GPR146 regulates pulmonary vascular remodeling by promoting pulmonary artery smooth muscle cell proliferation through 5-lipoxygenase.

The pathological feature of hypoxic pulmonary hypertension (PH) is pulmonary vascular remodeling (PVR), primarily attributed to the hyperproliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs). Existing PH-targeted drugs have difficulties in reversing PVR. Therefore, it is vital to discover a new regulatory mechanism for PVR and develop new targeted drugs. G protein-coupled receptor 146 (GPR146) is believed to participate in this process. This study aimed to investigate the role of GPR146 in PASMCs during PH. We investigated the role of GPR146 in PVR and its underlying mechanism using hypoxic PASMCs and mouse model (Sugen 5416 (20 mg/kg)/hypoxia). In our recent study, we have observed a significant increase in the expression of GPR146 protein in animal models of PH as well as in patients diagnosed with pulmonary arterial hypertension (PAH). Through immunohistochemistry, we found that GPR146 was mainly localized in the smooth muscle and endothelial layers of the pulmonary vasculature. GPR146 deficiency induction exhibited protective effects against hypoxia-induced elevation of right ventricular systolic blood pressure (RVSP), right ventricular hypertrophy, and pulmonary vascular remodeling in mice. In particular, the deletion of GPR146 attenuated the hypoxia-triggered proliferation of PASMCs. Furthermore, 5-lipoxygenase (5-LO) was related to PH development. Hypoxia and overexpression of GPR146 increased 5-LO expression, which was reversed through GPR146 knockdown or siRNA intervention. Our study discovered that GPR146 exhibited high expression in the pulmonary vessels of pulmonary hypertension. Subsequent research revealed that GPR146 played a crucial role in the development of hypoxic PH by promoting lipid peroxidation and 5-LO expression. In conclusion, GPR146 may regulate pulmonary vascular remodeling by promoting PASMCs proliferation through 5-LO, which presents a feasible target for PH prevention and treatment.

Case Series and a Literature Review: Two Ovarian Clear Cell Carcinoma Cases with Recurrent Endometriosis.

Introduction: Endometriosis-associated ovarian cancer (EAOC) is rare, occurring approximately in 1% of women with ovarian endometriosis. The main histological types are endometrioid adenocarcinoma and clear cell carcinoma (CCC), with the latter being the least common. Case Presentation: In our hospital, we recently summarized two patients with ovarian clear cell carcinoma with similar characteristics. They all had endometriosis for a long time and had undergone ovarian cyst removal due to a chocolate cyst. Unfortunately, the cyst recurred after surgery, and the histological diagnosis was clear cell carcinoma. In case 1, the expression of P53 was found in the tumor by regular examination, and the stage was IIB. In Case 2, we found it in intraoperative freezing; the stage was IA. Case 1 has been treated with the TP regimen six times, and the survival period has reached one year. Case 2 had a survival period of 6 years after completing six TP regimen treatments. Clinicians should pay attention to patients with a long history of endometriosis and postoperative recurrence of ovarian cysts accompanied by serum CA-125 of more than 200U/mL. Imaging examination has a good prospect in diagnosing malignant transformation of endometriosis, especially PET-CT. Conclusion: This case report highlights the risk factors related to the formation of ovarian clear cell carcinoma and suggests that the follow-up of patients with ovarian endometriosis is essential because of its recurrence characteristics. Radical surgery and postoperative platinum-containing chemotherapy are the primary treatment methods at present.

N6-Methyladenosine Modifications in Pulmonary Hypertension.

BACKGROUND: The most prevalent kind of RNA methylation modification existing in eukaryotes is N6-methyladenosine (m6A), which is a reversible type of posttranscriptional modification. SUMMARY: Many studies have reported that m6A participates in cell differentiation, self-renewal, invasion, and apoptosis by modifying protein synthesis. Furthermore, m6A modification is also involved in disease progression and pulmonary vascular remodeling in pulmonary hypertension. However, very few researchers have investigated the effect of m6A modifications on pulmonary hypertension. KEY MESSAGES: Here, we have reviewed the latest research advances in the field of m6A RNA methylation in pulmonary hypertension and explored its regulatory role in pulmonary hypertension development and progression.

