Objective To investigate the protectve effects and underlying mechanisms of deferroxamine on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal rat were used in a model of glutamate induced neurotoxicity.There were two experimental groups.Neurons were pretreated with deferroxamine before glutamate in the deferroxamine group, and neurons were treated with glutamate only in the control group.The morphological change was examined under microscope.Hoechst 33342 DNA staining method was used to study the ratio of condensed nuclei.The levels of lactate dehydrogenase (LDH), malonaldehyde (MDA) and hydroxyl radical were determined using biochemistry.The change in calcium signal was detected using microfluorescent technique.Results The neurons pretreated by deferroxamine had intact morphology with the ratio of condensed nuclei at 14% ± 6% compared to 58% ± 6% (t= 8.98, P ＜0.01 ) in the control group.LDH level was (36.42 ± 8.99) U/L in the deferroxamine group and was (68.06 ± 11.26) U/L in the control group ( t =3.25,P＜0.05).The respective levels of hydroxyl radical were (34.21 ±4.23) U/L and (47.06 ±8.79) U/L (t = 3.11, P ＜0.05 ).The respective levels of MDA were (12.26 ± 2.78 ) nmol/mg and (28.86±5.19) nmol/mg(t =4.88,P＜0.01).Conclusion Deferroxamine can protect neurons from glutamate induced damage.The mechanisms include an inhibition of Ca2+ overload and reduction in the levels of MDA and hydroxyl radicals.

Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly increased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.

Objective To investigate the role of Notch signaling in the progression of peritoneal fibrosis in a rat model induced by high glucose dialysate. Methods Male Sprague Dawley rats were subjected to daily peritoneal dialysis (PD) with a lactate-buffered solution containing 4.25% glucose. They were sacrificed at 2 and 4 weeks after PD. The parietal thickness was measured with Masson staining. The expression of TGF-β1, E-cadherin, α-SMA and collagen Ⅰ was examined by immunoblotting. The expression of Notch ligand Jagged-1 and the negative Notch signaling regulato--Numb was analyzed by both immunoblotting and RT-PCR. The expression of a Notch nuclear target gene Hcs-1 was examined by RT-PCR. Results Both HE and Masson trichrome staining revealed an increase in peritoneal thickness with a loss of mesothelial cells and a rich of collagen matrix deposition in the submesothelial zone was evident at 4 weeks after PD. Meanwhile, compared to healthy rats, the expression of TGF-β1, ct-SMA and collagen Ⅰ was significantly increased, but the expression of E-cadherin was decreased in peritoneum after PD treatment. It was difficult to detect the Jagged-1 and Hes-1 expression in normal peritoneum, but their expression was graduaUy increased after PD. In contrast, the expression level of Numb, a negative regulator of Notch signaling, was dramatically decreased after PD. Conclusions Notch signaling is activated during the process of PD-induced peritoneal fibrosis and the activation of Notch signaling is associated with the loss of negative regulation of Notch signaling via decreased expression of Numb. Inhibition of Notch signaling via overexpression of its negative regulators such as Numb may be a novel therapeutic approach for peritoneal fibrosis in PD patients.

Objective Tumor suppressor gene p53 can inhibit tumor cell growth, arrest cell cycle, and promote apoptosis.Howev-er, the effects of p53 on the pathogenesis of breast cancer have not been fully elucidated.The aim of this study was to explore the expression of p53 protein and the correlation with clinical pathologic features in breast cancer.Furthermore, the regulatory effects of 5-aza-2′-deoxycyti-dine on p53 in breast cancer cell line were also studied. Methods The expression of p53 protein in 80 cases of breast cancer and normal and adjacent tissue were determined by the immunohistochemical staining .The expressions of p53 mRNA and p53 protein in breast cancer cell line were determined by RT-PCR and Western blotting. Results The positive rate of p53 in breast cancer (41.25%) was higher than that in the normal and adjacent tissue (22.5%) (P<0.01).The expression of p53 was not significantly correlated with age, grade, stage and lymph node metastasis (P>0.05).The low expression of p53 both in mRNA and in protein levels were found in breast cancer cell line of MCF-7.The expres-sion of p53 increased after 5-aza-2′-deoxycytidine administration . Conclusion p53 is highly expressed in breast cancer , which may play an im-portant role in the development and progression of breast cancer. 5-aza-2′-deoxycytidine, up-regulating the p53 expression in breast cancer cell line, which provides the evidents for the development of therapeutic drugs for the patients with low expression of p53 breast cancer.

Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly increased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.
Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Osteoblasts ; drug effects ; metabolism ; Osteogenesis ; drug effects ; RANK Ligand ; metabolism ; Rats ; Signal Transduction ; drug effects ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

BACKGROUNDThe Borg scale is most commonly used to measure dyspnea in China. However, many patients that find it is difficult to distinguish the labeled numbers corresponding to different dyspnea scores. We developed a new method to rate dyspnea, which we call the count scale (CS). It includes the count scale number (CSN) and count scale time (CST). The aims of the present study were to determine the reproducibility and sensitivity of the CS during exercise in patients with chronic obstructive pulmonary disease (COPD).

METHODSFourteen male patients with COPD (aged 58.00 ± 7.72 years) participated in this study. A progressive incremental exercise and a 6-minute constant work exercise test were performed every 2 to 3 days for a total of 3 times. The CS results were evaluated at rest and at 30% and 70% of maximal workload (Wmax) and Wmax. The Borg scales were obtained during exercise.

RESULTSNo significant differences occurred across the three trials during exercise for the CS and Borg scores. The CSN and CST were more varied at Wmax (coefficient of variation (CV) = (22.28 ± 16.96)% for CSN, CV = (23.08 ± 19.11)% for CST) compared to 30% of Wmax (CV = (11.92 ± 8.78)% for CSN, CV = (11.16 ± 9.96)% for CST) and 70% of Wmax (CV = (9.08 ± 7.09)% for CSN, CV = (12.19 ± 12.32)% for CST). Dyspnea ratings with either CSN or CST tended to decrease at the higher workload compared to the lower workload. CSN and CST scores were highly correlated (r = 0.861, P < 0.001). CSN was negatively correlated with Borg scores (r = -0.363, P = 0.001). Similar results were obtained for the relationship between CST and Borg scores (r = -0.345, P = 0.003).

CONCLUSIONWe concluded that the CS is simple and reproducible when measuring dyspnea during exercise in patients with COPD.

Aged ; Dyspnea ; diagnosis ; Exercise ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; Reproducibility of Results

Objective To analyze the correlation between femoral head CT values in routine abdominopelvic CT scanning and hip bone mineral density(BMD)values in hip quantitative CT(QCT)measurements,and to investigate the diagnostic value of femoral head CT values for hip osteoporosis.Methods Totally 176 patients undergoing non-enhanced abdominopelvic CT scanning at some hospital from June 1,2021 to January 20,2022 were selected prospectively,and femoral head CT values and hip BMD values were measured on the scanned images.The patients were divided into a normal bone mass group,a reduced bone mass group and an osteoporosis group according to hip T values and the"China Guideline for the Diagnosis Criteria of Osteoporosis with Quantitative Computed Tomography(2018)".Analyses were carried out for the correlation between femoral head CT values and hip BMD values,the difference between femoral head CT values and BMD values of the two sides with paired t-test,the difference of CT values between the three groups with ANOVA and the diagnostic efficacy of femoral head CT values for diagnosing hip osteoporosis with AUC curve.Results The values of femoral head CT and hip BMD had a strong correlation(r=0.696),which were slightly higher on the left than on the right.The three groups had statistically significant differences of femoral head CT values at two sides(P=0.000),which were ranked according to CT values as the normal bone mass group,the reduced bone mass group and the osteoporosis group.Femoral head CT values and hip BMD values both reached the peak at the age of 41 to 50 years,and then decreased gradually.Femoral head CT values behaved well in predicting hip osteoporosis,with femoral head AUC value being 0.859 on the left and 0.846 on the right.Conclusion The values of femoral head CT and hip BMD had a strong correlation,and femoral head CT values are valuable for diagnosing osteoporosis.[Chinese Medical Equipment Journal,2023,44(9):64-68]

BACKGROUNDIt is widely recognized that the diagnosis of parathyroid carcinoma (PC) is often difficult because of the overlap of characteristics between malignant and benign parathyroid tumors, especially at an early stage. Our study aimed to investigate the differential expression of Ki-67, galectin-3, fragile histidine triad (FHIT) gene, and parafibromin in PC, parathyroid adenoma (PA), parathyroid hyperplasia (PH), and normal parathyroid (NP) tissues; then to assess these expression values for use in differential diagnosis of malignant and benign parathyroid tumors.

METHODSData of 15 cases with PC, 19 PAs, and 8 PHs were retrospectively analyzed for their clinical characteristics. The expression of Ki-67, galectin-3, FHIT, and parafibromin were detected via immunohistochemistry in the above-mentioned specimens and 6 NPs as control.

