CAT-1 as a novel CAM stabilizes endothelial integrity and mediates the protective actions of L-Arg via a NO-independent mechanism.

Interendothelial junctions play an important role in the maintenance of endothelial integrity and the regulation of vascular functions. We report here that cationic amino acid transporter-1 (CAT-1) is a novel interendothelial cell adhesion molecule (CAM). We identified that CAT-1 protein localized at cell-cell adhesive junctions, similar to the classic CAM of VE-cadherin, and knockdown of CAT-1 with siRNA led to an increase in endothelial permeability. In addition, CAT-1 formed a cis-homo-dimer and showed Ca(2+)-dependent trans-homo-interaction to cause homophilic cell-cell adhesion. Co-immunoprecipitation assays showed that CAT-1 can associate with ß-catenin. Furthermore, we found that the sub-cellular localization and function of CAT-1 are associated with cell confluency, in sub-confluent ECs CAT-1 proteins distribute on the entire surface and function as L-Arg transporters, but most of the CAT-1 in the confluent ECs are localized at interendothelial junctions and serve as CAMs. Further functional characterization has disclosed that extracellular L-Arg exposure stabilizes endothelial integrity via abating the cell junction disassembly of CAT-1 and blocking the cellular membrane CAT-1 internalization, which provides the new mechanisms for L-Arg paradox and trans-stimulation of cationic amino acid transport system (CAAT). These results suggest that CAT-1 is a novel CAM that directly regulates endothelial integrity and mediates the protective actions of L-Arg to endothelium via a NO-independent mechanism.

20(S)-Ginsenoside Rh1 alleviates sevoflurane-induced ototoxicity by reducing oxidative stress levels.

CONTEXT: Sevoflurane is an inhalational anesthetic widely used in pediatric surgery. However, animal studies have shown that multiple sevoflurane exposures during the neonatal period led to ototoxicity. 20(S)-Ginsenoside Rh1, a ginsenoside extract, protects against cisplatin-induced ototoxicity by scavenging free radicals. OBJECTIVE: This study aimed to assess the effects of Rh1 on sevoflurane-induced ototoxicity. MATERIALS AND METHODS: Neonatal cochlear explants and House Ear Institute-Organ of Corti 1 (HEI-OC1) cells were cultured and randomly divided into three groups: the control group, the sevoflurane group and the Rh1 pretreatment group. We pretreated cochlear explants or HEI-OC1 cells with 100 µM Rh1 2 hours before performing sevoflurane exposure. Immunofluorescence was used to detect hair cells and spiral ganglion neurons. Cell Counting Kit-8 assay was used to determine cell viability. Annexin V-fluorescein isothiocyanate and propidium iodide were used to evaluate apoptosis. CellROX-Green and MitoSOX-Red probes were used to measure the amount of reactive oxygen species (ROS). Tetramethylrhodamine methyl ester labeling was used to examine mitochondrial membrane potential. RESULTS: Rh1 attenuated spiral ganglion neuron nerve fibers and synapses degeneration in cochlear explants after sevoflurane exposure. Rh1 significantly increased the viability of HEI-OC1 cells, reduced reactive oxygen species accumulation in HEI-OC1 cells, and prevented mitochondrial damage in HEI-OC1 cells after sevoflurane exposure. DISCUSSION AND CONCLUSION: These findings suggest that Rh1 is a promising drug for preventing sevoflurane-induced ototoxicity.

Complete mitochondrial genomes of Dactylogyrus crucifer and Dactylogyrus zandti reveal distinct patterns of codon usage within Dactylogyrus.

