[Establishment and preliminary application of organoids in ovarian cancer].

Objective: To explore the establishment and application of ovarian cancer organoids. Methods: Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 µmol/L (5, 10, 20, 40, 80 µmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC50) in different organoids. Results: (1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC50 of (29.5±15.8) µmol/L. Moreover, IC50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) µmol/L vs (24.0±12.1) µmol/L; t=3.429, P=0.022]. Conclusions: Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

[Beneficial effects of Schisandrin B on the cardiac structure and function in a mice model of myocardial infarction].

Objective: To investigate whether Schisandrin B (Sch B) could improve cardiac structure and function in myocardial infarction (MI) mice and related mechanisms. Methods: Male C57BL/6J mice were randomized into sham (n=8), MI+ Sch B (n=24, 80 mg·kg(-1)·d(-1) per gavage) or MI+ vehicle (n=24, equal volume). After treatment for 3 weeks, cardiac function was detected by echocardiography measurement.Infarction size was measured by Evans blue and TTC staining.HE and Masson trichrome staining were used to observe the myocardial inflammation, structure and fibrosis.TNF-α, TGF-ß, IL-1ß were detected by ELISA. The apoptosis index of ischemic myocardial cells was detected by immunofluorescence. NF-κB, Bcl-2, Bax, Akt phosphorylated Akt(p-Akt), eNOS, phosphorylated eNOS (p-eNOS) were detected by Western blot. Results: Three weeks after operation, survival rate (83.33% vs. 54.17%, P<0.05), LVEF((51.77±8.50)% vs.(40.23±8.30)%, P<0.05), LVFS((26.44±5.15)% vs. (19.53±4.56)%, P<0.05)were significantly higher; LVEDD ((4.13±0.40) mm vs.(4.44±0.46)mm, P<0.05), LVESD((3.07±0.39) mm vs. (3.46±0.52)mm, P<0.05), the heart weight/body weight ratio((0.59±0.06)% vs. (0.68±0.10)%, P<0.05)was significantly lower and infarct size ((23.4±2.36)% vs. (39.4±2.06)%, P<0.05) was significantly reduced in MI+ Sch B group than those in MI+ vehicle group. In MI+ vehicle group, HE staining showed a large number of inflammatory cells in the peri-infarctl region, and the permutation structure was very disorganized, while above changes were significantly reduced in the MI+ Sch B group. Masson staining results showed that the degree of myocardial fibrosis in MI+ Sch B group was significantly less than that of MI+ vehicle group.Moreover, Sch B could down-regulate some inflammatory cytokines, like NF-κB、TGF-ß、TNF-α and IL-1ß, activate Akt-eNOS pathway, upgrade Bcl-2 and downgrade Bax and reduce cell apoptosis as compared with MI+ vehicle group (all P<0.05). Conclusions: Our results demonstrate that Sch B can reduce inflammation, inhibit apoptosis, and attenuate cardiac remodeling and improve cardiac function in this mice MI model.Sch B might serve as a potential novel therapeutic agent for ischemic heart disease.

Bone-cartilage interface crosstalk in osteoarthritis: potential pathways and future therapeutic strategies.

Currently, osteoarthritis (OA) is considered a disease of the entire joint, which is not simply a process of wear and tear but rather abnormal remodelling and joint failure of an organ. The bone-cartilage interface is therefore a functioning synergistic unit, with a close physical association between subchondral bone and cartilage suggesting the existence of biochemical and molecular crosstalk across the OA interface. The crosstalk at the bone-cartilage interface may be elevated in OA in vivo and in vitro. Increased vascularisation and formation of microcracks associated with abnormal bone remodelling in joints during OA facilitate molecular transport from cartilage to bone and vice versa. Recent reports suggest that several critical signalling pathways and biological factors are key regulators and activate cellular and molecular processes in crosstalk among joint compartments. Therapeutic interventions including angiogenesis inhibitors, agonists/antagonists of molecules and drugs targeting bone remodelling are potential candidates for this interaction. This review summarised the premise for the presence of crosstalk in bone-cartilage interface as well as the current knowledge of the major signalling pathways and molecular interactions that regulate OA progression. A better understanding of crosstalk in bone-cartilage interface may lead to development of more effective strategies for treating OA patients.