Recent progress of nanomedicine in the treatment of Alzheimer's disease.

Alzheimer's disease (AD) is the most common cause of memory disruption in elderly subjects, with the prevalence continuing to rise mainly because of the aging world population. Unfortunately, no efficient therapy is currently available for the AD treatment, due to low drug potency and several challenges to delivery, including low bioavailability and the impediments of the blood-brain barrier. Recently, nanomedicine has gained considerable attention among researchers all over the world and shown promising developments in AD treatment. A wide range of nano-carriers, such as polymer nanoparticles, liposomes, solid lipid nanoparticles, dendritic nanoparticles, biomimetic nanoparticles, magnetic nanoparticles, etc., have been adapted to develop successful new treatment strategies. This review comprehensively summarizes the recent advances of different nanomedicine for their efficacy in pre-clinical studies. Finally, some insights and future research directions are proposed. This review can provide useful information to guide the future design and evaluation of nanomedicine in AD treatment.

Hypoxia activates GPR146 which participates in pulmonary vascular remodeling by promoting pyroptosis of pulmonary artery endothelial cells.

BACKGROUND: Hypoxia is a risk factor of pulmonary hypertension (PH) and may induce pulmonary artery endothelial cells (PAECs) injury and inflammation. Pyroptosis is a form of cell death through maturation and secretion of inflammatory mediators. However, the mechanistic association of pyroptosis, PAECs injury, and inflammation remain unknown. Here, we explored in detail the effects of hypoxia on pyroptosis of PAECs. EXPERIMENTAL APPROACH: Using RNA sequencing, we screened differentially expressed genes in pulmonary artery tissue of a Sugen5416/hypoxia-induced (SuHx) rat PH model. We examined the role of the differentially expressed gene G-protein coupled receptor 146 (GPR146) in PAECs through immunohistochemistry, immunofluorescence, CCK-8 assays, western blotings, real-time PCR, detection of reactive oxygen species, and lactate dehydrogenase release experiments. KEY RESULTS: According to RNA sequencing, GPR146 was 11.64-fold increased in the SuHx-induced PH model, compared to the controls. Further, GPR146 was highly expressed in pulmonary arterial hypertension human lung tissue and SuHx-induced rat PH lung tissues. Our results suggested that the expression of pyroptosis-related proteins was markedly increased under hypoxia, both in vivo and in vitro, which was inhibited by silencing GPR146. Moreover, inhibiting NLRP3 or caspase-1 effectively suppressed cleavage of caspase-1, production of interleukin (IL)-1ß, IL-6, and IL-18 in PAECs by hypoxia and overexpression of GPR146. CONCLUSION: Our results indicated that GPR146 induced pyroptosis and inflammatory responses through the NLRP3/caspase-1 signaling axis, thus triggering endothelial injury and vascular remodeling. Hypoxia may promote PAECs pyroptosis through upregulation of GPR146 and thereby facilitate the progression of PH. Taken together, these insights may help identify a novel target for the treatment of PH.

Effect of Myrcene on Th17/Treg Balance and Endocrine Function in Autoimmune Premature Ovarian Insufficiency Mice through the MAPK Signaling Pathway.

