Toxicity and Mechanism of Polygoni Multiflori Radix Alcohol Extract on L02 Cells / 中国实验方剂学杂志

Objective::To study the toxic effect of Polygoni Multiflori Radix alcohol extract (PME) on L02 cells and the mechanism of ROS inducing apoptosis via mitochondria pathway, so as to provide a basis for the rational and safe administration of Polygoni Multiflori Radix in clinic. Method::The 4, 5-dimethly-2-thiazolyl-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to detect the cell viability of PME at different concentrations (5, 10, 20 g·L-1). Nuclear morphology was observed by Hoechst 33342 staining. The apoptosis rate of cells was detected by Annexin V-FITC/PI. The release rate of lactate dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in the cells were detected by kit instruction. The changes of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were detected by flow cytometry. The relative protein expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), cysteinyl aspartate proteinase-9(proCaspase-9) and cysteinyl aspartate proteinase-3 (proCaspase-3) in the PME-administered group were detected by Western blot. Result::After treatment with PME at the concentration of 5, 10, 20 g·L-1, the survival rate of L02 cells were decreased in a concentration and time-depended manner. After treatment with PME for L02 cells, nucleus shrinkage, fragmentation and chromatin condensation were observed under fluorescence after Hoechst 33342 staining. Annexin V-FITC/PI double staining showed a upward cell apoptosis rate in PME 20 g·L-1 group. Compared with the normal control group, the release rate of LDH was significantly increased (P<0.01), the intracellular ROS level was significantly increased (P<0.01), and the SOD activity was significantly decreased (P<0.01), while the MMP rate was significantly decreased in PME 5, 10, 20 g·L-1 groups (P<0.05). With the increase in the concentration of PME, proCaspase-3, proCaspase-9, Bcl-2 protein showed a significantly downward trend in PME 10, 20 g·L-1 groups (P<0.01), while the expression of Bax protein was significantly up-regulated in PME 20 g·L-1 group (P<0.05). Conclusion::The study illustrated that PME have toxic effects on L02 cells, which may destroy the structure of hepatocytes to a certain extent, promote ROS levels, induce oxidative stress, activate the mitochondrial pathway, and then activate apoptosis-related proteins to cause cells damage. It is suggested that ROS-mediated mitochondrial pathway was involved in PME-induced apoptosis.

Acceptability of physical examination by male doctors in medical care: Taking breast palpation as an example / 华中科技大学学报（医学）（英德文版）

In this study, we conducted an investigation among medical workers, patients and college students concerning their acceptability of breast palpation performed by male doctors (hereinafter referred to as "acceptability", or "the examination", respectively, if not otherwise indicated), to get the information about their acceptability and reasons for accepting or declining the examination among the three population. A questionnaire investigation was conducted in 500 patients with breast diseases, 700 students of medical colleges, and 280 medical workers working in hospitals. The subjects were asked to choose between two options: accept or do not accept (the examination). The subjects were asked to fill out the questionnaire forms on free and anonymous basis and the questionnaire forms were collected on spot, immediately after completion. The questionnaires collected were coded, sorted out and checked. Data of the eligible questionnaires were input into Epidata software and analyzed by SPSS. Upon the establishment of the database, the intra-group data were tested by utilizing χ(2) test. Among 1480 questionnaires, 1293 (90.41%) questionnaires were retrieved. Our results showed that 56.78% of patients reported that they could accept breast palpation by male doctors. About 59.66% of medical staff expressed their acceptance of the examination, but only 35.03% of students said the examination. On the basis of this study, we were led to conclude that the examination is not well accepted by different populations, and therefore, (1) medical professionals and administrators should pay attention to the gender-related ethics in their practice and the feeling of patients should be respected when medical examinations involve private or sensitive body parts; (2) to this end, related departments should be properly staffed with doctors of both sexes, and this is especially true of the departments involving the examination or treatment of private or sensitive body parts; (3) health education should, among other things, include helping female patients to overcome the fear and anxiety in such examinations. This is of great importance since some women may miss the opportunity to get timely diagnosis.

