miR-638 Serves as a Biomarker of 5-Fluorouracil Sensitivity to Neoadjuvant Chemotherapy in Breast Cancer / 한국유방암학회지

Purpose@#Neoadjuvant chemotherapy (NAC) is widely used to treat breast cancer (BC). The prediction and evaluation of chemotherapy responses remains a significant challenge. @*Methods@#MicroRNAs (miRNAs) play a crucial role in cancer drug resistance. We used a miRNA microarray and identified that miR-638 is downregulated in chemoresistant cases.However, the exact role of miR-638 and the underlying mechanisms of chemoresistance remain unclear. Using real-time quantitative reverse transcription polymerase chain reaction, we found significant downregulation of miR-638 in chemoresistant patients compared with chemosensitive patients. To explore the function of miR-638, we overexpressed and inhibited miR-638 expression in MDA-MB-231 and MCF-7 cells by transfecting them with miR-638 mimics and miR-638 inhibitor, respectively. Cell proliferation and apoptosis were measured using MTS and flow cytometry, respectively. A minimal patient-derived xenograft (MiniPDX™) model was established to evaluate the chemosensitivity to different drugs. @*Results@#The results showed that cell proliferation decreased and cell apoptosis increased in cells transfected with the miR-638 mimic, and cell proliferation and apoptosis were reversed with transfection of miR-638 inhibitor compared with the control group. Among patients who received 5-fluorouracil (5-FU), miR-638 expression levels were lower in the chemoresistant group than in the chemosensitive group. The MiniPDX™ model showed that MDA-MB-231 cells overexpressing miR-638 were more susceptible to 5-FU treatment in vivo. @*Conclusion@#We provided evidence of acquired resistance to 5-FU caused by miR-638 deficiency. Alterations in miR-638 may be used with 5-FU chemotherapy during NAC for BC.

Role of matrix metalloproteinase-9 down-regulation in learningand memory dysfunction induced by propofol treatment in rats / 中国药理学通报

Aim To investigate the role of matrix metalloproteinase-9 down-regulation in the learning and memory dysfunction induced by propofol treatment in rats.Methods 7-day-old SD rats were randomly divided into three groups(n=18):control group(NS group) and repeated doses of propofol group(RP group) was intraperitoneally injected with normal saline and propofol respectively for consecutive seven days, single dose of propofol group(SP group) were intraperitoneally injected with normal saline first for consecutive six days, and then injected with propofol on 7th day.The blood gas and glucose levels were monitored of six rats randomly selected from each group.Morris water maze was conducted to test the learning and memory functions of the remaining rats.The expression of MMP-9, BDNF and caspase-3 was detected by Western blot, and the hippocampal neuron apoptosis was determinated by TUNEL staining.Results Compared with NS group and SP group, the escape latency in RP group was prolonged significantly, exploration time and the number of crossing the platform in RP group were markedly decreased(P0.05).Conclusions Repeated exposure to propofol can lead to a decline in long-term learning and memory functions in neonatal rats, which may be related to the down-regulation of MMP-9 expression, proBDNF and mBDNF conversion disorder in hippocampus and the apoptosis of hippocampal neurons.However, single exposure to propofol has no significant effect.

Effects of propofol on rat brain function and tPA/MMP9 expression / 中国病理生理杂志

AIM: To investigate the effects of propofol on the expression of tissue-type plasminogen activator (tPA) and matrix metalloproteinase 9 (MMP9) in the hippocampus and the cognitive function in neonatal rats.METHODS: The 7-day-old rats were randomly divided into 3 groups: the rats in control (CON) group were intraperitoneally injected with normal saline for 7 d;the rats in single dose of propofol anesthesia (SP) group were intraperitoneally injected with normal saline for 6 d and with propofol on the 7th day;the rats in repeated dose of propofol anesthesia (RP) group were intraperitoneally injected with propofol for 7 d.Blood glucose and blood gas analysis were tested in 6 rats of each group.The rats were randomly selected from each group to isolate the hippocampal tissues at 2 h, 24 h, 48 h, 72 h and 30 d after the last injection.The spatial learning and memory functions of the other rats aged 25 d were determined by Morris water maze.The morphological changes of the hippocampus were observed by HE staining and Nissl's staining.The expression of tPA and MMP9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS: Compared with group CON, the protein expression of tPA and MMP9 in RP group was significantly decreased at each time point, while no significant decrease was observed in SP group except at the time point of 24 h.Compared with CON group, the mRNA expression of tPA and MMP9 was down-regulated obviously in RP group, which was not significantly down-regulated in SP group.From the 3rd training day of Morris water maze beginning, the escape latency was prolonged, and the space exploration time and the number of crossing the original platform location were reduced in RP group compared with CON group and SP group, while no significant difference was observed between CON group and SP group.Compared with CON group, the number of nerve cells reduced and nerve cells arranged in disorder in the hippocampus in RP group.Moreover, the number of Nissl body decreased significantly and finally developed into neuronal degeneration and necrosis in RP group, and no significant difference between SP group and CON group was observed.CONCLUSION: Repeated dose of propofol anesthesia leads to long-term cognitive dysfunction in neonatal rats, which may be related to the down-regulation of tPA and MMP9 expression and destruction of normal morphology and function of neurons in hippocampus, whereas single dose of propofol anesthesia has no such effects.

