miR-5581 Contributes to Osteoarthritis by Targeting NRF1 to Disturb the Proliferation and Functions of Chondrocytes.

Chondrocyte survival is critical for the preservation of a healthy cartilage matrix. Limited chondrocyte function and survival can result in articular cartilage failure, thereby contributing to osteoarthritis (OA). In this study, miR-5581 was significantly up-regulated in OA samples, and miR-5581-associated genes were enriched in Kras signaling. miR-5581 up-regulation was observed in clinical OA samples and IL-1ß-stimulated chondrocytes. miR-5581 inhibition attenuated IL-1ß-induced chondrocyte proliferation suppression, extracellular matrix (ECM) synthesis suppression and degradation, and IL-1ß-suppressed Kras signaling activation. miR-5581 was targeted to inhibit NRF1. In IL-1ß-treated chondrocytes, NRF1 overexpression attenuated IL-1ß-induced cellular damage and partially abolished the effects of miR-5581 overexpression on IL-1ß-stimulated chondrocytes. NRF1 was down-regulated in knee joint cartilage of OA mice. In conclusion, miR-5581, which was up-regulated in OA samples and IL-1ß-stimulated chondrocytes, inhibited chondrocyte proliferation and ECM synthesis, and promoted ECM degradation through targeting NRF1, whereby Kras signaling might be involved.

Efficacy of a modified needle grasper for single-port laparoscopic hernia repair in children: a propensity score-matched analysis.

PURPOSE: The advantage of using the single-port technique over the conventional two-port approach is uncertain. This study aimed to evaluate the outcomes of a single-port laparoscopic percutaneous extraperitoneal closure (SLPEC) using a modified needle grasper in children and compare the results to those of two-port laparoscopic percutaneous extraperitoneal closure (TLPEC). METHODS: A retrospective cohort analysis of SLPEC and TLPEC surgery from February 2016 to June 2021 was conducted at our institution. Pediatric patients underwent SLPEC using the modified needle grasper to complete the high ligation of the hernia sac, while operations in the conventional two-port group only used regular laparoscopic instruments. A 1:1 propensity score matching (PSM) analysis was used to reduce selection bias. RESULTS: Of 1320 patients, 1169 were included in the single-port/two-port crude evaluation, with 930 in the PSM cohort (465 patients/arm). Among 1:1 matched patients, the operation time for single-port patients vs. two-port patients were 11.28 ± 3.98 vs. 15.47 ± 4.54 min for unilateral repair and 16.86 ± 4.59 vs. 20.40 ± 4.29 min for bilateral repair (p < .05). Cosmetic results did not differ between the SLPEC and TLPEC groups (0% vs. 0.7%, p = 0.249). The recurrence rates were comparable between the two groups (0.6% vs. 1.1%, p = 0.725). Moreover, the differences in surgical site infection (SSI), testicular atrophy, open conversion and postoperative hydrocele occurrence were insignificant between the two groups. CONCLUSIONS: In this cohort study, the modified needle grasper is a safe and feasible instrument for SLPEC, and SLPEC using the needle grasper has a shorter operation time than TLPEC.

Caffeine alleviates acute liver injury by inducing the expression of NEDD4L and deceasing GRP78 level via ubiquitination.

