Shikonin reduces M2 macrophage population in ovarian cancer by repressing exosome production and the exosomal galectin 3-mediated ß-catenin activation.

BACKGROUND: Shikonin (SK), a naphthoquinone with anti-tumor effects, has been found to decrease production of tumor-associated exosomes (exo). This study aims to verify the treatment effect of SK on ovarian cancer (OC) cells, especially on the production of exo and their subsequent effect on macrophage polarization. METHODS: OC cells SKOV3 and A2780 were treated with SK. The exo were isolated from OC cells with or without SK treatment, termed OC exo and SK OC exo, respectively. These exo were used to treat PMA-induced THP-1 cells (M0 macrophages). M2 polarization of macrophages was determined by measuring the M2 specific cell surface markers CD163 and CD206 as well as the secretion of M2 cytokine IL-10. The functions of galectin 3 (LGALS3/GAL3) and ß-catenin in macrophage polarization were determined by gain- or loss-of-function assays. CB-17 SCID mice were subcutaneously injected with SKOV3 cells to generate xenograft tumors, followed by OC exo or SK OC exo treatment for in vivo experiments. RESULTS: SK suppressed viability, migration and invasion, and apoptosis resistance of OC cells in vitro. Compared to OC exo, SK OC exo reduced the M2 polarization of macrophages. Regarding the mechanism, SK reduced exo production in cancer cells, and it decreased the protein level of GAL3 in exo and recipient macrophages, leading to decreased ß-catenin activation. M2 polarization of macrophages was restored by LGALS3 overexpression but decreased again by the ß-catenin inhibitor FH535. Compared to OC exo, the SK OC exo treatment reduced the xenograft tumor growth in mice, and it decreased the M2 macrophage infiltration within tumor tissues. CONCLUSION: This study suggests that SK reduces M2 macrophage population in OC by repressing exo production and blocking exosomal GAL3-mediated ß-catenin activation.

Age, growth, reproduction and status of resource development of Ptychidio jordani, a critically endangered freshwater fish in the Hongshui River, China.

The age, growth, reproduction and resource development status of Ptychidio jordani, as a critically endangered freshwater fish in the Hongshui River, China, was studied in this work. A total of 525 specimens were collected monthly using the cages and gillnets from October 2021 to September 2022 in the Hongshui River. The scale was used for age determination, and the maximum age for both female and male was estimated to be 5 years and 3 years, respectively. Female and male P. jordani showed different growth patterns, which were expressed as Lt  = 261.3 (1-e-0.4885(t-0.1476) ) and Lt  = 251.2 (1-e-0.4758(t+0.9643) ), respectively. The overall sex ratio was 1:0.47 (female:male). Female attained sex maturity at 2.34 years (192 mm body length). Month variation of the gonad somatic index indicated that the spawning period occurred from April to October. The absolute fecundity was estimated at 9046 ± 3434 eggs per individual, and the relative fecundity was 38.08 ± 15.77 eggs per gram. The exploitation rate of female and male was 0.233 and 0.495, which indicated that P. jordani was not overfishing. This study provided data on the key life-history traits of P. jordani, which has not been known previously and is essential for conservation strategy and policy development.

Blood and CSF chemokines in Alzheimer's disease and mild cognitive impairment: a systematic review and meta-analysis.