Effect of propofol anesthesia on the expression of BDNF mRNA and proBDNF/mBDNF in hippocampus and long-term spatial learning/memory functions in neonatal rats / 解放军医学杂志

Objective To investigate the effect of propofol anesthesia on the expression of brain-derived neurotrophic factor (BDNF) mRNA and BDNF precursor (proBDNF)/mature (mBDNF) in hippocampus of neonatal rats, and the relationship with long-term spatial learning/memory functions. Methods One hundred and eight SD rats aged 7 days weighing 12-16g were randomly divided into 3 groups (36 each): control group (C), propofol anesthesia once group (P1) and multiple propofol anesthesia group (P2). Rats in group C were intraperitoneally injected with 0.9% normal saline, 7.5ml/kg once a day for 7 days; rats group P1 were injected normal saline 7.5ml/kg once a day for 6 days and propofol 75mg/kg on the last day; rats group P2 received propofol 75mg/kg once a day for 7 days. Fifteen minutes after the last intraperitoneal injection, blood gas analysis and blood glucose detection were performed in 6 rats of each group. The hippocampal tissues were taken at 6, 24, 48, 72h and 30d to measure BDNF mRNA, proBDNF and mBDNF by RT-PCR and Western blotting after the completion of the injection. Learning and memory functions were assessed using Morris water maze at 25 days old of rats. Results The expression of BDNF mRNA and mBDNF protein increased gradually in hippocampus, while of proBDNF protein and the ratio of proBDNF/mBDNF decreased gradually in group C. In group P1, the BDNF mRNA and mBDNF protein were down-regulated, the ratio of proBDNF/mBDNF were up-regulated (P<0.05), the escape latency was prolonged, and space exploration time was shortened compared with group C (P<0.05). Compared with group C and group P1, the BDNF mRNA and mBDNF protein were down-regulated significantly, the proBDNF protein and the ratio of proBDNF/mBDNF were up-regulated observably (P<0.05), the escape latency was distinctly prolonged, and space exploration time was shortened apparently in group P2 (P<0.05). Conclusions Propofol anesthesia may impair the learning and memory function of infancy in rats, the effects are more obvious in repeated propofol anesthesia than in single ones. The mechanisms may be related to the changed expression of BDNF mRNA and the ratio of proBDNF/mBDNF.

Relationship of sperm morphology with the outcomes of in vitro fertilization and embryo transfer / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the application value of normal sperm morphology on the outcomes of classic in vitro fertilization and embryo transfer (IVF-ET).</p><p><b>METHODS</b>This study included 659 infertile couples admitted to our center for IVF-ET. Based on the percentage of morphologically normal sperm (MNS), we divided the patients into groups A (n = 112, MNS < 2%), B (n = 180, MNS > or = 2 - < 4%), C (n = 74, MNS > or = 4 - < 5%), and D (n = 293, MNS > or = 5%), and compared the rates of fertilization, normal fertilization, embryos obtained, biochemical pregnancy, clinical pregnancy, implantation, and live birth among different groups.</p><p><b>RESULTS</b>The mean fertilization rate was significantly higher in groups C (71.90%) and D (72.89%) than in A (57.97%) and B (63.29%) (P < 0.05), with no remarkable differences either between A and B (P > 0.05) or between C and D (P > 0.05). The normal fertilization rate was also significantly higher in group D (57.16%) than in A (46.52%) and B (50.89%) (both P < 0.05) as well as in C (54.67%) than in A (P < 0.05). The rate of embryos obtained, too, was markedly higher in group D (55.62%) than in B (45.75%) (P < 0.05), but none with remarkable difference from other groups (all P > 0.05). There were no statistically significant differences among the four groups in the rates of biochemical pregnancy, clinical pregnancy, implantation, abortion, and live birth (all P > 0.05).</p><p><b>CONCLUSION</b>The rate of MNS had some influence on IVF-ET, and 5% MNS exhibited a higher value than 4% MNS in predicting the outcomes of IVF.</p>

