NMT2 alleviates depression-like behavior in a rat model of chronic unpredictable stress: An integrated proteomic and phosphoproteomic analysis.

Proteomics has been widely used to investigate multiple diseases. Combining the analyses of proteomics with phosphoproteomics can be used to further explain the pathological mechanisms of depression. In this study, depression-like behavior was induced in a rat model of chronic unpredictable mild stress (CUMS). We subsequently conducted the sucrose preference test, open field experiment, and forced swimming test to assess depressive-like behavior. Proteomic and phosphoproteomic sequencing of the hippocampal tissues from depressive-like behavior and normal rats were analyzed to identify differentially expressed proteins (DEPs) and differentially phosphorylated proteins (DPPs). Differentially expressed phosphorylated proteins (DEPPs) were obtained by intersecting the DEPs and DPPs, and functional enrichment analysis, as well as ingenuity pathway analysis (IPA), were subsequently performed. The study also investigated correlations among the DEPPs and used qRT-PCR to quantify the expression levels of key genes. Five DEPPs were identified, Gys1, Nmt2, Lrp1, Bin1, and Atp1a1, which were found to activate the synaptogenesis signaling pathway, induce mitochondrial dysfunction, and activate the phosphoinositide biosynthesis and degradation pathways. The qRT-PCR results confirmed the proteomic findings for Gys1, Nmt2, Lrp1, and Atp1a1. Importantly, inhibiting Nmt2 was found to alleviate depression-like behavior and alleviate neuronal apoptosis in the hippocampus of CUMS rats. In conclusion, we identified five DEPPs associated with the synaptogenesis signaling pathway, mitochondrial dysfunction, and phosphoinositide biosynthesis and degradation in depression. Furthermore, NMT2 may be a potential target for the treatment or diagnosis of depression. Our findings provide novel insights into the molecular mechanisms of depression.

Protective role of TRPV2 in synaptic plasticity through the ERK1/2-CREB-BDNF pathway in chronic unpredictable mild stress rats.

PURPOSE: Chronic stress is a significant risk factor for mood disorders such as depression, where synaptic plasticity plays a central role in pathogenesis. Transient Receptor Potential Vanilloid Type-2 (TRPV2) Ion Channels are implicated in hypothalamic-pituitary-adrenal axis disorders. Previous proteomic analysis indicated a reduction in TRPV2 levels in the chronic unpredictable mild stress (CUMS) rat model, yet its role in synaptic plasticity during depression remains to be elucidated. This study aims to investigate TRPV2's role in depression and its underlying mechanisms. METHODS: In vivo and in vitro experiments were conducted using the TRPV2-specific agonist probenecid and ERK1/2 inhibitors SCH772984. In vivo, rats underwent six weeks of CUMS before probenecid administration. Depressive-like behaviors were assessed through behavioral tests. ELISA kits measured 5-HT, DA, NE levels in rat hippocampal tissues. Hippocampal morphology was examined via Nissl staining. In vitro, rat hippocampal neuron cell lines were treated with ERK1/2 inhibitors SCH772984 and probenecid. Western blot, immunofluorescence, immunohistochemical staining, and RT-qPCR assessed TRPV2 expression, neurogenesis-related proteins, synaptic markers, and ERK1/2-CREB-BDNF signaling proteins. RESULTS: Decreased hippocampal TRPV2 levels were observed in CUMS rats. Probenecid treatment mitigated depressive-like behavior and enhanced hippocampal 5-HT, NE, and DA levels in CUMS rats. TRPV2 activation countered CUMS-induced synaptic plasticity inhibition. Probenecid activated the ERK1/2-CREB-BDNF pathway, suggesting TRPV2's involvement in this pathway via ERK1/2. CONCLUSION: These findings indicate that TRPV2 activation offers protective effects against depressive-like behaviors and enhances hippocampal synaptic plasticity in CUMS rats via the ERK1/2-CREB-BDNF pathway. TRPV2 emerges as a potential therapeutic target for depression.