Comparison of the mechanisms of intralesional steroid, interferon or verapamil injection in the treatment of proliferative scars / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of intralesional steroid, interferon alpha-2b or verapamil injection on proliferation, apoptosis and TGF-beta1 expression in keloid and hypertrophic scar in vivo.</p><p><b>METHODS</b>6 patients with keloids and 6 patients with hypertrophic scar were treated with intralesional injection of triamcinolone acetonide (40 mg/ml) or IFN alpha-2b (15 x 10(5) U/ml) or verapamil (2.5 mg/ml). Samples were collected on the 7th day after intralesional injection. Samples of untreated keloid and hypertrophic scar and normal skin were used as control. Expression of PCNA and TGF-beta1 was detected in situ by immunohistochemical staining, and apoptosis was detected in situ by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL).</p><p><b>RESULTS</b>1) Triamcinolone acetonide could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, as well as inducing apoptosis. 2) IFN alpha-2b could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, but not inducing apoptosis; 3) Verapamil could also prohibit proliferative scars through inhibiting proliferation and TGF-beta1 expression in fibroblasts, as well as inducing apoptosis. While the effect of inducing apoptosis was stronger than that of triamcinolone acetonide, the effect of inhibiting TGF-beta1 expression was weaker than those of triamcinolone acetonide and IFN alpha-2b.</p><p><b>CONCLUSIONS</b>Although intraleional injection of steroid, interferon alpha-2b or verapamil were all effective in the treatment of keloid and hypertrophic scar, their mechanisms are not similar.</p>

The role of cell pathway in apoptosis of fibroblasts from proliferative scar induced by steroid or IFN / 中华整形外科杂志

<p><b>OBJECTIVE</b>This paper is to investigate the effects of steroid or IFN alpha-2b on apoptosis and cell pathway of fibroblasts from keloids, hypertrophic scars and normal skins and different responses of different fibroblasts.</p><p><b>METHODS</b>6 samples from keloid, hypertrophic scar and normal skin were collected respectively and fibroblasts from different sources were cultured in vitro. After different fibroblasts were treated with dexamethasone (0.1 mg/ml) or IFN alpha-2b (1000 U/ml), Bax and Bcl-2 protein expressions were detected in situ by immunohistochemical staining; DNA ladders of different fibroblasts were observed by gel electrophoresis; and relative activated (phospho-) ERK1/2 and JNK pathways were detected by method of FACE ELISA.</p><p><b>RESULTS</b>Dexamethasone could induce apoptosis of fibroblasts from keloids, hypertrophic scars and normal skins through activating (phospho-) ERK1/2 and JNK pathways; IFN alpha-2b could not induce apoptosis of fibroblasts from different sources. IFN alpha-2b could inhibit (phospho-) ERK1/2 pathway and could not affect (phospho-) JNK pathways of fibroblasts from keloid and hypertrophic scar. IFN alpha-2b could affect neither (phospho-) ERK1/2 pathway nor (phospho-) JNK pathways of fibroblasts from normal skin.</p><p><b>CONCLUSIONS</b>The responses of different fibroblasts to steroid or IFN alpha-2b were different.</p>

Expression of nestin and neurogenin 3 in the human fetal pancreas / 中华外科杂志

<p><b>OBJECTIVE</b>To examine the expression of nestin and neurogenin 3 (Ngn3), the markers of pancreatic stem cells, in the human fetal pancreas.</p><p><b>METHODS</b>The human fetal pancreas tissue of 12 and 14 weeks were examined for the expression of nestin and Ngn3 using the techniques of immunofluorescence dye and RT-PCR.</p><p><b>RESULTS</b>Both nestin and Ngn3 expressed widely in 12 and 14 weeks before in human fetal pancreatic tissue. In these positive cells there was no co-expressing insulin or glucagon. There were nestin and Ngn3 co-expressing cells in ducts but not in the islets. The results of RT-PCR also indicated the expression of nestin and Ngn3.</p><p><b>CONCLUSIONS</b>There was no expression of the markers of mature endocrine cells in the nestin and Ngn3 positive cells, and they were the marks of no-differentiation cells in the human fetal pancreatic tissue.</p>