Expression of microRNA-100 in human gastric cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of microRNA-100 (miR-100) in human gastric cancer cells and its role of miR-100 in migration, invasion and proliferation in gastric cancer cell line SGC7901.</p><p><b>METHODS</b>Total RNAs were extracted from formalin-fixed and paraffin-embedded tissue samples of SGC7901 cells, gastric cancer (50 cases), non-tumor (18 cases) and lymph nodes with metastases (18 cases). The expression of miR-100 was examined by reverse transcription (RT)-qPCR. Additionally, SGC7901 cells were transfected with Pre-miR-100 and negative control constructs, and then their ability of migration, invasion and proliferation in vitro was documented after 48 hours.</p><p><b>RESULTS</b>RT-qPCR showed that although miR-100 expressed in all samples, compared to non-tumour tissues, the expression was lower both in SGC7901 cells and gastric cancer tissues (P = 0.0077, P < 0.01). SGC7901 cells and primary gastric cancer tissues with lymph nodes metastasis had lower miR-100 expression than those of without lymph node metastasis (P = 0.0361, P = 0.0356). The migration ability and invasion of SGC7901 cells transfeced with pre-miR-100 decreased as compared with control cells (P = 0.0025, P = 0.0028 respectively). However, miR-100 expression had no significant effects on the cell proliferation.</p><p><b>CONCLUSIONS</b>Expression of miR-100 inhibits the migration and invasion of gastric cancer cells without significant alteration of proliferation. Therefore, miR-100 may play an inhibitory role in the progression of gastric carcinoma.</p>

Analysis of sex chromosome mosaicisms in early cleavage-stage human embryos and blastocysts with poor embryo quality scores / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze sex chromosome mosaicisms in early cleavage-stage human embryos and blastocysts with poor embryo quality score based on the numbers of pronucleus(PN) zygotes using X,Y dual color fluorescence in situ hybridization (FISH), and to discuss the possible mechanisms.</p><p><b>METHODS</b>Fresh or frozen-thawed early cleavage-stage human embryos and blastocysts with poor embryo quality score not suitable for embryo transfer were studied with dual color FISH.</p><p><b>RESULTS</b>Double signal rate of 2PN among early cleavage-stage embryos was 66.67%, which was significantly higher than 1PN and 3PN embryos. Single signal rate of 1PN early cleavage-stage embryos was 90.41%, which was significantly higher than 2PN and 3PN ones. Three signal rate of 3PN early cleavage-stage embryos was 28.00%, which was significantly higher than 1PN and 2PN ones. Double signal rate of 3PN ones was 46.00%, which was significantly higher than 1PN ones. The polyploid rate of frozen-thawed early cleavage-stage embryos was 23.53%, which was slightly higher than that of fresh embryos, but with no statistical significance. The mosaicism rate of 24 blastocysts was 100.00% and the double signal dominant (≥ 50%) rate was 62.50%, which was significantly higher than the rate of early cleavage-stage embryos.</p><p><b>CONCLUSION</b>Using 2PN as the criterion for embryo quality score cannot guarantee the selection of normal fertilized embryo for transplantation. Frozen-thawed embryos may harbor more polyploid cells. To avoid the selection of embryos with abnormal chromosomes, combinations of pre-implantation genetic diagnosis (PGD) and prenatal diagnosis are necessary. Meanwhile, blastocysts with poor quality scores may provide an important source for embryo stem cells.</p>

THE EFFECT OF hCG STIMULATION GIVEN AT DIFFERENT INTERVIALS ON TESTOSTERONE BIOSYNTHESIS IN CULTURED HUMAN TESTICULAR TISSUE / 中华内分泌代谢杂志

Cultured human testicular tissues were grouped as follows:control group,group tracted with single dosc of human chorionic gonadotropin(hCG), group treated with double doses of hCG given at three-day-interval and group treated with multiple doses of hCG. The dose of hCG was 20kIU/L,and it was added into the medium and incubated for 24 hours. Testosterone was measured by RIA. The result indicated that all three hCG-treated groups had a testosterone secretion peak at 48-72 hours after the first dose of hCG. After the second hCG treatment given in three days interval the testosterone increased again. but its maximal rise reduced. Multiple doses of hCG given successively inhibited the response of Leydig cells to hCG stimulation.