BACKGROUND: Premature ovarian insufficiency (POI) is a defect of ovarian functions in women younger than 40 years old. Although a large number of studies have focused on investigating autoimmune POI, its detailed pathogenesis is still largely unknown. Several studies have indicated that Myrcene exerted a part in the biological processes of various diseases. Nonetheless, whether Myrcene could influence the development of autoimmune POI remains to be elucidated. METHODS: POI model was established by injecting zona pellucida glycoprotein 3 (pZP3). Hematoxylin and eosin (H&E) staining was applied to evaluate the pathological features of ovarian tissues. Enzymelinked immunosorbent assay (ELISA) was used for assessing the concentrations of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH) and interleukin (IL)-17. Flow cytometry analysis was conducted for assessing the balance of Th17/Treg cells. RESULTS: The results showed that decreased levels of body weight, ovarian weight and ovarian index were reversed by Myrcene in POI model mice. The estrous cycles in mice were extended in pZP3 mice and Myrcene administration restored it to normal. The reduced number of primordial, primary, and secondary follicles as well as the increased number of atretic follicles in POI mice were offset by Myrcene administration. Moreover, Myrcene could modulate the Th17/Treg balance in autoimmune POI. Besides, Myrcene suppressed the MAPK signaling pathway in pZP3 mice. CONCLUSION: Myrcene regulated the Th17/Treg balance and endocrine function in autoimmune POI mice through the MAPK signaling pathway, which might provide a reference for improving the treatment of autoimmune POI.

The mechanism of the imbalance between proliferation and ferroptosis in pulmonary artery smooth muscle cells based on the activation of SLC7A11.

Pulmonary arterial hypertension (PAH) is a chronic, progressive pulmonary vascular disease. Pulmonary vascular remodelling (PVR) is one of the main pathological features of PAH. The main cause of PVR is cell death inhibition and excessive proliferation in pulmonary artery smooth muscle cells (PASMCs), which are also affected by oxidative stress. Ferroptosis is a newly identified form of cell death, which is associated with oxidative damage. It depends on the excessive accumulation of lipid peroxides and reactive oxygen species (ROS) in cells. Solute carrier family 7 member 11 (SLC7A11) is a subunit of the cystine/glutamate antiporter system Xc-, which inhibits ferroptosis by eliminating ROS through the promotion of GSH synthesis in cancer cells. However, very few studies exist on the relationship between ferroptosis and SLC7A11 in PAH. In this study, SLC7A11 was up-regulated in Sugen5416/hypoxia-induced PAH rats and patients with PAH. Moreover, SLC7A11 inhibited ferroptosis and promoted proliferation by overexpressing SLC7A11 in PASMCs. Additionally, ubiquitin aldehyde binding 1 (OTUB1), the main regulator of SLC7A11 stability, was involved in the ferroptosis and proliferation of PASMCs. Furthermore, erastin induced ferroptosis by inhibiting SLC7A11 and glutathione peroxidase 4 (GPX4) expressions in vivo and in vitro, suggesting that the continuous proliferation in hypoxic PASMCs could be reversed by erastin. Therefore, this study identifies novel targets and new research directions regarding PAH pathogenesis and treatment.

Rupture of ovarian endometriotic cyst complicated with endometriosis: A case report.

BACKGROUND: Endometriosis (EMs), an estrogen-dependent disease, refers to the appearance of mucosa-covered endometrial tissues (glandular and interstitial) growing in the uterine cavity outside the uterine myometrium. It is commonly seen in women aged 25 to 45, with an incidence of approximately 10%-15%. CASE SUMMARY: A 35-year-old unmarried female who denied a history of sex with an intact hymen had multiple dysmenorrhea and pain in the left lower abdomen that recurred during menstruation. Ultrasound examination revealed a dark cystic area measuring 4.9 cm × 4.6 cm on the left side with poor light transmittance, which suggested a left endometriotic cyst. The patient was treated with pain medications (four capsules t.i.d., p.o.). After one month, computed tomography of the abdomen and pelvis revealed a low-density focus measuring approximately 38 mm in diameter, a blurred mesentery fat plane in the pelvic cavity, and pelvic effusion. Ultrasound showed a complex echo density measuring 5.2 cm × 3.0 cm × 4.2 cm in the left ovarian area and a fluid sonolucent area with a depth of 2.0 cm in the pelvic cavity. Left ovarian cystectomy, electrocautery for endometriotic lesions, myomectomy, and pelvic adhesion lysis were performed under laparoscopy. The postoperative diagnosis was left ovarian chocolate cyst rupture and EMs (stage III, ovarian type, peritoneal type). CONCLUSION: Laparoscopic surgery can safely control the symptoms of EMs and effectively eradicate the disease.