Basic science and clinical application of platelet-rich plasma for cartilage defects and osteoarthritis: a review.

Cartilage defects (CDs) and the most common joint disease, osteoarthritis (OA), are characterized by degeneration of the articular cartilage that ultimately leads to joint destruction. Current treatment strategies are inadequate: none results in restoration of fully functional hyaline cartilage, for uncertain long-term prognosis. Tissue engineering of cartilage with auto-cartilage cells or appropriate mesenchymal stem cell (MSC)-derived cartilage cells is currently being investigated to search for new therapies. Platelet-rich plasma (PRP), an autologous source of factors obtained by centrifugation, possesses various functions. For culture of MSCs and cartilage cells, it might be substituted for fetal bovine serum (FBS) with high efficiency and safety. It enhances the regeneration of cartilage cells when added to cartilage tissue engineering constructs for repairing CDs and as regenerative injection therapy for OA. But challenges also remain. Some of the growth factors (GFs) present in PRP have negative effects on the OA joint. It is therefore unlikely that a mix of GFs some of which have negative effects in the OA joint, as present in PRP, will be of benefit in OA. Future directions of PRP application may concentrate on seeking an appropriate and innocuous agent like anti-VEGF antibody that can modulate and control the effect of PRP.

PAX5-induced upregulation of LINC01194 exerts oncogenic properties by regulating GOLPH3 expression via miR-486-5p in prostate cancer.

OBJECTIVE: Several studies have demonstrated that long non-coding RNA can act as crucial roles during the progression of various tumors, including prostate cancer (PCa). We aimed to determine lncRNA LINC01194(LINC01194) expression in prostate cancer (PCa) and examine its influence on tumor behaviors of PCa cells. PATIENTS AND METHODS: RT-PCR was performed to examine LINC01194 and PAX5's expression levels in PCa tissues and cell lines. Luciferase reporter and chromatin immunoprecipitation (ChIP) assays were performed to explore whether PAX5 could activate the transcription of LINC01194. Cell viability, migration and invasion were assessed by CCK-8, colony formation, transwell assay and Wound-healing assays. Bioinformatics and Dual-Luciferase assays were used to investigate the interaction between LINC01194 and miR-486-5p, as well as between miR-486-5p and GOLPH3. Western blot was applied for detecting the expressions of the related proteins. RESULTS: LINC01194 was highly expressed in PCa specimens and cell lines. PAX5 could bind directly to LINC01194 promoter region and activate its transcription. Functionally, the proliferation and metastasis of PCa cells were substantially impeded by LINC01194 silencing in vitro and in vivo. Mechanistically, LINC01194 promoted PCa progression by serving as a sponge of miR-486-5p to increase GOLPH3 expression. CONCLUSIONS: Our study identifies LINC01194 as a tumor promotor in PCa and implicates the LINC01194/miR-486-5p/GOLPH3 axis in the PCa progression.

[Efficacy and safety of intranasal dexmedetomidine premedication for children undergoing CT or magnetic resonance imaging: a systematic review and meta-analysis].