Monogeneans of the genus Dactylogyrus Diesing, 1850, the largest genus in the family Dactylogyridae, mostly parasitize the gills of cyprinoid hosts; however, only 3 Dactylogyrus' mitochondrial genomes (mitogenomes) are studied so far. The aim of this research is to extend our understanding of the mitogenomes of Dactylogyrus. We sequenced the mitogenomes of D. crucifer and D. zandti isolated from Rutilus rutilus and Abramis brama orientalis in northwest China, and then we compared these mitogenomes with other monogeneans. We used Illumina NovaSeq to sequence the entire mitochondrial genomes of D. crucifer and D. zandti and characterized the mitogenomes to understand the gene structure, gene identity, the secondary structures of the 22 tRNA genes, and relative synonymous codon usage. We used the analytic Bayesian Information and Maximum Likelihood methods to determine their associated phylogenetic trees. The mitogenomes of D. crucifer and D. zandti were 14,403 and 18,584 bp, respectively. Organization and positioning of these genes were in accordance with Dactylogyrus lamellatus and Dactylogyrus tuba. The nucleotide composition of Dactylogyridae was different from other families of Monogenea, and the A+T count of genus Dactylogyrus (54 - 58.4 %) was lower than other genus species of the family Dactylogyridea (63.9 - 78.4 %) in protein-coding genes. Dactylogyrus members displayed a codon usage bias. The relative synonymous codon used by Dactylogyrus was not conserved and was lower than other monogeneans. The codon use patterns of closely-related species isolated from closely-related hosts were identical. Phylogenetic analyses using mitogenomic dataset produced Dactylogyrus isolated from host subfamily Leuciscinae formed a sister-group. Our results contributed significantly to an increased database of mitogenomes, more than 50 %, for Dactylogyrus that may help future studies of mitochondrial genes and codon uses for the analysis of monogenean phylogenetics.

Enhanced phosphorylation of caveolar PKC-α limits peptide internalization in lung endothelial cells.

We previously reported that the vasoactive peptide 1 (P1, "SSWRRKRKESS") modulates the tension of pulmonary artery vessels through caveolar endothelial nitric oxide synthase (eNOS) activation in intact lung endothelial cells (ECs). Since PKC-α is a caveolae resident protein and caveolae play a critical role in the peptide internalization process, we determined whether modulation of caveolae and/or caveolar PKC-α phosphorylation regulates internalization of P1 in lung ECs. Cell monolayers were incubated in culture medium containing Rhodamine red-labeled P1 (100 µM) for 0-120 min. Confocal examinations indicate that P1 internalization is time-dependent and reaches a plateau at 60 min. Caveolae disruption by methyl-ß-cyclodextrin (CD) and filipin (FIL) inhibited the internalization of P1 in ECs suggesting that P1 internalizes via caveolae. P1-stimulation also enhances phosphorylation of caveolar PKC-α and increases intracellular calcium (Ca(2+)) release in intact cells suggesting that P1 internalization is regulated by PKC-α in ECs. To confirm the roles of increased phosphorylation of PKC-α and Ca(2+) release in internalization of P1, PKC-α modulation by phorbol ester (PMA), PKC-α knockdown, and Ca(2+) scavenger BAPTA-AM model systems were used. PMA-stimulated phosphorylation of caveolar PKC-α is associated with significant reduction in P1 internalization. In contrast, PKC-α deficiency and reduced phosphorylation of PKC-α enhanced P1 internalization. P1-mediated increased phosphorylation of PKC-α appears to be associated with increased intracellular calcium (Ca(2+)) release since the Ca(2+) scavenger BAPTA-AM enhanced P1 internalization. These data indicate that caveolar integrity and P1-mediated increased phosphorylation of caveolar PKC-α play crucial roles in the regulation of P1 internalization in lung ECs.

Aldehyde dehydrogenase 2 and PARP1 interaction modulates hepatic HDL biogenesis by LXRα-mediated ABCA1 expression.

HDL cholesterol (HDL-C) predicts risk of cardiovascular disease (CVD), but the factors regulating HDL are incompletely understood. Emerging data link CVD risk to decreased HDL-C in 8% of the world population and 40% of East Asians who carry an SNP of aldehyde dehydrogenase 2 (ALDH2) rs671, responsible for alcohol flushing syndrome; however, the underlying mechanisms remain unknown. We found significantly decreased HDL-C with increased hepatosteatosis in ALDH2-KO (AKO), ALDH2/LDLR-double KO (ALKO), and ALDH2 rs671-knock-in (KI) mice after consumption of a Western diet. Metabolomics identified ADP-ribose as the most significantly increased metabolites in the ALKO mouse liver. Moreover, ALDH2 interacted with poly(ADP-ribose) polymerase 1 (PARP1) and attenuated PARP1 nuclear translocation to downregulate poly(ADP-ribosyl)ation of liver X receptor α (LXRα), leading to an upregulation of ATP-binding cassette transporter A1 (ABCA1) and HDL biogenesis. Conversely, AKO or ALKO mice exhibited lower HDL-C with ABCA1 downregulation due to increased nuclear PARP1 and upregulation of LXRα poly(ADP-ribosyl)ation. Consistently, PARP1 inhibition rescued ALDH2 deficiency-induced fatty liver and elevated HDL-C in AKO mice. Interestingly, KI mouse or human liver tissues showed ABCA1 downregulation with increased nuclear PARP1 and LXRα poly(ADP-ribosyl)ation. Our study uncovered a key role of ALDH2 in HDL biogenesis through the LXRα/PARP1/ABCA1 axis, highlighting a potential therapeutic strategy in CVD.