BACKGROUND: Acute liver injury is liver cell injury that occurs rapidly in a short period of time. Caffeine has been shown to maintain hepatoprotective effect with an unclear mechanism. Endoplasmic reticulum stress (ERS) has significant effects in acute liver injury. Induction of GRP78 is a hallmark of ERS. Whether or not caffeine's function is related to GRP78 remains to be explored. METHODS: Acute liver injury model was established by LPS-treated L02 cells and in vivo administration of LPS/D-Gal in mice. Caffeine was pre-treated in L02 cells or mice. Gene levels was determined by real-time PCR and western blot. Cell viability was tested by CCK-8 assay and cell apoptosis was tested by flow cytometry. The interaction of GRP78 and NEDD4L was determined by Pull-down and co-immunoprecipitation (Co-IP) assay. The ubiquitination by NEDD4L on GRP78 was validated by in vitro ubiquitination assay. RESULTS: Caffeine protected liver cells against acute injury induced cell apoptosis and ERS both in vitro and in vivo. Suppression of GRP78 could block the LPS-induced cell apoptosis and ERS. NEDD4L was found to interact with GRP78 and ubiquitinate its lysine of 324 site directly. Caffeine treatment induced the expression of NEDD4L, resulting in the ubiquitination and inhibition of GRP78. CONCLUSION: Caffeine mitigated the acute liver injury by stimulating NEDD4L expression, which inhibited GRP78 expression via ubiquitination at its K324 site. Low dose of caffeine could be a promising therapeutic treatment for acute liver injury.

Therapeutical plasma exchange for thrombotic thrombocytopenic purpura in the emergency department: A single center experience.

INTRODUCTION: Thrombotic thrombocytopenic purpura (TTP) is a rare, life-threatening and easily misdiagnosed thrombotic microangiopathy disease. Few studies have reported the use of therapeutic plasma exchange (TPE) for TTP in emergency departments in China. The present study was a retrospective analysis of patients with TTP who were treated with TPE in our emergency intensive care unit (EICU). METHODS: This study retrospectively analyzed patients with TTP who received TPE management from July 1, 2014 to February 1, 2020. The following clinical data of these patients were collected: laboratory results, first symptoms, ADAMTS13 levels, glucocorticoid levels, TPE times and outcomes. RESULTS: The study included 19 patients (9 male and 10 female) with 20 clinical episodes, and 1 female patient had two episodes. TPE was used in 17 patients, and TPE was performed once every 2-3 days in patients. The volume for each TPE treatment was 2000 ml. In total, 4 male patients died, and 15 patients survived. One female experienced a relapse. No significant differences in age, RBC, HGB, PLT, ALT, AST, BUN, Cr, LDH, or bilirubin were noted between the survival and death groups. The mortality rate of male patients was significantly higher than that of female patients(p = 0.0325, p < 0.05), and the mean age of deceased patients was 64.25 ± 4.78 years, which was older than the mean age of survivors (47.38 ± 4.30). However, no significant difference was noted (p = 0.0787). CONCLUSION: TPE had satisfactory results for TTP patients although it was not performed every day. Older male TTP patients exhibited a relatively increased risk of death.

Effect of ligand-receptor interaction of osteopontin-CD44 on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes in vitro. / 骨桥蛋白-CD44é ä½“å—ä½“ååº”å¯¹äººè†éª¨å ³èŠ‚ç‚Žè½¯éª¨ç»†èƒžé€æ˜Žè´¨é ¸è¡¨è¾¾çš„å½±å“.