Blocking Two-Pore Domain Potassium Channel TREK-1 Inhibits the Activation of A1-Like Reactive Astrocyte Through the NF-κB Signaling Pathway in a Rat Model of Major Depressive Disorder.

Major depressive disorder (MDD) refers to a widespread psychiatric disorder. Astrocytes play a pivotal role in regulating inflammation which is a well-acknowledged key component in depression pathogenesis. However, the effects of the neuroinflammation-inducing A1-like astrocytes on MDD are still unknown. TWIK-related K+ channel 1 (TREK-1) has been demonstrated to regulate the action of antidepressants. Nevertheless, its mechanisms and effects on A1-like astrocyte stimulation in MDD are not clear. Therefore, we conducted in vivo and in vitro experiments using TREK-1 specific inhibitor spadin. In vivo, rats were subjected to a 6-week chronic unpredictable mild stress (CUMS) followed by spadin treatment. Behavioral tests were employed to surveil depressive-like behaviors. Hippocampal proteomic analysis was carried out with the purpose of identifying differentially expressed proteins after CUMS and spadin treatments. In vitro, astrocyte-conditioned medium and spadin were used to treat rat astrocyte cell line. The activated microglia, inflammatory factors, A1 astrocyte markers, and activated nuclear factor kappa B (NF-κB) pathway were later analyzed using immunofluorescence, western blot, and RT-qPCR. Our findings indicated that blockage of TREK-1 reduced CUMS-induced depressive-like behavior in rats, inhibited the microglial stimulation, reduced inflammatory factor levels, and suppressed the activation of A1-like reactive astrocytes in the hippocampus. We also verified that the suppression of A1-like astrocytes by spadin necessitated the NF-κB pathway. According to the findings, blocking TREK-1 inhibited the activation of A1-like reactive astrocytes via the NF-κB signaling pathway in MDD. Our study preliminarily identifies a novel antidepressant mechanism of TREK-1 action and provides a therapeutic path for MDD.

[Quality Control and Safety Evaluation of Culture Medium for Human Assisted Reproduction].

OBJECTIVE: In this paper, the key points of quality control and safety evaluation of human assisted reproductive medium were summarized to provide reference for the establishment of relevant standards and quality control in the future. METHODS: Through literature research, the key factors of quality control and risk control of human assisted reproductive medium were summarized, and the problems in clinical transformation were discussed. RESULTS: It is very important for the development of human assisted reproduction technology to study the active ingredients and their harmful degradation products and drugs in the culture medium of assisted reproduction. CONCLUSIONS: At present, the biggest challenge is to effectively control the quality of the culture medium for human assisted reproduction, establish corresponding inspection methods and quality standards for the key components, ensure the safety and effectiveness during the product shelf life, and thus improve the success rate of human assisted reproduction technology.

In vitro genotoxicity evaluation and metabolic study of residual glutaraldehyde in animal-derived biomaterials.

Glutaraldehyde (GA) is an important additive that is mainly used in animal-derived biomaterials to improve their mechanical and antimicrobial capacities. However, GA chemical toxicity and the metabolic mechanism remain relatively unknown. Therefore, residual GA has always been a major health risk consideration for animal-derived medical devices. In this study, extracts of three bio-patches were tested via the GA determination test and mouse lymphoma assay (MLA). The results showed that dissolved GA was a potential mutagen, which could induce significant cytotoxic and mutagenic effects in mouse lymphoma cells. These toxic reactions were relieved by the S9 metabolic activation (MA) system. Furthermore, we confirmed that GA concentration decreased and glutaric acid was generated during the catalytic process. We revealed GA could be oxidized via cytochrome P450 which was the main metabolic factor of S9. We found that even though GA was possibly responsible for positive reactions of animal-derived biomaterials' biocompatibility evaluation, it may not represent the real situation occurring in human bodies, owing to the presence of various detoxification mechanisms including the S9 system. Overall, in order to achieve a general balance between risk management and practical application, rational decisions based on comprehensive analyses must be considered.

