Effects of TGF-ß1 on the expression of endometrial stromal cell-related protein and mRNA.

OBJECTIVE: To investigate the effects of transforming growth factor ß1 (TGF-ß1) on α-smooth muscle actin (α-SMA), insulin-like growth factor I (IGF-I), and type I collagen (Col I) expression in endometrial stromal cells as well as on fibronectin (FN) level. PATIENTS AND METHODS: 56 patients with normal endometrial tissue obtained from surgery were selected from June 2018 to November 2019. Endometrial stromal cells were isolated from patients and then assigned to the control group and observation group (addition of TGF-ß1) followed by the analysis of cellular activity by Thiazole blue staining; and α-SMA, IGF-I, Col I, and FN mRNA and protein levels by real-time fluorescent PCR and Western blot. RESULTS: The cell proliferation rate at 12 h, 24 h, 36 h, and 72 h after culture in both groups was higher than 0 h (p < 0.05) with higher cell proliferation in the observation group than the control group (p < 0.05). Real-time fluorescence PCR results showed that the levels of α-SMA, IGF-I, Col I, and FN mRNA in endometrial stromal cells of the observation group after TGF-ß1 intervention were higher than those in the control group (p < 0.05). Meanwhile, α-SMA, IGF-I, Col I, and FN protein level was also elevated in the observation group after TGF-ß1 treatment (p < 0.05). CONCLUSIONS: TGF-ß1 can stimulate the proliferation of endometrial stromal cells, which may be related to regulate α-SMA, IGF-I, Col I, and FN expression.

Association between hsa_circ_0006156 expression and incidence of gastric cancer.

OBJECTIVE: To screen the differentially expressed circular ribonucleic acids (circRNAs) related to gastric cancer and to explore their associations with the clinicopathological features of gastric cancer. PATIENTS AND METHODS: Cancer tissues of 50 gastric cancer patients undergoing surgical resection in our hospital from April 2015 to December 2018 were collected as an experimental group, while the para-carcinoma tissues were used as the control group. First, the differentially expressed circRNAs were screened by analyzing the circRNA profile in the microarray. Then, the expression of hsa_circ_0006156 in tissues was detected via Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) in both groups. The potential associations of the relative expression level of hsa_circ_0006156 with clinicopathological features and prognosis were analyzed according to the clinical data of gastric cancer patients. RESULTS: Six significantly downregulated circRNAs in gastric cancer patients were screened out. The results of RT-qPCR showed that the expression level of hsa_circ_0006156 was significantly lower in gastric cancer tissues than that in para-carcinoma tissues (p<0.05). Accordingly, 50 gastric cancer patients were divided into hsa_circ_0006156 high expression group and hsa_circ_0006156 low expression group based on the fold change of hsa_circ_0006156 in para-carcinoma tissues than that of gastric cancer tissues (fold change>3). The expression level of hsa_circ_0006156 was not correlated with the age and gender of gastric cancer patients (p>0.05) but correlated with the lymph node metastasis (p<0.05), nerve invasion (p<0.05), and degree of tumor differentiation (p<0.05). The expression level of hsa_circ_0006156 was also significantly associated with the progression-free survival (PFS) and overall survival (OS) of patients (p<0.05). According to the multivariate analysis of variance, the PFS of gastric cancer patients was associated with nerve invasion, lymph node metastasis, and hsa_circ_0006156 expression (relative risk coefficient=1.742, 2.329, and 3.003). Meanwhile, the OS was associated with lymph node metastasis, nerve invasion, degree of tumor differentiation, and hsa_circ_0006156 expression (relative risk coefficient =1.604, 2.405, 2.114, and 2.004). Moreover, the survival analysis revealed that PFS was markedly prolonged in the hsa_circ_0006156 high expression group compared with that in the hsa_circ_0006156 low expression group. CONCLUSIONS: The expression of hsa_circ_0006156 substantially declines in gastric cancer tissues, which is related to the differentiation degree, presence, or absence of lymph node metastasis and prognosis of gastric cancer patients. Therefore, hsa_circ_0006156 may clinically serve as a biomarker for the prognostic prediction of gastric cancer patients.

Effects of parathyroid hormone-related protein on osteogenic and adipogenic differentiation of human mesenchymal stem cells.