Pharmacological profiles of the murine gastric and colonic H,K-ATPases.

BACKGROUND: The H,K-ATPase, consisting of α and ß subunits, belongs to the P-type ATPase family. There are two isoforms of the α subunit, HKα1 and HKα2 encoded by different genes. The ouabain-resistant gastric HKα1-H,K-ATPase is Sch28080-sensitive. However, the colonic HKα2-H,K-ATPase from different species shows poor primary structure conservation of the HKα2 subunit between species and diverse pharmacological sensitivity to ouabain and Sch28080. This study sought to determine the contribution of each gene to functional activity and its pharmacological profile using mouse models with targeted disruption of HKα1, HKα2, or HKbeta genes. METHODS: Membrane vesicles from gastric mucosa and distal colon in wild-type (WT), HKα1, HKα2, or HKß knockout (KO) mice were extracted. K-ATPase activity and pharmacological profiles were examined. RESULTS: The colonic H,K-ATPase demonstrated slightly greater affinity for K(+) than the gastric H,K-ATPase. This K-ATPase activity was not detected in the colon of HKα2 KO but was observed in HKß KO with properties indistinguishable from WT. Neither ouabain nor Sch28080 had a significant effect on the WT colonic K-ATPase activity, but orthovanadate abolished this activity. Amiloride and its analogs benzamil and 5-N-ethyl-N-isopropylamiloride inhibited K-ATPase activity of HKα1-containing H,K-ATPase; the dose dependence of inhibition was similar for all three inhibitors. In contrast, the colonic HKα2-H,K-ATPase was not inhibited by these compounds. CONCLUSIONS: These data demonstrate that the mouse colonic H,K-ATPase exhibits a ouabain- and Sch28080-insensitive, orthovanadate-sensitive K-ATPase activity. Interestingly, pharmacological studies suggested that the mouse gastric H,K-ATPase is sensitive to amiloride. GENERAL SIGNIFICANCE: Characterization of the pharmacological profiles of the H,K-ATPases is important for understanding the relevant knockout animals and for considering the specificity of the inhibitors.

[Clinical application of nose ring drain technique combined with Ilizarov circular external fixation for Gustilo â ¢A Pilon fractures].

OBJECTIVE: To investigate the effectiveness of the nose ring drain (NRD) technique combined with Ilizarov circular external fixation in treatment of Gustilo ⅢA Pilon fracture. METHODS: Between March 2017 and December 2019, 17 patients with Gustilo ⅢA Pilon fractures were admitted and treated with NRD technique combined with Ilizarov circular external fixation. Among them, there were 11 males and 6 females; the age ranged from 24 to 63 years, with an average of 38.2 years. There were 3 cases of traffic accident injury, 13 cases of falling injury, and 1 case of penetrating injury. There were 13 cases of emergency admittance and 4 cases of wound infection after surgical treatment. Furthermore, there were 2 cases of fibula fractures and 3 cases of lateral malleolus fractures. RESULTS: All patients were followed up 8-12 months, with an average of 9.9 months. All wounds healed by first intention, and 4 patients with preoperative infection had no recurrence during the follow-up. The external fixator was removed after fracture healing in 17 patients at 3-7 months after operation (mean, 4.5 months). At last follow-up, the pain score of the ankle joint Kofoe score was 40-50, with an average of 44; the functional score was 17-27, with an average of 25; the mobility score was 8-18, with an average of 14; and the effectiveness was rated as excellent in 8 cases, good in 7 cases, and poor in 1 case. CONCLUSION: For Gustilo ⅢA Pilon fractures, the NRD technique combined with Ilizarov circular external fixation has advantages of good fracture fixation and drainage effects, which greatly reduces the complications of traditional treatment options and the number of operations.