The lncRNA PAHRF functions as a competing endogenous RNA to regulate MST1 expression by sponging miR-23a-3p in pulmonary arterial hypertension.

BACKGROUND: Emerging evidence has shown that long non-coding RNA (lncRNA) plays important roles in the development of pulmonary arterial hypertension (PAH). However, some new lncRNAs in patients with PAH are still lacking research. Herein, we examined the expression and role of lncRNA (pulmonary arterial hypertension related factor, PAHRF) in PAH. METHODS: LncRNA PAHRF expression and localization were analyzed by realtime PCR and fluorescence in situ hybridization. Proliferation and apoptosis were detected by MTT, CCK-8, EDU staining, JC-1 assay, flow cytometry and western blotting. Luciferase activity assay was used to identify PAHRF/ miR-23a-3p/serine/threonine kinase 4 (STK4/MST1) interaction. RESULTS: LncRNA PAHRF was down-regulated in both the PAs of PAH patients and hypoxic human pulmonary artery smooth muscle cells (PASMCs). The overexpression of PAHRF inhibited the proliferation and promoted the apoptosis of PASMCs. Similarly, we also found PAHRF overexpression decreased the proliferation under hypoxia condition. Knockdown of PAHRF exerted the opposite effects. Luciferase activity assay proved molecular binding between PAHRF and hsa-miR-23a-3p. Moreover, MST1 was confirmed to be the putative target gene and regulated by PAHRF/miR-23a-3p. In addition, we explored the molecular mechanism regulating the expression of miR-23a-3p, and found that lncRNA PAHRF acted as an endogenous sponge for miR-23a-3p, and silencing lncRNA PAHRF could up-regulate the expression of miR-23a-3p. On the contrary, PAHRF-overexpressing plasmid inhibited the expression of miR-23a-3p in hypoxia. CONCLUSIONS: Our present study reveals a novel PAH regulating model that is composed of PAHRF, miR-23a-3p, and MST1. The aim of this study is probably going to provide a new explanation and give a further understanding of the occurrence of vascular remodeling in PAH from the perspective competing endogenous RNA hypothesis.

Optimization of strontium aluminate-based mechanoluminescence materials for occlusal examination of artificial tooth.

This work presents a novel approach for evaluating the occlusal examination of artificial tooth based on the mechanoluminescence (ML) materials. The rare earth doped strontium aluminate (SrAl2O4: Eu2+, Dy3+; SAOED) was chosen as the ML material, which was further composited with the commercial denture base resin (DBR) to determine its feasibility for the mechanics analysis of artificial tooth occlusion. To eliminate negative factors for occlusal analysis, SAOED was first optimized to exhibit a rapid decay of afterglow and enhanced ML intensity. The luminescent characterizations of the SAOED/DBR composites suggest DBR is a desirable elastic-supporter for nondestructive ML generation. Furthermore, the introduction of SAOED improved the mechanical performance of DBR, and its biocompatibility was maintained at the same time. These results suggest the feasibility of the idea to detect the mechanics in occlusal examination of artificial tooth based on ML. The bright and sensitive ML from the constructed standard artificial tooth models could guide clinicians to purposefully adjust the occlusal surface until a balanced occlusion established.

Toxicity of porcelain-fused-to-metal substrate to zebrafish (Danio rerio) embryos and larvae.