OBJECTIVES: To investigate the effect of ligand-receptor interaction of osteopontin (OPN)-CD44 on the expression of hyaluronic acid (HA) in the cultured human knee osteoarthritis (OA) chondrocytes via interfering the reaction between OPN and CD44 ligand-receptor. METHODS: The OA chondrocytes and normal chondrocytes were obtained from knee joint cartilage tissues in the patients with knee OA and malignant tumor respectively. The normal chondrocytes and OA chondrocytes were detected and analyzed, and then the intervention analysis of OA chondrocytes was carried out. The OA chondrocytes were divided into 4 groups: a negative control group, which was cultured with complete medium without any molecular intervention reagent; an OPN intervention group, which was cultured with recombinant human OPN (rhOPN) for 24 hours; a CD44 blocking group, which were pretreated with CD44 receptor specific antagonist for 1 hour to block the binding of OPN-CD44, and then treated with rhOPN for 23 hours; a CD44 homotype group, which was pretreated with CD44 for 1 hour and then treated with rhOPN for 23 hours. In addition, the study for OPN-CD44 axis was also divided into 4 groups: an OA-negative control group (OA-NC group), a si-OPN intervention group, a rhOPN intervention group, and a rhOPN + CD44 antibody (Ab) group. Western blotting, real-time PCR, and enzyme linked immunosorbent assay (ELISA) were used to detect the protein and mRNA expression levels of OPN, CD44, hyaluronate synthase (HAS), and HA, respectively. RESULTS: The protein expression levels of OPN, CD44, and HAS1 and the secretion levels of HA in the OA chondrocytes were higher than those in the normal chondrocytes. Compared with the OPN intervention group, the expression levels of HAS1, HAS2, HAS3 and HA in the CD44 blocking group were lower than those in OPN intervention group (all P<0.05); but there was no significant difference in the expression levels of HAS1, HAS2, HAS3 and HA between the CD44 homotype group and the OPN intervention group (all P>0.05). The results of OPN-CD44 axis study showed that: compared with the OA-NC group, the expression of CD44 in the rhOPN intervention group was slightly lower, but the protein and mRNA levels of HAS1 were significantly increased (all P<0.05); compared with the OA-NC group, the expression of CD44 was up-regulated, but the protein and mRNA level of HAS1 were significantly inhibited in the si-OPN intervention group (all P<0.05); compared with the OA-NC group, the protein and mRNA levels of HAS1 in the rhOPN+CD44 Ab group were also significantly inhibited (all P<0.05). CONCLUSIONS: The OPN in OA chondrocytes can promote the expression of HAS1, and the OPN can stimulate the secretion of HAS and induce HA expression by reacting with CD44 ligand receptor. They constitute the axis of OPN/CD44/HAS1, which plays an important role in regulating the expression of HA in chondrocytes.

Intestinal pseudo-obstruction as the initial manifestation of systemic lupus erythematosus.

Small bowel obstruction is common in emergency departments. However, the exact cause of intestinal pseudo-obstruction (IPO) is often misdiagnosed. IPO is considered a severe manifestation of systemic lupus erythematosus (SLE). However, IPO is rare as the initial manifestation of SLE. This paper reports a female patient who presented with IPO as the initial manifestation and was ultimately diagnosed with SLE. The 31-year-old female was definitively diagnosed with SLE after IPO symptoms for 1 month. She then presented multiple organ dysfunction syndrome (MODS) leading to a poor prognosis. Patients with unexplained SBO symptoms should be aware of systemic diseases. Early diagnosis and prompt medical treatment are crucial to avoid unnecessary surgery and obtain satisfactory outcomes.

[Levels of D-dimer, fibrinogen, and fibrinogen degradation product in patients with acute chest pain].

OBJECTIVE: To determine the relationship among the levels of D-dimer, fibrinogen (FIB), and fibrin degradation products (FDP) in acute fatal chest pain patients.
 Methods: We retrospectively analyzed the patients with aortic dissection (AD), pulmonary embolism (PE) or acute myocardial infarction (AMI) from May 1, 2017 to April 30, 2018. All the patients had a chest and/or back pain. Levels of D-dimer, FIB, and FDP were examined at the time of admission, and the patients were further diagnosed by computed tomography angiography (CTA) or percutaneous transluminal coronary intervention (PCI). The levels and negative rates of D-dimer, FIB, and FDP in patients with AD, PE, and AMI were compared.
 Results: A total of 234 patients were enrolled, including 95 AD, 98 AMI, and 41 PE. In the AD group, the AMI group and the PE group, the negative ratios of D-dimer were 13.68%, 70.41% and 4.88%, respectively; the negative ratios of FDP were 24.21%, 81.63% and 24.39%, respectively. There was no significant difference in negative rates of D-dimer and FDP between the AD group and the PE group (all P>0.05), but negative rates of D-dimer and FDP were significantly higher in the AMI group than those in the AD group and the PE group (all P<0.001). The level of D-dimer in the AMI group was significantly lower than that in the AD group or in the PE group (both P<0.001), while there was no statistically significant difference between the AD group and the PE group (P>0.05). However, there were no significant difference in the FIB levels among 3 groups (all P>0.05). The FDP level in the AMI group was significantly lower than that in the AD group or in the PE group (both P<0.001), while there was no statistically significant difference between the AD group and the PE group (P>0.05).
 Conclusion: The levels of D-dimer and FDP are increased in AD and PE patients and may be as the useful biomarkers for the high-risk chest pain patients but not for AMI.