Heterogeneity of H-K-ATPase-mediated acid secretion along the mouse collecting duct.

In the collecting duct (CD), H-K-ATPases function in cation reabsorption and H secretion. This study evaluated H-K-ATPase-mediated H secretion along the mouse CD, measured as EIPA- and luminal bafilomycin A(1)-insensitive intracellular pH (pH(i)) recovery from acute H loading (NH(4)) using BCECF. pH(i) recovery was measured in 1) microperfused cortical, outer medullary, and inner medullary CDs (CCD, OMCD, and IMCD) from C57BL/6J mice fed a normal diet and 2) common murine CD cell lines. H-K-ATPase activity along the native, microperfused CD was greatest in the CCD, less in the OMCD, and least in the IMCD (0.10 +/- 0.02, 0.04 +/- 0.01, and 0.01 +/- 0.002 U/min, respectively). H-K-ATPase activity was 0.30 +/- 0.03 and 0.26 +/- 0.03 in A- and B-type ICs, respectively, and was sensitive to Sch-28080 or ouabain. pH(i) recovery was greatest in the OMCD(1) cell line (0.25 +/- 0.01) and less in mpkCCD(c14) (0.17 +/- 0.01), mIMCD-K2 (0.12 +/- 0.01), and mIMCD-3 (0.05 +/- 0.01) cells. EIPA inhibited the majority of pH(i) recovery in these cells (100%, 64%, 75%, and 80% in mpkCCD(c14), OMCD(1), mIMCD-K2, and mIMCD-3, respectively). In OMCD(1) cells, where EIPA-insensitive pH(i) recovery was greatest, H-K-ATPase activity was 0.10 +/- 0.01 and was significantly inhibited (80%) by Sch-28080. We conclude that 1) H-K-ATPase-mediated H secretion in the native mouse CD is greatest in the ICs of the CCD, 2) A- and B-type ICs possess HKalpha(1) and HKalpha(2) H-K-ATPase activity, and 3) the OMCD(1) cell line best exhibits H-K-ATPase.

[Application of simple Ilizarov ring external fixation technique in treatment of tibial plateau fracture complicated with osteofascial compartment syndrome].

OBJECTIVE: To explore the effectiveness of simple Ilizarov ring external fixation technique in treatment of tibial plateau fractures complicated with osteofascial compartment syndrome. METHODS: Between September 2013 and March 2017, 30 patients with tibial plateau fractures complicated with osteofascial compartment syndrome were treated with simple Ilizarov ring external fixation technique. There were 23 males and 7 females, with an average age of 34.4 years (range, 23-43 years). The injuries were caused by traffic accident in 12 cases, by falling from height in 4 cases, by falling in 8 cases, and by a crashing object in 6 cases. The time from injury to admission was 1-12 hours (mean, 4.8 hours). According to the Schatzker classification, there was 1 case of type Ⅱ, 3 cases of type Ⅲ, 10 cases of type Ⅳ, 7 cases of type Ⅴ, and 9 cases of type Ⅵ. All patients underwent fasciotomy due to osteofascial compartment syndrome; the interval between fasciotomy and operation was 10-15 days (mean, 12.5 days). Knee Society Score (KSS) and Ilizarov Method Research and Application Association (ASAMI) protocol were used to evaluate knee function. RESULTS: The operation time was 110-155 minutes (mean, 123.1 minutes); the intraoperative blood loss was 100-500 mL (mean, 245 mL); the postoperative hospital stay was 3-5 days (mean, 3.8 days). All patients were followed up 20-24 weeks (mean, 22.7 weeks). Except for 2 patients with signs of needle tract infection, no other complication occurred. X-ray films showed that the fractures healed, and the healing time was 10-20 weeks (mean, 14.6 weeks). At last follow-up, the KSS clinical score was 70- 95 with an average of 87.5; the functional score was 70-90 with an average of 79.0. According to ASAMI protocol evaluation, the effectiveness was rated as excellent in 24 cases, good in 3 cases, fair in 2 cases, and poor in 1 case. CONCLUSION: For tibial plateau fractures complicated with osteofascial compartment syndrome, simple Ilizarov ring external fixation technique can basically restore joint function and has fewer complications. It is a relatively safe and effective treatment method.