AIMS: Porcelain-fused-to-metal (PFM) crowns are a standard restoration technique in dentistry, but toxicity of PFM in vivo has not been systematically evaluated. The present study evaluated the effects of various metal alloy shells of PFM crowns on the development of zebrafish embryos and larvae in order to determine the safety of these materials. MAIN METHODS: Gold palladium (Au-Pd), silver palladium (Ag-Pd), Nickel chromium (Ni-Cr), cobalt chromium (Co-Cr), titanium (Ti) alloy porcelain crowns were immersed in artificial saliva for 1, 4, and 7 weeks, and the leach solution was collected and used to treat zebrafish embryos at 4-144 h PFM. Toxicity was assessed based on mortality, spontaneous movement, heart rate, hatchability, malformation, and swimming behavior. KEY FINDINGS: The 1-week leachates of the five PFMs were not toxic to zebrafish. The rates of mortality and malformation of zebrafish in the Ni-Cr alloy group were increased whereas spontaneous movement, heart rate, and swimming behavior were decreased for 4- and 7-week leachates. SIGNIFICANCE: Among metal substrates commonly used in dental work, Ni-Cr alloy was most toxic, followed by Co-Cr and Ag-Pd alloys. Ti and Au-Pd alloys showed good biocompatibility and are therefore the most suitable materials for clinical applications.

Click hydrogels, microgels and nanogels: emerging platforms for drug delivery and tissue engineering.

Hydrogels, microgels and nanogels have emerged as versatile and viable platforms for sustained protein release, targeted drug delivery, and tissue engineering due to excellent biocompatibility, a microporous structure with tunable porosity and pore size, and dimensions spanning from human organs, cells to viruses. In the past decade, remarkable advances in hydrogels, microgels and nanogels have been achieved with click chemistry. It is a most promising strategy to prepare gels with varying dimensions owing to its high reactivity, superb selectivity, and mild reaction conditions. In particular, the recent development of copper-free click chemistry such as strain-promoted azide-alkyne cycloaddition, radical mediated thiol-ene chemistry, Diels-Alder reaction, tetrazole-alkene photo-click chemistry, and oxime reaction renders it possible to form hydrogels, microgels and nanogels without the use of potentially toxic catalysts or immunogenic enzymes that are commonly required. Notably, unlike other chemical approaches, click chemistry owing to its unique bioorthogonal feature does not interfere with encapsulated bioactives such as living cells, proteins and drugs and furthermore allows versatile preparation of micropatterned biomimetic hydrogels, functional microgels and nanogels. In this review, recent exciting developments in click hydrogels, microgels and nanogels, as well as their biomedical applications such as controlled protein and drug release, tissue engineering, and regenerative medicine are presented and discussed.

[Relationship between factors of labour pain and delivery outcomes].

OBJECTIVE: To evaluate factors associated with labor pain and delivery outcomes. METHODS: From Jul. to Dec. 2009, 111 normal singleton cephalic presentation pregnancies (including 5 elderly parturient) who delivered at the Department of Obstetrics and Gynecology, Second Affiliated Hospital, Zhejiang Chinese Medical University were enrolled in this study to evaluate the relationship between factors of labor pain and delivery outcomes. The labor pain of latent phase and active phase were scored by the visual analogue scale (VAS). Factors associated with pain included the age of parturient, the number of gravidity and parity, occupation, education profile, dwell location, etc. The questionnaire was designed by ourselves. Childbirth awareness, psychological preparation of delivery, emotional controllability, couple relationship, the relationship of parturient and mother-in-law, the relationship of parturient and parents, family economic status, use of sedative during the labor process and delivery outcomes were collected and analyzed. RESULTS: (1) Factors associated with pain: in the latent phase, the rate of moderate labour pain of 1/5 in women with more than 35 years old was statistically lower than 76.4% (81/106) in suitable age group (P < 0.05). The women with a good understanding about delivery had a statistically lower rate of moderate pain of 64.7% (44/68) than 88.4% (38/43) of those having a poor understanding (P < 0.05). The women who had a better couple relationship had a significantly higher rate of moderate pain of 77.2% (78/101) than 4/10 of those who had a general couple relationship (P < 0.05). There was significant difference in rate of moderate pain between pluripara group (50.0%, 11/22) and primipara group (79.8%, 71/89; P < 0.01). In the active phase, women with tense, scared or a poor emotion control expressed significantly severe labour pain (59.0%, 36/61) than 35.6% (16/45) in well-prepared group. The rate of severe labour pain in good control of emotion group of 44.8% (43/96) was a statistically lower than 9/10 in poor control group. There was a statistically lower severe labour pain in women given by sedatives (29.2%, 7/24) than 54.9% (45/82) in women without sedatives treatment (P < 0.05). (2) Delivery outcomes: in latent phase, the rates of fetal distress and cesarean section were 36.6% (30/82) and 39.0% (32/82) in moderate pain group, which were significantly higher than 13.8% (4/29) and 17.2% (5/29) in mild pain group. In active phase, the rate of fetal distress, cesarean section and postpartum hemorrhage were 36.5% (19/52), 40.4% (21/52) and 13.5% (7/52) in severe pain group, which were significantly higher than [18.5% (10/54); 20.4% (11/54); 0] in moderate pain group (P < 0.05). CONCLUSIONS: Women with poor understanding of delivery, tense, scared, poor emotion control, young age and uniparous have severe labour pain. Sedative use could alleviate pain in active phase. Women with mild labour pain have good delivery outcomes.