OBJECTIVE: Chemokines, which are chemotactic inflammatory mediators involved in controlling the migration and residence of all immune cells, are closely associated with brain inflammation, recognized as one of the potential processes/mechanisms associated with cognitive impairment. We aim to determine the chemokines which are significantly altered in Alzheimer's disease (AD) and mild cognitive impairment (MCI), as well as the respective effect sizes, by performing a meta-analysis of chemokines in cerebrospinal fluid (CSF) and blood (plasma or serum). METHODS: We searched three databases (Pubmed, EMBASE and Cochrane library) for studies regarding chemokines. The three pairwise comparisons were as follows: AD vs HC, MCI vs healthy controls (HC), and AD vs MCI. The fold-change was calculated using the ratio of mean (RoM) chemokine concentration for every study. Subgroup analyses were performed for exploring the source of heterogeneity. RESULTS: Of 2338 records identified from the databases, 61 articles comprising a total of 3937 patients with AD, 1459 with MCI, and 4434 healthy controls were included. The following chemokines were strongly associated with AD compared with HC: blood CXCL10 (RoM, 1.92, p = 0.039), blood CXCL9 (RoM, 1.78, p < 0.001), blood CCL27 (RoM, 1.34, p < 0.001), blood CCL15 (RoM, 1.29, p = 0.003), as well as CSF CCL2 (RoM, 1.19, p < 0.001). In the comparison of AD with MCI, there was significance for blood CXCL9 (RoM, 2.29, p < 0.001), blood CX3CL1 (RoM, 0.77, p = 0.017), and blood CCL1 (RoM, 1.37, p < 0.001). Of the chemokines tested, blood CX3CL1 (RoM, 2.02, p < 0.001) and CSF CCL2 (RoM, 1.16, p = 0.004) were significant for the comparison of MCI with healthy controls. CONCLUSIONS: Chemokines CCL1, CCL2, CCL15, CCL27, CXCL9, CXCL10, and CX3CL1 might be most promising to serve as key molecular markers of cognitive impairment, although more cohort studies with larger populations are needed.

Blood and CSF Homocysteine Levels in Alzheimer's Disease: A Meta-Analysis and Meta-Regression of Case-Control Studies.

Objective: Hyperhomocysteinemia (HHcy), as an important risk factor for Alzheimer's disease (AD), would aggravate cognitive dysfunction. This study aimed to investigate whether and to what degree the homocysteine (Hcy) levels in blood and cerebrospinal fluid (CSF) were elevated in AD patients compared with healthy controls and to explore the factors related to the elevated Hcy levels in AD patients. Methods: PubMed and Embase databases were searched to identify eligible studies, and study quality was evaluated using the Newcastle-Ottawa Quality Assessment Scale. Ratio of mean (RoM) Hcy concentrations was used as a measure of fold-change between AD patients and healthy control subjects. Results: We identified 35 eligible studies, consisting a total of 2172 patients with AD and 2289 healthy controls. The pooled results showed that patients with AD had a significantly higher blood level of Hcy (RoM, 1.32; 95% CI, 1.25-1.40; p<0.001) than controls did, with large heterogeneity across studies (I2=81.4%, p<0.001). Hcy level in CSF did not differ significantly between patients with AD than controls (RoM, 1.12; 95% CI, 0.90-1.39, p=0.293; I2=69.4%, p=0.02). A random effects meta-regression analysis revealed that there was an inverse correlation between the blood levels of Hcy and folate (p=0.006). There was no link found between the blood levels of vitamin B12, or the Mini-Mental Status Examination scores reflecting the degree of cognitive impairment, and blood levels of Hcy. Conclusion: Regardless of dementia severity, there is an approximate one-third increase in blood Hcy in AD patients, which is robustly associated with a decreased level of blood folate in AD, but not with that of blood vitamin B12 nor the degree of dementia. Future investigation on the cause-and-effect link between Hcy and folate is warranted to clarify this issue.

Diversity, Distribution, and Biogeography of Freshwater Fishes in Guangxi, China.