[Primarily application of Ilizarov microcirculation reconstruction technique for chronic wounds in post-traumatic ischemia limbs].

OBJECTIVE: To evaluate the treatment results of Ilizarov microcirculation reconstruction technique for chronic wounds in the post-traumatic ischemia limbs. METHODS: Between January 2016 and July 2019, 7 cases of chronic wounds in the post-traumatic ischemia limbs were treated. There were 5 males and 2 females, with an average age of 42.4 years (range, 29-66 years). The duration of the wound ranged from 1 month to 2 years (mean, 7.7 months). The wounds located in the leg (3 cases) or in the foot and ankle (4 cases). The wound sizes ranged from 4.0 cm×2.2 cm to 12.0 cm×7.1 cm. There were 1 case of tibial varus, 3 cases of equinovarus, 1 case of scleroderma, and 2 cases of Volkmann's ischemic contracture. After debridement, external fixators were used for tibial transverse transport, or correction of tibial varus and correction of equinovarus. RESULTS: All patients were followed up 8-20 months, with an average of 13 months. The infection of wound surface was all controlled in 7 cases and the granulation tissue grew well; the wound surface healed directly in 5 cases and healed after skin grafting in 2 cases, and the wound healing time was 1-3 months (mean, 1.7 months). During the follow-up, there was no recurrence of the wound. Six cases of limb deformity were corrected. CONCLUSION: For the chronic wounds in the post-traumatic ischemia limbs, Ilizarov microcirculation reconstruction technique can effectively improve local circulation and facilitate the fresh granule growth and wound healing.

[Anatomical calcaneal external fixator self-designed according to the morphology of heel].

OBJECTIVE: The anatomical calcaneal external fixator was designed by measuring and calculating the morphological data of the heel. METHODS: A total of 100 normal people were randomly selected to obtain 200 hind foot data, including 45 males and 55 females, with an average age of 43.9 years (range, 19-67 years). According to the principles of human engineering and local anatomy, the morphological data of the heel in the weight-bearing standing position and supine position were measured with the direct measurement mode. The heel length, heel width, heel height, medial ankle height, lateral ankle height, and calcaneal pitch angle (CPA) were measured by vernier calipers and ulnar markers in weight-bearing standing position, and the gender groups and left and right foot groups were compared; the shape of the hind foot in the supine position was measured by three-dimensional (3D) dot matrix inverse model method. According to the stereoscopic data of the comprehensive anatomical morphology of the heel, the anatomical calcaneal external fixator was designed with AutoCAD 2019 and other 3D industrial design softwares. RESULTS: The measurements of shoe size, heel length, heel width, heel height, medial ankle height, lateral ankle height, and CPA in male were significantly higher than those in female ( P<0.05). There was no significant difference between the left and right feet in the other indexes except that the height of the medial malleolus of the left foot was significantly lower than that of the right foot ( t=-2.827, P=0.005). The measurement of 3D dot matrix inverse model in supine position showed that the heel part was non-circular arc edge, and many groups of arc edges fluctuate in a limited range. Based on the above data, an anatomical calcaneal external fixator was designed, which could fit the anatomic radian in theory, so as to be flexible in configuration. On this basis, the ordinary configuration, compression configuration, and orthodontic configuration were designed to meet the treatment needs of calcaneal fractures in different degrees. The ordinary configuration was suitable for patients with Sanders Ⅰ, ⅡA, and ⅡB calcaneal fractures with no or slight displacement of intra-articular fractures; the ordinary configuration was mainly used for simple fixing. The compression configuration was suitable for patients with Sanders ⅡC, ⅢA, and ⅢB, tongue fractures, and avulsion fractures with severe displacement of intra-articular fractures; the compression configuration used obliquely drawn console wires to fix the displaced bones. The orthodontic configuration was suitable for patients with Sanders ⅢC and Ⅳ calcaneal fractures with severe displacement of intra-articular fractures or severe calcaneal bone defects; the orthodontic configuration was a multi-module design, which took into account the stable fixation of the fracture and the arbitrary adjustment of the joint fixation angle. CONCLUSION: The hind foot is special for morphology, so the external fixator designed based on the vernier caliper measurement method and 3D dot matrix measuring plate measurement method is an anatomical type and its configuration can theoretically meet stable and flexible clinical needs.