In-vivo studies on intraperitoneally administrated poly(vinyl alcohol).

The fate of poly(vinyl alcohol) (PVA, 195,000 g/mol) was studied in rabbits and nude mice after intraperitoneal (i.p.) administration. In-vivo fluorescence imaging using nude mice allowed for studies of tetramethylrhodamine labeled PVA distribution in the body and tracking the urinary excretion. The excreted PVA was studied in detail after collecting the urine of rabbits over a time period of 28 days. The PVA was separated from the urine by dialysis and analyzed by FTIR spectroscopy, (1)H-NMR spectroscopy, and size exclusion chromatography (SEC). Even after extensive dialysis, it was found that the excreted PVA showed a characteristic brownish color. The spectroscopic techniques revealed that this color was caused by the urine pigment (a metabolite of bilirubin) that could not be separated completely from the PVA. SEC showed unambiguously that the PVA with the very high molar mass had a glomerular permeability in the kidneys. Simultaneously, histological studies of the kidneys and the liver demonstrated that the tissues did not show any obvious damage.

[Pathogenesis and clinical significance of interleukin-8 and tumor necrosis factor-alpha in cervicitis].

OBJECTIVE: To investigate the relationship between local immune status of vagina and the occurrence of disease in patients with cervicitis. METHODS: ELISA were used to detect the level of interleukin (IL)-8 and tumor necrosis factor (TNF)alpha in vaginal douche of patients with cervicitis due to ureaplasma urealyticum, mycoplasma hominis, chlamydia trachomatis, neisseria gonorrhoeae and cervical erosion. RESULTS: Compared with the control group, the level of IL-8 in vaginal douche of patients with mycoplasma hominis cervicitis, chlamydia trachomatis cervicitis and neisseria gonorrhoeae cervicitis was significantly higher and there was significant difference ng/L: 371 +/- 34, 369 +/- 31, 339 +/- 36, vs 341 +/- 32, 338 +/- 33, 316 +/- 24, (all P < 0.01). Comparing the level of IL-8 in vaginal douche of patients with ureaplasma urealyticum cervicitis and cervical erosion with that of control group, there was no statistical difference (all P > 0.05). The level of TNF-alpha in vaginal douche of each group was remarkably higher than that of control group except for patients with cervical erosion. And statistically significant difference was found between them (all P < 0.01). CONCLUSION: With regards to the pathogenesis of cervicitis, local immune mechanism of vagina plays an important role in the occurrence of cervicitis. The role of IL-8 in pathogenesis of mycoplasma hominis cervicitis, chlamydia trachomatis cervicitis and neisseria gonorrhoeae cervicitis is likely to be more important.