common pathological process of various cardiovascular diseases and eventually develops into heart failure. This paper was aimed to study the different pathological characteristics exhibited by different mouse strains after hypertrophy stimulation. Two mouse strains, A/J and FVB/nJ, were treated with isoproterenol (ISO) by osmotic pump to induce cardiac hypertrophy. Echocardiography was performed to monitor heart morphology and function. Mitochondria were isolated from hearts in each group, and oxidative phosphorylation function was assayed in vitro. The results showed that both strains showed a compensatory enhancement of heart contractile function after 1-week ISO treatment. The A/J mice, but not the FVB/nJ mice, developed significant cardiac hypertrophy after 3-week ISO treatment as evidenced by increases in left ventricular posterior wall thickness, heart weight/body weight ratio, cross sectional area of cardiomyocytes and cardiac hypertrophic markers. Interestingly, the heart from A/J mice contained higher mitochondrial DNA copy number compared with that from FVB/nJ mice. Functionally, the mitochondria from A/J mice displayed faster O

Molecular authentication of the medicinal species of Rhizoma et Radix Heraclei, Radix Angelicae Sinensis, Radix Angelicae Pubescentis and Rhizoma et Radix Notopterygii by integrating ITS2 and its secondary structure / 药学学报

This study is to identify Chinese medicinal materials Rhizoma et Radix Heraclei, Angelicae Sinensis, Radix Angelicae Pubescentis and Rhizoma et Radix Notopterygii based on ITS2 and its secondary structure. Total 26 ITS sequences of 7 species were downloaded from GenBank, the ITS2 sequences were annotated by HMMer method. The NJ phylogenetic tree was built by MEGA software, the intraspecific and interspecific K2P genetic distance were analyzed by MEGA as well. The ITS2 secondary structures of all taxa were predicted by ITS2 database. Sequence matrix of primary structure and secondary structure was aligned by 4Sale software. And the profile neighbor joining (PNJ) phylogenetic tree was constructed via the the ProfDistS software based on the distance method. The results show that, the average interspecific genetic distance was far greater than the average intraspecific genetic distance, an obvious barcoding gap was noted among all taxa; NJ tree showed that all species were clustered into seperate branches; each species had different secondary structures; the PNJ tree showed higher resolution than NJ tree. Therefore, ITS2 is suggested to be used as a barcode for distinguishing the original plants of Rhizoma et Radix Heraclei, Radix Angelicae Sinensis, Radix Angelicae Pubescentis and Rhizoma et Radix Notopterygii in this study, this provides some scientific basis for classification and accurate identification of these Chinese medicinal materials.

MicroRNA-144 represses gliomas progression and elevates susceptibility to Temozolomide by targeting CAV2 and FGF7.

Malignant gliomas are the most common tumor in central nervous system with poor prognosis. Due to the limitation of histological classification in earlier diagnosis and individualized medicine, it is necessary to combine the molecular signatures and the pathological characteristics of gliomas. Lots of microRNAs presented abnormal expression in gliomas and modulated gliomas development. Exploration the miRNAs profile is helpful for the diagnosis, therapy and prognosis of gliomas. It has been demonstrated that miR-144 plays important roles in solid tumors. However, the detail mechanisms remained unrevealed. In this study, we have demonstrated the level of miR-144 decreased in glioma tissues from patients, especially in gliomas with higher grades. MiR-144 was also validated have lower expression in glioma cell lines compared with cortical neuron cell by using qRT-PCR. The in vitro functional experiment indicated miR-144 improved gliomas progression through repressing proliferation, sensitizing to chemotherapeutics and inhibiting metastasis. We further identified fibroblast growth factor 7 (FGF7) and Caveolin 2 (CAV2) were target genes of miR-144 by luciferase reporter assay and western blotting. The mechanisms study suggested forced FGF7 expression elevated Akt activation and decreased reactive oxygen species (ROS) generation. The MTT and cell cycle assay indicated miR-144 suppressed glioma cells proliferation through modulating FGF mediated Akt signaling pathway. Meanwhile, miR-144 promoted Temozolomide (TMZ) induced apoptosis in glioma cells via increasing ROS production by using FACS. On the other hand, CAV2, as another target of miR-144, accelerated glioma cells migration and invasion via promoting glioma cells EMT progress. Retrieved expression of FGF7 or CAV2 rescued the proliferation and migration function mediated by miR-144. Furthermore, the in vivo experiments in PDX models displayed the anti-tumor function of miR-144, which could be retrieved by overexpression of FGF7 and CAV2. Taken together, these findings indicated miR-144 acted as a potential target against gliomas progression and uncovered a novel regulatory mechanism, which may provide a new therapeutic strategy and prognostic indicator for gliomas.

Oxidative Stress Contributes to Hyperalgesia in Osteoporotic Mice.

PURPOSE: Chronic pain is one of the most common complications of postmenopausal osteoporosis. Since oxidative stress is involved in the pathogenesis of postmenopausal osteoporosis, we explored whether oxidative stress contributes to postmenopausal osteoporotic pain. METHODS: Osteoporosis was induced in mice by ovariectomy (OVX). Pain-related behaviours were assessed by measuring sensitivity to mechanical, thermal and cold stimulation. The expression of pain-related transcripts, such acid-sensing ion channel 3 (ASIC3), transient receptor potential vanilloid 1 (TRPV1) and calcitonin gene-related peptide (CGRP), was evaluated. Plasma markers of oxidative stress were also measured. In addition, the effects of the reactive oxygen species scavenger phenyl N-tert-butylnitrone (PBN) on these parameters were assessed. RESULTS: The OVX mice presented hyperalgesia, as demonstrated by decreased paw withdrawal thresholds to mechanical stimulation and withdrawal latencies to thermal and cold stimulation, along with upregulated expression of ASIC3, TRPV1 and CGRP in the dorsal root ganglia, spinal cord and thalamus tissue. OVX elevated the plasma levels of malondialdehyde (MDA) and advanced oxidation protein products (AOPPs). However, the administration of PBN alleviated these effects. CONCLUSION: Our results indicated that oxidative stress contributes to hyperalgesia in OVX mice. Enhanced oxidative stress may be associated with osteoporotic pain. Antioxidant treatment could help alleviate chronic pain in postmenopausal osteoporotic patients.