Sarcomere mechanics in capillary endothelial cells.

Tension generation in endothelial cells of the aorta, spleen, and eye occurs in actin stress fibers, and is necessary for normal cell function. Sarcomeres are the tension-generating units of actin stress fibers in endothelial cells. How sarcomeres generate and maintain tension in stress fibers is not well understood. Using femtosecond laser ablation, we severed living stress fibers and measured sarcomere contraction under zero tension. The length of the sarcomere decreased in two phases: an instantaneous initial response, followed by a slower change in length attributed to myosin activity. The latter phase ceased abruptly after a minimum sarcomere length was reached, suggesting a rigid resistance that prevents further contraction. Furthermore, severed, contracted stress fibers did not relax when treated with myosin inhibitors, indicating that contracted stress fibers do not store elastic potential energy. These novel measurements combined with modeling suggest that myosin-generated forces in adjacent sarcomeres are directly in balance, and argue against sarcomere models with springlike elements in parallel with myosin contractile elements. We propose a new model for tension generation in the sarcomere, which provides a mechanistic interpretation for our observations and previous observations of inhomogeneous sarcomere contraction and apparent stress fiber viscoelastic behavior.

Substance P increases cell-surface expression of CD74 (receptor for macrophage migration inhibitory factor): in vivo biotinylation of urothelial cell-surface proteins.

Macrophage migration inhibitory factor (MIF), an inflammatory cytokine, and its receptor CD74 are upregulated by bladder inflammation. MIF-mediated signal transduction involves binding to cell-surface CD74, this study documents, in vivo, MIF-CD74 interactions at the urothelial cell surface. N-hydroxysulfosuccinimide biotin ester-labeled surface urothelial proteins in rats treated either with saline or substance P (SP, 40 microg/kg). The bladder was examined by histology and confocal microscopy. Biotinylated proteins were purified by avidin agarose, immunoprecipitated with anti-MIF or anti-CD74 antibodies, and detected with strepavidin-HRP. Only superficial urothelial cells were biotinylated. These cells contained a biotinylated MIF/CD74 cell-surface complex that was increased in SP-treated animals. SP treatment increased MIF and CD74 mRNA in urothelial cells. Our data indicate that intraluminal MIF, released from urothelial cells as a consequence of SP treatment, interacts with urothelial cell-surface CD74. These results document that our previously described MIF-CD74 interaction occurs at the urothelial cell surface.

The effect of esketamine on postoperative recovery in children after endoscopic adenoidectomy / 复旦学报（医学版）

Objective To observe the effect of esketamine on postoperative recovery in children after endoscopic adenoidectomy.Methods Sixty pediatric patients who underwent adenoidectomy with endoscope from Jan 2022 to Jan 2023 in Eye&ENT Hospital,Fudan University were enrolled.The pediatric patients were randomly divided into hydro-morphine group(n=30)and esketamine group(n=30).Anesthesia induction:lidocaine 1.5 mg/kg,propofol 2.5 mg/kg and remifentanil 4 μg/kg were injected intravenously,and then the endotracheal tube was used for airway management.Anesthesia maintenance:remifentanil infusion was at 0.2-0.5 μg·kg-1·min-1 and the end tidal concentration of sevoflurane was at 0.7-1.0 minimum alveolar concentration(MAC).At the end of surgery,either hydromorphone 0.01 mg/kg or esketamine 0.5 mg/kg were administered for postoperative pain control.Time to resume spontaneous breathing was recorded.Other parameters included respiratory rate per minute,duration of stay in the post-anesthesia care unit,hemodynamic profiles.The adverse events including agitation and desaturation were also of note.Results Children in esketamine group resumed spontaneous breathing faster(P=0.048),had faster respiratory rate when recovery of spontaneous breathing(P=0.001)and lower concentration of end tidal CO2(P=0.005).The findings suggested that esketamine did not impair respiratory function.Compared to hydro-morphine group,children in esketamine group had shorter stay in the post-anesthesia care unit with statistical difference(P=0.020).Esketamine had no effect on heart rate and blood pressure,so there were less adverse events.Conclusion Compared with 0.01 mg/kg hydro-morphine,0.5 mg/kg esketamine does not impair respiratory function and it facilitate fast recovery in children undergoing endoscopic adenoidectomy after general anesthesia.