RESULTSComplete loss of parafibromin expression was seen in 9 of 15 (60%) carcinomas, and all normal parathyroid tissues and parathyroid benign tumors stained positive for parafibromin except for one (4%) adenoma. Galectin-3 staining was positive in 11 of 15 (73%) carcinomas, 5 of 19 (26%) adenomas, 1 of 8 (12%) hyperplasias, and 0 of 6 normal tissues. The Ki-67 proliferative index was high in 4 of 15 (27%) carcinomas, 1 of 19 (5%) adenomas, and none of the hyperplasia or normal tissues. FHIT expression did not differ appreciably among the tumor types. The combination of overexpression of galectin-3 or loss of parafibromin increased sensitivity for PC to 87%, while the specificity of both positive galectin-3 and positive Ki-67 could reach 100%.

CONCLUSIONSThese data suggested that loss of parafibromin and overexpression of galectin-3 and Ki-67 might help to distinguish parathyroid carcinoma from other parathyroid tumors. And the combination of two or three of these markers might produce better sensitivity and/or specificity for the diagnosis of parathyroid carcinoma.

Acid Anhydride Hydrolases ; metabolism ; Galectin 3 ; metabolism ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Neoplasm Proteins ; metabolism ; Parathyroid Neoplasms ; metabolism ; Tumor Suppressor Proteins ; metabolism

The 2017 China (Lianyungang) International Medical Technology Conference was held in Lianyungang,Jiangsu Province during November 15-17,2017.During this conference,the Division for Traditional Chinese Medicine and Natural Products Pharmacology of Chinese Pharmacological Society (CNPHARS) and Jiangsu Kanion Pharmaceutical Co. Ltd.jointly held the Forum on R&D and Interna-tionalization of New Drugs and Health Products of Traditional Chinese Medicine.The forum was co-chaired by Professor ZHANG Yong-xiang, President of CNPHARS, Chair of Division for Traditional Chinese Medicine and Natural Products Pharmacology of CNPHARS,and Chair of the Natural Product Section of Inter-national Union of Basic&Clinical Pharmacology(IUPHAR), Professor DU Guan-hua,former President of CNPHARS and Vice-Chair of Division for Traditional Chinese Medicine and Natural Products Pharmacology of CNPHARS,and Dr.XIAO Wei,Chairman of the Board of Jiangsu Kanion Pharmaceutical Co. Ltd. And Vice-Chair of Division for Traditional Chinese Medicine and Natural Products Pharmacology of CNPHARS. More than 70 scholars attended the forum, including four foreign experts [Michael SPEDDING, Secretary-General of IUPHAR; Professor Valérie B. SCHINI-KERTH, Vice-Chair of the Natural Product Section of IUPHAR; Professor Cherry WAINWRGHT, Director of Centre for Natural Product Drugs of Robert Gordon University; Professor InKyeom KIM, Director of the Korean Society of Pharmacology], members of the Division for Traditional Chinese Medicine and Natural Products Pharmacology of CNPHARS and leading researchers at Jiangsu Kanion Pharmaceutical Co.,Ltd.GU Jin-hui,Director of the Division of National Science and Technology Major Project for Drug Innovation,Department of Health Science,Technology and Education,National Health and Family Planning Commission of the People's Republic of China was also invited to attend the forum. Representatives discussed the R&D and internationalization of new drugs and health products of traditional Chinese medicine.The summary of views and advice of some experts was published here for the purpose of promoting domestic and overseas academic exchange, and playing an active role in improving the level of R&D and internationalization of new drugs and health products of traditional Chinese medicine in China.

As an important part of epigenetic marker, DNA methylation involves in the gene regulation and attracts a wide spread attention in biological auxology, geratology and oncology fields. In forensic science, because of the relative stable, heritable, abundant, and age-related characteristics, DNA methylation is considered to be a useful complement to the classic genetic markers for age-prediction, tissue-identification, and monozygotic twins' discrimination. Various methods for DNA methylation detection have been validated based on methylation sensitive restriction endonuclease, bisulfite modification and methylation-CpG binding protein. In recent years, it is reported that the third generation sequencing method can be used to detect DNA methylation. This paper aims to make a review on the detection method of DNA methylation and its applications in forensic science.
CpG Islands ; DNA Methylation/genetics* ; Epigenesis, Genetic ; Epigenomics ; Forensic Genetics/trends* ; Genetic Markers/genetics* ; Humans ; Sulfites ; Twins, Monozygotic/genetics*