Downregulation of miR-1224 protects against oxidative stress-induced acute liver injury by regulating hepatocyte growth factor.

OBJECTIVE: To study the effect of microRNA-1224 (miR-1224) on hydrogen peroxide (H2 O 2 )-induced oxidative stress injury in hepatocytes, and explore its underlying mechanism. METHODS: L02 cells were treated with H2 O 2 (100 mmol/L) to establish the model of an oxidative stress injury in hepatocytes. Quantitative reverse transcriptase polymerase chain reaction was used to detect the expression of miR-1224 and hepatocyte growth factor (HGF) in L02 cells. L02 cells were transfected with anti-miR-con (H 2 O 2 + anti-miR-con group), anti-miR-1224 (H 2 O 2 + anti-miR-1224 group), pcDNA3.1 (H 2 O 2 + ctrl group), pcDNA3.1-HGF (H 2 O 2 + HGF group), si-HGF and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + HGF group), si-NC and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + ctrl group) by liposome method. Cells without any treatment were regarded as a negative control (NC) group. The protein expression of HGF in each group cells was detected by Western blot analysis. Cell viability and apoptosis of each group were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay or flow cytometry, respectively. The interaction between miR-1224 and HGF was measured by dual luciferase reporter gene assay. RESULTS: The expression of miR-1224 was enhanced in H2 O 2 -treated L02 cells and its knockdown alleviated H 2 O 2 -induced suppression of viability and promotion of apoptosis. HGF is a target of miR-1224 and its overexpression abated H 2 O 2 -induced injury in hepatocytes. Moreover, silencing of HGF rescued the effect of downregulation of miR-1224 on cell viability and apoptosis in H 2 O 2 -treated L02 cell. CONCLUSION: Downregulation of miR-1224 could attenuate oxidative stress-induced inhibition of viability and increase of apoptosis in hepatocytes by targeting HGF, which may provide a target for potential therapy of acute liver injury.

Cook Cervical Ripening Balloon successfully prevents excessive hemorrhage combined with ultrasound-guided suction curettage in the treatment of cesarean scar pregnancy.

AIM: Cesarean scar pregnancy (CSP) is a rare occurrence of ectopic pregnancy. CSP incidence has increased significantly as a result of the increase in cesarean section rates. At present, there are no standard treatment guidelines for CSP; therefore, we report a minimally invasive treatment method for patients diagnosed with CSP. METHODS: This study included 15 women who were diagnosed with CSP. Ultrasound-guided suction curettage was performed on all patients. The Cook Cervical Ripening Balloon was used to tamponade and prevent hemorrhage during the procedure. In 12 patients, the balloon was placed immediately following ultrasound-guided suction curettage; in two patients, the balloon was placed when excessive bleeding occurred post-curettage; and in one patient, the balloon was placed after the gestational sac evacuated by itself, and then suction aspiration was performed on day 5, following the evacuation. Human chorionic gonadotropin levels were evaluated three days after the procedure. RESULTS: Placement and inflation of the Cook Cervical Ripening Balloon was well tolerated by all patients. The balloon tamponade effectively reduced or prevented vaginal bleeding in all patients, and none of the patients had an estimated blood loss higher than 1000 ml. CONCLUSIONS: Ultrasound-guided suction curettage is effective in the treatment of CSP. The Cook Cervical Ripening Balloon is easy to place and inflate and successfully prevented bleeding or assisted in the management of bleeding complications. We recommend the Cook Cervical Ripening Balloon as an adjuvant method for ultrasound-guided suction curettage for the treatment of CSP.

Temporarily Blocking the Uterine Artery to Dig Out a Diffused Adenomyosis Lesion Treated Laparoscopically.