Quercetin plays a neuroprotective role in inhibiting mitochondrial apoptosis by mediating JNK signaling pathway / 中国药理学通报

Aim To study the mechanism of quereetin (Que) inhibiting mitochondrial damage induced by Aβ

Simultaneous content determination of twelve constituents in Bushen Huoxue Sanjie Capsules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of gallic acid,protocatechuic acid,morroniside,loganin,sweroside,paeoniflorin,hypericin,astragalin,salvianolic acid B,salvianolic acid A,epimedin C and icariin in Bushen Huoxue Sanjie Capsules.METHODS The analysis was performed on a 30℃thermostatic Agilent 5 TC-C18 column(250 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 240 nm.RESULTS Twelve constituents showed good linear relationships within their own ranges(r≥0.999 8),whose average recoveries were 97.11%-101.14%with the RSDs of 0.60%-2.65%.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Bushen Huoxue Sanjie Capsules.

Mutation characteristics of angioimmunoblastic T-cell lymphoma: an analysis of 75 cases / 中华病理学杂志

Objective: To investigate the characteristics of gene mutations in angioimmunoblastic T-cell lymphoma (AITL). Methods: Seventy-five AITL cases diagnosed at the Department of Pathology, Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China from June 2021 to June 2023 were included. Their formalin-fixed and paraffin-embedded or fresh tissues were subject to targeted next generation sequencing (NGS). The sequencing data was collected, and the distribution and type of gene mutations were analyzed. Results: 492 potential driver mutations were identified in 74 out of the 84 genes. Targeted sequencing data for the 75 AITL patients showed that the genes with mutation frequencies of ≥10% were TET2 (89.3%), RHOA (57.3%), IDH2 (37.3%), DNMT3A (36.0%), KMT2C (21.3%), PLCG1 (12.0%), and KDM6B (10.7%). There were significant co-occurrence relationships between TET2 and RHOA, TET2 and IDH2, and RHOA and IDH2 gene mutations (P<0.05), respectively, while TET2 and KDM6B gene mutations were mutually exclusive (P<0.05). Conclusions: The study reveals the mutational characteristics of AITL patients using NGS technology, which would provide insights for molecular diagnosis and targeted therapy of AITL.

Diquafosol sodium combined with intense pulsed light in the treatment of meibomian gland dysfunction / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the effect of 3% diquafosol sodium eye drops combined with intense pulsed light on the treatment of meibomian gland dysfunction and the change of meibomian glands.METHODS: Prospective study. A total of 141 patients(282 eyes)who were diagnosed with meibomian gland dysfunction from January 2021 to May 2022 in our hospital were selected and they were randomly divided into the control group(73 cases, 146 eyes)and the observation group(68 cases, 136 eyes)according to random number table. The control group was given 0.3% sodium hyaluronate eye drops combined with intense pulsed light, and the observation group was treated with 3% diquafosol sodium eye drops combined with intense pulsed light. The subjective symptom score, physical sign score, non-invasive tear break-up time, tear meniscus height, lipid layer thickness, and meibomian gland density before and after the treatment were compared between the two groups at 2wk after the end of treatment.RESULTS: There were no differences in the subjective symptom score, physical sign score, non-invasive tear break-up time, tear meniscus height, lipid layer thickness, and meibomian gland density between the two groups of patients before treatment(P&#x003E;0.05). After 2wk of treatment, the symptom scores and physical sign scores of patients in the two groups continued to decrease, non-invasive tear break-up time and lipid layer thickness continued to increase, and the meibomian gland density also increased. The tear meniscus height in the observation group increased, while the control group showed no significant changes. The observation group had better clinical indicators than the control group(P&#x003C;0.05). No obvious complications were observed in all patients.CONCLUSION: The combination of diquafosol sodium eye drops and intense pulsed light is synergistic in the treatment of meibomian gland dysfunction, with significant therapeutic effects and improvement of meibomian gland repair, which is significantly superior to simple intense pulsed light therapy.