OBJECTIVE: This work aims to investigate the effects of parathyroid hormone-related peptide (PTHrP) (1-86) on osteogenic and adipogenic differentiation of human mesenchymal stem cells (hMSCs) and the related mechanisms. MATERIALS AND METHODS: hMSCs were isolated and cultured in vitro. They were divided into control group, osteogenesis group, adipogenesis group, osteogenesis+PTHrP group and adipogenesis+PTHrP group. The cell proliferation and differentiation, and expression levels of osteopontin (OPN) and lipoprotein lipase (LPL) mRNA were observed. RESULTS: The proliferation rates of hMSCs in osteogenesis+PTHrP and adipogenesis+PTHrP group were significantly higher than that in control group, respectively (p < 0.01). The alkaline phosphatase (ALP)-positive osteoblasts firstly appeared in osteogenesis+PTHrP group, and Sudan IV-positive adipocytes firstly appeared in adipogenesis group. The expression level of OPN mRNA in osteogenesis+PTHrP group was significantly higher than that in osteogenesis group (p < 0.05), and that in adipogenesis+PTHrP group was also higher than adipogenesis group (p < 0.05). The expression level of LPL mRNA in osteogenesis+PTHrP group was significantly lower than that in osteogenesis group, and that in adipogenesis+PTHrP group was also lower than adipogenesis group (p < 0.05). CONCLUSIONS: The osteogenesis and adipogenesis are related to each other during the induced differentiation of hMSCs. PTHrP (1-86) can promote the osteogenic differentiation and inhibits the adipogenic differentiation for hMSCs.

Toxicity of a novel anti-tumor agent 20(S)-ginsenoside Rg3: a 26-week intramuscular repeated administration study in rats.

The purpose of this study is to investigate the potential subchronic toxicity of 20(S)-Ginsenoside Rg3(Rg3), by a 26-week repeated intramuscular administration in rats. Rg3 was administrated to rats at dose levels of 0, 4.2, 10.0 or 20.0 mg/kg/day. There was no treatment-related mortality and, at the scheduled autopsy, dose-dependent increases in the absolute and relative spleen weights, of both the 10.0 mg/kg and 20.0 mg/kg dose groups were observed. Absolute and relative kidney weights were significantly elevated in the female 10.0 mg/kg dose group and in the male 20.0 mg/kg dose group. Hematological investigations revealed a dose-dependent increase in the total white blood cell (WBC) count and in the percentage of neutrophils, but a decrease in the percentage of lymphocytes, in rats treated with doses of 10.0/20.0 mg/kg. These effects were completely reversible during the recovery period, and no other adverse effects were observed. It was concluded that the 26-week repeated intramuscular dose of Rg3 caused increases in the spleen and kidney weights, WBC counts and in the percentage of neutrophils, but a decrease in the percentage of lymphocytes, with doses of 10.0 or 20.0 mg/kg/day. The no-observed-adverse-effect level for rats was considered to be 4.2 mg/kg/day.

Determination of glycyrrhetic acid in human plasma by HPLC-MS method and investigation of its pharmacokinetics.

OBJECTIVE: To develop a high performance liquid chromatography mass spectrometry (HPLC-MS) method for the determination of the glycyrrhetic acid (GA) in human plasma and for the investigation of its pharmacokinetics after the oral administration of 150 mg diammonium glycyrrhizinate test and reference capsule formulations. METHODS: The GA in plasma was extracted with ethyl acetate, separated on a C(18) column with a mobile phase of methanol (5 mmol/L ammonium acetate)-water (85 : 15, V/V) and analysed using a MS detector. Ursolic acid (UA) was used as internal standard. The target ions were m/z 469.5 for GA and m/z 455.6 for UA, the fragment voltages were 200 V and 100 V for GA and UA respectively. RESULTS: The calibration curve was linear over the range of 0.5-200 ng/mL (r = 0.9974). The limit of quantification for GA in plasma was 0.5 ng/mL, the recovery was 76.0-80.0%, and the inter- and intra-day relative standard deviations (RSD) were <12%. The pharmacokinetic parameters of GA after a single dose of 150 mg diammonium glycyrrhizinate test and reference were as follows: the half life (t(1/2)) 9.65 +/- 3.54 h and 9.46 +/- 2.85 h, the time to peak concentration (T(max)) 10.95 +/- 1.32 h and 11.00 +/- 1.30 h, the peak concentration (C(max)) 95.57 +/- 43.06 ng/mL and 103.89 +/- 49.24 ng/mL; the area under time-concentration curve (AUC(0-48) and AUC(0-infinity)) 1281.84 +/- 527.11 ng.h/mL and 1367.74 +/- 563.27 ng.h/mL, 1314.32 +/- 566.40 ng.h/mL and 1396.97 +/- 630.06 ng.h/mL. The relative bioavailability of diammonium glycyrrhizinate capsule was 98.88 +/- 12.98%. CONCLUSION: The assay was sensitive, accurate and convenient, and can be used for the determination of GA in human plasma. Comparison of the bioavailability and pharmacokinetic profile of GA indicated that the test and reference capsules were bioequivalent.