STUDY OBJECTIVE: To show the tips and tricks of a simpler technique for temporary blocking of the uterine artery in laparoscopic resection of a diffuse adenomyosis lesion to make the procedure more efficient and reproducible. DESIGN: This study is designed to be a step-by-step explanation of the technique using videos and pictures (Canadian Task Force classification III). SETTING: Changzhou Maternal and Child Health Hospital, Changzhou, China. PATIENTS: Three patients (age 39-42 years, 41±1.73) were diagnosed with diffuse adenomyosis with severe secondary dysmenorrhea willing to reserve the uterus and a poor response to medical management. Gynecologic examination revealed that the uteri sizes were 9 to 14 weeks. Transvaginal ultrasonography revealed that the lesions were 4 to 7 cm in size. INTERVENTIONS: Laparoscopic resection of the diffuse adenomyosis lesion was conducted after temporary blocking of the uterine artery with a rubber belt. MEASUREMENTS AND MAIN RESULTS: The traditional methods for injecting diluted vasopressin in the myometrium around the affected area and a half-life period of 30 minutes were used. Many adenomyosis patients with severe dysmenorrhea and menometrorrhagia have a large lesion; thus, the operating time is longer. The maximum dose of vasopressin is 20 units daily, and hypertensive patients have a contraindication. We made an incision of the broad ligament of the avascular area near the uterine artery and pulled the rubber pressure pulse ligation tightly through to temporarily block the uterine artery without vasopressin completely through the laparoscopic resection of the diffuse adenomyosis lesion. Intraoperative blood loss was only 120 to 230 mL. In this video, we show a simpler technique for temporarily blocking the uterine artery in laparoscopic resection of diffuse adenomyosis with a stepwise expiation. A levonorgestrel-releasing intrauterine system (Mirena; Bayer, Leverkusen, Germany) was placed in the uterus from the vagina immediately after surgery. At the 3-25 month (10.67±12.42) follow-up, visual analog scale scores were obviously reduced, and the menstrual quantity and amenorrhea dramatically declined after the surgery. All patients had no recurrence and no Mirena loss as assessed by vaginal ultrasound and the visual analog scale [1]. Estrogen was maintained at the normal level after 3 months. This study was approved by the Institutional Review Board of the Changzhou Maternal and Child Health Hospital Affiliated to Nanjing Medical University. CONCLUSION: The incidence of adenomyosis is a newer trend that is being used in patients with a poor response to medical management of uterine lesions and large lesions in China. Using the rubber belt to temporarily block the uterine artery in laparoscopic resection of the diffuse adenomyosis lesion offers the possibility of the rubber belt being effective, safe, and reproducible. Minimally invasive surgery in expert hands is the preferred solution of an increasing number of patients after drug treatment failure.

LTPB2 acts as a prognostic factor and promotes progression of cervical adenocarcinoma.

Latent transforming growth factor-beta-1 binding protein-2 (LTBP-2) is a member of the fibrillin/LTBP super family of extracellular matrix proteins, found to be overexpressed in certain malignant tumors. However, the clinical significance and biological role of LTBP-2 in cervical adenocarcinoma has remains unclear. We found that the expression of LTBP2 was higher in cervical adenocarcinoma than in normal cervical epithelial tissue as assessed by immunohistochemistry. Expression of LTBP2 is related to clinical stage, cervical tumor size, depth of cervical stromal invasion and lymph node metastasis. Knockdown of LTBP2 expression can inhibit the proliferation and migration of HeLa cells. Moreover, LTBP2 knockdown affected multiple tumor-related pathway genes including: the MAPK signaling pathway, the PI3K-AKT signaling pathway, receptor tyrosine kinase signaling and the P53 pathway. Taken together, this work suggests that LTBP2 may promote the development of cervical adenocarcinoma and serve as a prognostic factor in the clinical evaluation of patients with cervical adenocarcinoma. Our findings provide a new strategy for the diagnosis and treatment of cervical adenocarcinoma.