Mitigation of reactive metabolite formation for a series of 3-amino-2-pyridone inhibitors of Bruton's tyrosine kinase (BTK).

Reactive metabolites have been putatively linked to many adverse drug reactions including idiosyncratic toxicities for a number of drugs with black box warnings or withdrawn from the market. Therefore, it is desirable to minimize the risk of reactive metabolite formation for lead molecules in optimization, in particular for non-life threatening chronic disease, to maximize benefit to risk ratio. This article describes our effort in addressing reactive metabolite issues for a series of 3-amino-2-pyridone inhibitors of BTK, e.g. compound 1 has a value of 459pmol/mg protein in the microsomal covalent binding assay. Parallel approaches were taken to successfully resolve the issues: establishment of a predictive screening assay with correlation association of covalent binding assay, identification of the origin of reactive metabolite formation using MS/MS analysis of HLM as well as isolation and characterization of GSH adducts. This ultimately led to the discovery of compound 7 (RN941) with significantly reduced covalent binding of 26pmol/mg protein.

Single nucleotide polymorphisms in the CD40 gene associate with the disease susceptibility and severity in knee osteoarthritis in the Chinese Han population: a case-control study.

BACKGROUND: This study explored the association between single nucleotide polymorphisms (SNPs) in the CD40 gene, rs4810485 G > T and rs1883832 C > T, as well as disease susceptibility and severity in knee osteoarthritis (KOA) in the Chinese Han population. METHOD: Peripheral venous blood was collected from 133 KOA patients (KOA group) and 143 healthy people (control group) from December 2012 to November 2013. The patients in the KOA group were classified into mild, moderate and severe groups according to disease severity. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to test the genotypes of all subjects. Binary logistic regression analyses were performed to analyze the risk factors for KOA. RESULTS: The KOA group was significantly different from the control group in living environment (P < 0.05). The KOA group had a lower frequency of TT genotype and T allele distribution of rs4810485 G > T compared with the control group, and rs4810485 G > T TT genotype and T allele may associate with low incidence of KOA (all P < 0.05). Besides, T allele and mutant homozygous TT genotype of rs1883832 C > T increased the susceptibility to KOA. Genotype and allele distribution of rs4810485 G > T and rs1883832 C > T were significantly different among the mild, moderate and severe groups (P < 0.05). There were more patients with rs4810485 G > T GG genotype and rs1883832 C > T TT genotype in the severe group than other genotypes of these two SNPs. According to binary logistic regression analysis, rs4810485 G > T TT genotype could alleviate disease severity in KOA, rs1883832 C > T TT genotype increase the severity of KOA and living environment is an important external factor that affects KOA severity. CONCLUSIONS: These data provide evidences that rs4810485 G > T and rs1883832 C > T in the CD40 gene may be associated with disease susceptibility and severity in KOA.

Role of integrins and their ligands in osteoarthritic cartilage.

Osteoarthritis (OA) is a degenerative disease, which is characterized by articular cartilage destruction, and mainly affects the older people. The extracellular matrix (ECM) provides a vital cellular environment, and interactions between the cell and ECM are important in regulating many biological processes, including cell growth, differentiation, and survival. However, the pathogenesis of this disease is not fully elucidated, and it cannot be cured totally. Integrins are one of the major receptors in chondrocytes. A number of studies confirmed that the chondrocytes express several integrins including α5ß1, αVß3, αVß5, α6ß1, α1ß1, α2ß1, α10ß1, and α3ß1, and some integrins ligands might act as the OA progression biomarkers. This review focuses on the functional role of integrins and their extracellular ligands in OA progression, especially OA cartilage. Clear understanding of the role of integrins and their ligands in OA cartilage may have impact on future development of successful therapeutic approaches to OA.

