Regulation of Wenjingtang on HIF-1α Expression and Mitochondrial Function of Ectopic Endometrial Stromal Cells in Patients with Endometriosis / 中国实验方剂学杂志

ObjectiveThis study investigated the mechanism of Wenjingtang in the prevention and treatment of endometriosis （EMT） from the perspective of regulating hypoxia stress and mitochondrial function. MethodPrimary human endometrial stromal cells （ESCs） form ectopic endometrial tissues were isolated and cultured， the cells were divided into control group （Control）， 5% control serum group （5% KBXQ）， 10% control serum group （10% KBXQ）， 5% Wenjingtang serum group （5% WJTXQ） and 10% Wenjingtang serum group （10% WJTXQ）. ESCs in different groups were detected for proliferation by methyl thiazolyl tetrazolium （MTT） assay， mRNA and protein expression of hypoxia inducible factor-1α （HIF-1α） by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot analysis， mitochondrial ultrastructure by transmission electron microscope， mitochondrial function ［mitochondrial membrane potential， mitochondrial membrane potential（MMP） and cytochrome C（Cyt C） content］ and apoptosis （cell membrane permeability， nuclear fluorescence intensity， nuclear size and cell counts） by high content screening （HCS） assay， apoptosis rate by flow cytometry， and proteins of B-cell lymphoma-2 （Bcl-2） associated X （Bax）， Bcl-2 and cleaved cysteinyl aspartate specific proteinase-3 （cleaved Caspase-3） by Western blot. ResultCompared with Control group， the 5% KBXQ and 10% KBXQ groups showed increased cell viability （P<0.01）， there was no significant change in HIF-1α mRNA and protein expression， transmission electron microscopy showed that the mitochondrial cristae were obvious and the inner and outer membranes of mitochondria were clear， HCS multichannel fluorescence staining showed that there were no significant changes in the expression of MMP， Cyt C and cell membrane permeability， and the nuclei showed uniform light staining， there were no significant changes in apoptosis rate， cleaved Caspase-3 protein expression and Bax/Bcl-2 ratio. Compared with Control group and corresponding concentration KBXQ group， the 5% WJTXQ and 10% WJTXQ group showed decreased cell viability （P<0.01） and HIF-1α mRNA and protein levels （P<0.05，P<0.01）， the ultrastructure of mitochondria was destroyed， some mitochondria were swollen and the cristae were blurred， moreover， decreased MMP and up-regulated Cyt C release （P<0.05，P<0.01）， increased cell membrane permeability （P<0.01）， and apoptosis characteristics included nuclear pyknosis， DNA agglutination in nucleus and decrease of cell numbers were observed （P<0.05，P<0.01）， increased apoptosis rate （P<0.01）， which was consistent with the results of HCS analysis， and up-regulated expression levels of cleaved Caspase-3 protein and Bax/Bcl-2 ratio （P<0.05，P<0.01）. ConclusionIn conclusion， the results suggest that Wenjingtang can improve hypoxia stress via down-regulating HIF-1α expression in ectopic ESCs， and inhibit cell proliferation， reduce mitochondrial biological activity and induce apoptosis， which might be the internal mechanism of Wenjingtang in preventing and treating EMT.

Effects of TNF-α on ICAM-1 and LFA-1 expression in peripheral blood mononuclear cells of children with febrile seizures / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of TNF-α on ICAM-1 and LFA-1 expression in peripheral blood mononuclear cells (PBMC) of children with febrile seizures (FS).</p><p><b>METHODS</b>Sixteen children with FS and 16 age- and gender-matched healthy children were enrolled. The samples of PBMC from FS children were randomized into two groups with or without TNF-α treatment (TNF-α concentration 1.0 ng/mL). PBMC were purified and cultured with a conventional method in vitro. The expression of ICAM-1 and LFA-1 in PBMC was determined by flow cytometry (FCM).</p><p><b>RESULTS</b>ICAM-1［(20±9)% vs (14±7)%)］and LFA-1［(43±16)% vs (30±16)%］expression in PBMC in the untreated FS group was significantly higher than that in the normal control group (P<0.05). Compared with the untreated FS group, the treatment with TNF-α remarkably increased the ICAM-1 expression［(27±11)%］(P<0.05). PBMC LFA-1 expression［(52±21)%］in the TNF-α-treated group was higher than that in the untreated FS group, although there were no statistical differences between the two groups.</p><p><b>CONCLUSIONS</b>TNF-α treatment may increase LFA-1 and ICAM-1 expression in PBMC of children with FS.</p>

Anxiety and its effect factors in children with epilepsy / 中华神经医学杂志

Objective To investigate the anxiety state of children with epilepsy and explore the influence of patients' depression, personality and guardians' anxiety, depression and personality on the patients' anxiety state. Methods Ninety-five epileptic children (epilepsy group) and their guardians were assessed to rate their anxiety, depression and personality with Social Anxiety Scale for Children (SASC), Depression Self Rating Scale for Children (DSRS), Self-Rating Anxiety Scale (SAS), Beck Depression Inventory (BDI) and Eysenck Personality Questionnaire of children and adult (EPQ). One hundred and eighteen children recovered from upper respiratory infection were included in our study as control group. Results The SASC scores in the epilepsy group were higher than those in the control group (P＜0.05); in the epilepsy group, the SASC scores of girls were higher than those of boys (P＜0.05);the anxiety level of patients of all ages increased with ages; SASC scores in epilepsy group aged 13-15 years were higher than that of control group (P＜0.05). The SASC scores were positively correlated to the DSRS scores and the scores of N and P dimension of EPQ, and negatively correlated to scores of E dimension of EPQ in the epileptic group (P＜0.05). The SASC scores of patients were positively correlated to the SAS scores of guardians and negatively to scores of E dimension of EPQ in guardians (P＜0.05). No correlation between SASC scores of patients and scores of guardians' BDI and P, N dimensions of EPQ was noted (P＞0.05). Under the condition that educational level of guardians were the same, no significant differences in the SASC scores were noted between epilepsy group and control group (P＞0.05). Under the condition that educational level of guardians were different, no significant differences in the SASC scores were noted in the epilepsy group (P＞0.05). Under the condition that the occupation of guardians were the same, no significant differences in the SASC scores were noted between epilepsy group and control group (P＞0.05). Although the guardians' occupation was different (workers, farmers or administration staff), no significant differences in their children's SASC scores were noted in the epilepsy group (P＞0.05). The epileptic seizure type (partial or generalized seizure), neuroimaging changes and course of the disease had no significant influence on the patients' SASC scores in the epilepsy group (P＞0.05). Conclusion Anxiety level of children with epilepsy is higher than that of the healthy ones, which is related to gender,and age of children, anxiety or depression level and some personality of themselves and guardians, but not correlated to epileptic seizure type, neuroimaging changes, course of the disease, guardians' educational level and occupation.