Clinicopathological and prognostic features of subungual melanoma in situ / 中华病理学杂志

Objective: To investigate the clinicopathological characteristics, immunohistochemical profiles, molecular features, and prognosis of subungual melanoma in situ (SMIS). Methods: Thirty cases of SMIS were collected in Fudan University Shanghai Cancer Center, Shanghai, China from 2018 to 2022. The clinicopathological characteristics and follow-up data were retrospectively analyzed. Histopathologic evaluation and immunohistochemical studies were carried out. By using Vysis melanoma fluorescence in situ hybridization (FISH) probe kit, combined with 9p21(CDKN2A) and 8q24(MYC) assays were performed. Results: There were 8 males and 22 females. The patients' ages ranged from 22 to 65 years (median 48 years). All patients presented with longitudinal melanonychia involving a single digit. Thumb was the most commonly affected digit (16/30, 53.3%). 56.7% (17/30) of the cases presented with Hutchinson's sign. Microscopically, melanocytes proliferated along the dermo-epithelial junction. Hyperchromatism and nuclear pleomorphism were two of the most common histological features. The melanocyte count ranged from 30 to 185. Most cases showed small to medium nuclear enlargement (29/30, 96.7%). Pagetoid spread was seen in all cases. Intra-epithelial mitoses were identified in 56.7% (17/30) of the cases. Involvement of nailfold was found in 19 cases, 4 of which were accompanied by cutaneous adnexal extension. The positive rates of SOX10, PNL2, Melan A, HMB45, S-100, and PRAME were 100.0%, 100.0%, 96.0%, 95.0%, 76.9%, and 83.3%, respectively. FISH analysis was positive in 6/9 of the cases. Follow-up data were available in 28 patients, and all of them were alive without disease. Conclusions: SMIS mainly shows small to medium-sized cells. High melanocyte count, hyperchromatism, nuclear pleomorphism, Pagetoid spreading, intra-epithelial mitosis, nailfold involvement, and cutaneous adnexal extension are important diagnostic hallmarks. Immunohistochemistry including SOX10 and PRAME, combined with FISH analysis, is valuable for the diagnosis of SMIS.

Mechanism of Yougui Pills in the treatment of knee osteoarthritis based on network pharmacology and molecular docking / 西安交通大学学报(医学版)

【Objective】 To explore the mechanism of Yougui Pills in the treatment of knee osteoarthritis （KOA） based on network pharmacology and molecular docking. 【Methods】 The active components of Yougui Pills were obtained by searching TCMSP database. With the active component targets predicted by TCMSP， SwissTargetPrediction and TargetNet， we searched TCMIP， CTD， DisGeNET and GeneCards databases for KOA related targets. By crossing the two sets of targets， we obtained therapeutic targets of Yougui Pills for KOA. FDA-approved drugs for KOA and their targets were searched in the DrugBank database， and the drug-target network was constructed by using Cytoscape. Then the protein-protein interaction （PPI） network was constructed by using STRING database. After Go and KEGG enrichment analyses， the network of “traditional Chinese Medicine-active components-key targets-significant pathways-biological process” was constructed. 【Results】 A total of 132 active components were obtained from the nine herbal medicines of Yougui Pills， corresponding to 490 targets， of which 76 were potential therapeutic targets. Of them 13 were the same as the targets of 5 Western medicines for KOA. We screened out 18 key targets by the PPI network. Go enrichment analysis revealed 1281 biological processes， 60 molecular functions， and 17 cellular components， which were mainly related to inflammation， infection and oxidative stress. KEGG enrichment analysis showed that 50 pathways were significantly enriched， including IL-17 signaling pathway and TNF signaling pathway， whose functions were mainly involved in immune system， signal transduction， endocrine system， cell growth and death and other biological processes. 【Conclusion】 With multiple compounds acting on multiple molecular targets， Yougui Pills can treat KOA through molecular mechanisms including inhibiting inflammation， reducing oxidative stress， and promoting cartilage cell proliferation.