The Guangxi Zhuang Autonomous Region has the largest number of cavefish species in the world and is a global biodiversity hotspot. In this study, a species list of freshwater fishes in 12 Sub-basins of Guangxi was compiled systematically. Moreover, the species composition and distribution of the diversity were analyzed via the G-F index, taxonomic diversity index, and beta diversity index. Results showed that 380 species of freshwater fishes were recorded in this region, which belonged to 158 genera in 43 families and 17 orders in 2 phyla, in which 128 species of endemic fishes and 83 species of cavefish accounted for 33.68% and 21.84%, respectively. The species diversity from northwest to southeast gradually decreased for most Sub-basins. The G-F index has generally risen in recent years. The taxonomic diversity index showed that the freshwater fish taxonomic composition in Guangxi is uneven. The spatial turnover component was the main contributor to beta diversity. A cluster analysis showed that the 12 Sub-basins in the study area could be divided into four groups, and the phylogenetic relationships of freshwater fishes in Guangxi generally reflect the connections between water systems and geological history. The freshwater fish system in Guangxi, which belonged to the South China division in the Southeast Asiatic subregion of the Oriental region, originated in the early Tertiary period. The results will provide the information needed for freshwater fish resource protection in Guangxi and a reference for promoting the normalization of fish diversity conservation in the Pearl River Basin and other basins.

Collagen type X alpha 1 promotes proliferation, invasion and epithelial-mesenchymal transition of cervical cancer through activation of TGF-ß/Smad signaling.

Background: Collagen type X alpha 1 (COL10A1) belongs to the collagen family and constitutes the main component of the interstitial matrix. COL10A1 was found to be dysregulated in various cancers, and to participate in tumorigenesis. However, the role of COL10A1 in cervical cancer (CC) remains unclear. Methods: Expression of COL10A1 in CC cells and tissues was detected by western blot and qRT-PCR. CC cells were transfected with pcDNA-COL10A1 or si-COL10A1, and the effect of COL10A1 on cell proliferation of CC was assessed by MTT and colony formation assays. Cell metastasis was detected by wound healing and transwell assays. Western blot was applied to evaluate epithelial-mesenchymal transition. Results: COL10A1 was significantly elevated in CC tissues and cells (P < 0.001). Over-expression of COL10A1 increased cell viability of CC (P < 0.001), and enhanced the number of colonies (P < 0.001). However, knockdown of COL10A1 reduced the cell proliferation of CC (P < 0.001). Over-expression of COL10A1 also promoted cell migration (P < 0.001) and invasion (P < 0.001) of CC, whereas silencing of COL10A1 suppressed cell metastasis (P < 0.001). Protein level of E-cadherin in CC was reduced (P < 0.05), whereas N-cadherin and vimentin were enhanced by COL10A1 over-expression (P < 0.001). Silencing of COL10A1 reduced the protein level of TGF-ß1 (P < 0.01), and down-regulated the phosphorylation of Smad2 and Smad3 in CC (P < 0.001). Conclusion: Down-regulation of COL10A1 suppressed cell proliferation, metastasis, and epithelial-mesenchymal transition of CC through inactivation of TGF-ß/Smad signaling.

CAPN6 Regulates Uterine Leiomyoma Cell Proliferation and Apoptosis through the Rac1-Dependent Signaling Pathway.

OBJECTIVE: Calpain 6 (CAPN6) is one of the calcium-dependent intracellular nonlysosomal proteases that are dysregulated in uterine leiomyomas (UtLMs). However, its function and mechanism in UtLMs is still unknown. METHODS: The correlation between CAPN6 expression and UtLMs was analyzed by the Gene Expression Omnibus (GEO). The expression of CAPN6 and Rac1 was detected by quantitative real-time PCR (qPCR) and western blot analysis. Cell proliferation ability was analyzed by a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis was detected by flow cytometry. RESULTS: CAPN6 was overexpressed in UtLMs compared with uterine smooth muscle cells (UtSMCs). The downregulation of CAPN6 resulted in decreased cell proliferation and increased apoptosis of UtLMs. Furthermore, mechanical investigations revealed that these inhibitory effects were correlated with Rac1/PAK1 signaling pathways. Silencing the expression of CAPN6 resulted in decreased Rac1 and phospho-PAK1. On the other hand, upregulated Rac1 expression could reverse the reduced phosphorylation of PAK1 induced by CAPN6 silencing. CONCLUSIONS: This data suggests that CAPN6 regulates UtLMs proliferation and apoptosis while being mediated through the Rac1/PAK1 signaling pathway.