Objective: To review the efficacy and safety in children receiving intranasal dexmedetomidine premedication before CT or magnetic resonance imaging (MRI). Methods: A literature search (search terms included "dexmedetomidine" "intranasal drug administration" "children" "CT" and "MRI") was conducted using Embase, PubMed, the Cochrane Library, ClinicalTrials.gov, CNKI, Wanfang, VIP database and Chinese Biomedical Literature Database (searched from inception to March 18, 2019). Randomized controlled trials of children receiving intranasal dexmedetomidine versus chloral hydrate, ketamine or midazolam premedication before CT or MRI were included. The Cochrane Reviewers' Handbook 5.1.0 was used to evaluate the quality of the enrolled studies. The primary outcomes were sedation success rate and sedation induction time. The secondary outcomes included respiratory depression, heart rate, systolic blood pressure and blood oxygen saturation. Statistical analyses were performed using the Review Manager 5.3 software. Results: A total of 1 167 participants in 9 randomized controlled trials were included. The results of meta-analysis showed that intranasal dexmedetomidine premedication provided higher sedation success rate than oral chloral hydrate (relative risk (RR) =1.13, 95% confidence interval (CI) 1.02 to 1.26, P=0.020). There was no significant difference between intranasal dexmedetomidine and midazolam. In addition, the sedation induction time of intranasal dexmedetomidine group was significantly shorter than that in the oral chloral hydrate group (weighted mean difference -1.49, 95% CI -2.87 to -0.11; P=0.030), but showed no significant difference as compared with that of intranasal ketamine or midazolam. The patients treated with intranasal dexmedetomidine also showed significantly lower heart rate (RR=4.78, 95%CI 1.85-12.35, P=0.001) and less respiratory depression (RR=0.28, 95%CI 0.09-0.87, P=0.030). There were no intergroup differences in systolic blood pressure and blood oxygen saturation. Conclusions: Intranasal dexmedetomidine provided more effective sedation and higher safety in children undergoing CT or MRI. As this meta-analysis is limited by the small sample size, further high-quality randomized controlled trials are needed.

Protective effects of sodium p-aminosalicylic acid on learning and memory via increasing the number of basal forebrain choline acetyltransferase neurons in manganese-exposed rats.

This study was conducted to investigate the protective effects of sodium p-aminosalicylic acid (PAS-Na) on learning and memory via increasing the number of basal forebrain choline acetyltransferase (ChAT) neurons in manganese (Mn)-exposed rats. Male Sprague Dawley rats were divided into following groups: the normal control I, II, and III groups, the model I, II, and III groups, low- and high-dose PAS-Na treatment (L- and H-PAS) group, PAS-Na prevention (PAS-P) group, and PAS-Na treatment (PAS-T) group. The model I, II, and III groups, L- and H-PAS, and PAS-T groups received intraperitoneal (i.p.) injection of 15 mg/kg manganese chloride tetrahydrate (MnCl2·4H2O) for 3 or 12 weeks, while the normal control I, II, and III groups received i.p. injection of an equal volume of saline; L- and H-PAS and PAS-T groups received back subcutaneous (s.c.) injection of PAS-Na (100 and 200 mg/kg) for the next 5 or 6 weeks, whereas model I and II group received back s.c. injection of an equal volume of saline. However, PAS-P group received back s.c. injection of 200 mg/kg PAS-Na + i.p. injection of 15 mg/kg MnCl2·4H2O for 12 weeks. Mn exposure significantly reduced the ability of spatial learning and memory capability, while PAS-Na prevention recovered it. Mn decreased the number of ChAT-positive neurons in vertical limb nucleus of the basal forebrain diagonal band/horizontal limb nucleus of the basal forebrain diagonal band and ChAT protein activity and treatment or prevention with PAS-Na restored those comparable with control. In brief, our results showed that PAS-Na may have protective effects on learning and memory against Mn via increasing the number of ChAT-positive neurons and activity of ChAT protein.

Frequencies of D8S384 alleles and genotypes in European, African-American, Chinese, and Japanese populations.

D8S384 is a tetranucleotide tandem repeat locus. In order to evaluate the forensic validation of D8S384, the genotype distributions and allele frequencies in ten populations from three main ethnic groups were investigated, including Germans, Slovakians, African Americans, Japanese, and Chinese (Jilin, Guangzhou, Nanning, Hailaer, Dali, and Chengdu). A total of 1011 unrelated individuals, 41 pedigrees, 30 disputed paternity trios and three personal identification cases were analyzed for D8S384 by Amp-FLP technique. Many kinds of tissues, body fluids, secreta and stains have been tested. The alleles were determined by comparison with a human allele ladder. The results showed that D8S384 typing was both precise and reliable. There were eight alleles in these populations. The genotype distributions conformed to Hardy-Weinberg equilibrium predictions. No mutation events were observed. With a maximum likelihood method, the mutation rate was indirectly estimated as 2.14 x 10(-5). The heterozygosity was 0.704 +/- 0.014 at D8S384 locus. All these results suggest that D8S384 locus is a useful marker for forensic identification and paternity analysis.