Role of osteopontin in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint swelling, joint tenderness, and destruction of synovial joints, leading to severe disability and premature mortality. RA is a multifactorial disease with genetic, environmental, and stochastic components related to its susceptibility. It has been demonstrated that the expression of osteopontin (OPN) is upregulated in the RA patients. Numerous studies have indicated that the full-length OPN or even OPN fragments, such as thrombin-cleaved OPN and its receptors, play the key roles in RA pathogenesis. Therapeutic application of siRNA to target OPN or neutralizing antibodies related to OPN epitopes in RA animal models are in progress, and some results are encouraging. However, there is a long way to go along with the clinical trials. This review focuses on the recent development in research associated with the OPN role in the pathogenesis of RA and provides insights concerning the OPN targeting as therapeutic approaches for patients with RA.

The Hippo-YAP Signaling Pathway in Osteoarthritis and Rheumatoid Arthritis.

Arthritis is the most prevalent joint disease and is characterized by articular cartilage degradation, synovial inflammation, and changes in periarticular and subchondral bone. Recent studies have reported that Yes-associated protein (YAP) and the transcriptional coactivator with PDZ-binding motif (TAZ) have significant effects on the proliferation, migration, and survival of chondrocytes and fibroblast-like synovial cells (FLSs). YAP/TAZ signaling pathway, as well as the related Hippo-YAP signaling pathway, are responsible for the condition of cells and articular cartilage in joints. They are tightly regulated to maintain metabolism in chondrocytes and FLSs because abnormal expression may result in cartilage damage. However, the roles and mechanisms of the Hippo-YAP pathway in arthritis remain largely unknown. This review summarizes the roles and key functions of YAP/TAZ and the Hippo-YAP signaling pathway in FLSs and chondrocytes for the induction of proliferation, migration, survival, and differentiation in rheumatoid arthritis (RA) and osteoarthritis (OA) research. We also discuss the therapeutic strategies involving YAP/TAZ and the related Hippo-YAP signaling pathway involved in OA.

A study on the safety and efficacy of endoscopic thyroidectomy via axillary approach for the treatment of thyroid cancer.

This study aims to evaluate the safety and efficacy of endoscopic thyroid cancer treatment using an axillary approach. Participants were allocated into 2 groups: one undergoing transaxillary endoscopic surgery and the other, traditional open surgery. We compared intraoperative and postoperative conditions, focusing on parameters such as intraoperative blood loss, duration of surgery, length of postoperative hospitalization, volume of postoperative drainage, number of lymph nodes cleared in the central region, neck pain scores, neck injury indices, cosmetic satisfaction, postoperative complications, and total hospitalization duration. Patients in the endoscopic treatment (ET) group experienced longer surgical times, less intraoperative bleeding, and increased postoperative drainage. These indicators showed significant differences between the groups (P < .05). For the group undergoing endoscopic surgery via the axillary approach, there was a lower neck pain score on the third postoperative day and higher cosmetic satisfaction at 3 months. However, there were no significant differences between the groups in terms of the number of lymph nodes cleared in the central area, and the incidence of complications such as difficulty breathing, difficulty swallowing, hoarseness, and subcutaneous hematoma (P > .05). The axillary approach endoscopic surgery group also showed significantly prolonged surgery times and postoperative hospital stays, with a significant increase in postoperative drainage fluid (P < .05). Concurrently, this technique involved smaller surgical incisions and effectively concealed scars in the armpit, leading to better outcomes in terms of intraoperative bleeding, neck pain scores, and postoperative cosmetic satisfaction. Non-inflatable ET via the axillary approach for treating thyroid cancer demonstrates promising efficacy and safety. It offers additional benefits of minimal pain and enhanced cosmetic outcomes, making it a viable option for clinical adoption and application.