A randomized, controlled study comparing minilaparotomy versus isobaric gasless laparoscopic assisted minilaparotomy myomectomy for removal of large uterine myomas: short-term outcomes.

OBJECTIVE: To compare the operative data and early postoperative outcomes for myomectomy performed by minilaparotomy (MLT) with isobaric laparoscopic assisted minilaparotomy myomectomy (LM) in a series of patients with large uterine myomas (>or=5 cm) randomly assigned to each surgical technique. STUDY DESIGN: 80 patients were randomized blindly using a computer randomization list to MLT (n=40) or LM (n=40). RESULTS: The mean (+/-SD) operating time was significantly shorter after LM than after MLT (75.50+/-25.70 vs 96.00+/-26.20 min; p<0.01). Intraoperative blood loss was less with LM (72.15+/-44.00 vs 96.21+/-38.50 ml; p<0.05), and DeltaHb was less with LM (1.21+/-0.55 vs 1.64+/-0.57; p<0.05). No intraoperative complications occurred, and no case was returned to the theater in either group. No conversion to standard laparotomy was necessary. Hospitalization was shorter after LM than after MLT (4.30+/-1.20 vs 6.90+/-2.70 days; p<0.01). Postoperative ileus was shorter after LM than after MLT (26.20+/-4.20 vs 40.50+/-4.90h; p<0.01). The mean VAS score at 12h for abdominal pain was 5.5+/-0.7 in the LM group and 5.2+/-0.8 in MLT group (p<0.05), whereas it was analogous in the two groups at 24h, and at 48h was 3.4+/-1.1 in the LM group and 4.2+/-1.1 in the MLT group (p<0.05), and no difference between two groups was detected in the overall mean (at 12, 24 and 48h). CONCLUSIONS: Several surgical and immediate postoperative outcomes were significantly better in the LM group than in the MLT group.

A randomized trial of laparoscopic versus laparoscopic-assisted minilaparotomy myomectomy for removal of large uterine myoma: short-term outcomes.

STUDY OBJECTIVE: To compare operative data and early postoperative outcomes for myomectomy performed by isobaric gasless laparoscopic-assisted minilaparotomy (LA-MLT) compared with those by isobaric gasless laparoscopy (LA) in a series of patients with large uterine leiomyomas (> or =5 cm) randomly assigned to each surgical technique. DESIGN: Randomized trial (Canadian Task Force classification I). SETTING: University departments of gynecology in Jiangyin and Nanjing, Jiangsu Province, Republic of China. PATIENTS: Fifty-two patients were randomized blindly by use of a computer randomization list to either LA (n = 26) or LA-MLT (n = 26). MEASUREMENTS AND MAIN RESULTS: The mean operating time was significantly shorter after LA-MLT than after LA (75.50 +/- 25.70 vs 96.00 +/- 26.20 minutes); the 95% confidence interval (95% CI) was 20.5 (6.04-34.96; p =.006). The intraoperative blood loss was less with LA-MLT (71.92 +/- 18.98 vs 96.34 +/- 32.42 mL); the 95% CI was 24.42 (9.63-39.22; p =.002); and the hemoglobin level decrease was less with LA-MLT (1.22 +/- 0.61 vs 1.65 +/- 0.61); the 95% CI was 0.43 (0.09-0.76; p =.014). There was a difference of the visual analog scale score among the 3 time points (0, 12, and 24 hours) with the 2 groups combined (F = 844.15, p <.001); and no difference in the visual analog scale score between the treatment groups, with values at all time points averaged and over time (p >.05). With regard to the early postoperative outcome, no difference between the 2 groups was detected in hospitalization days (1.81 +/- 0.57 vs 2.04 +/- 0.66 days; 95% CI 20.23 [20.57-0.11, p =.183]); and postoperative ileus (23.20 +/- 4.37 vs 22.80 +/- 3.94; 95% CI 0.39 [21.93-2.70, p =.738]). CONCLUSIONS: Several surgical and immediate postoperative outcomes were significantly better in the gasless LA-MLT group than in the LA group.