Hemolysis caused by autologous blood transfusion in ectopic pregnancy patient with hemorrhagic shock: a case report / 中国综合临床

Ectopic pregnancy is a common gynecological acute abdomen disease. Once the pregnant tissue is ruptured, it will rapidly develop into hemorrhagic shock or even death. In recent years, blood transfusion from the body is widely used in the rescue of intra-abdominal hemorrhage of ectopic pregnancy, which can reduce the time of cross matching and blood collection, reduce the risk of allogeneic blood transfusion, and enable patients with hemorrhagic shock to receive timely and effective treatment. Hemolysis caused by autologous blood transfusion is rarely reported. Once hemolysis occurs, if it is not handled in time, severe cases can occur acute renal injury, hyperkalemia, or cardiac arrest or even sudden death. We retrospectively analyzed the diagnosis and treatment of a patient with hemolysis after autologous blood transfusion, suggesting that the adverse reactions of blood transfusion occur not only in allogeneic blood transfusion, but also in autologous blood transfusion. It should be handled reasonably in clinical work to reduce the occurrence of similar complications.

Effect of cisplatin combined with 5-fluorouracil on endocrine Ishikawa cells in endometrial cancer and its possible mechanism / 中国生化药物杂志

Objective To investigate the effect of cisplatin and 5-fluorouracil on endocrine Ishikawa cells in the treatment of endometrial cancer. Methods The human endometrial cancer cell line Ishikawa were selected, at the logarithmic growth period, the cells were divided into three groups of 5-fluorouracil (5 mg/L) group, cisplatin (5 mg/L) group and cisplatin (5 mg/L) combined with fluorouracil (5 mg/L) group ( combined group).After treated with corresponding drugs treatment, the cell viability was detected by MTT, the apoptosis was detected by flow cytometry, and the Bcl-2 and p65 expressions cells were detected by Western blot.Results The inhibitory rate of combined group, cisplatin group and 5-fluorouracil group were (41.45 ± 3.13)%, (25.20 ±3.09)% and (23.19 ±4.10)% respectively, and the inhibitory rate in the combined group was significantly higher than that of the other two groups (P<0.05).The apoptosis rate of the combined group, cisplatin group and 5-group were (29.44 ±4.35)%, (5.74 ±1.12)% and (5.82 ±1.78)% respectively, and the apoptosis rate in the combined group was significantly higher than that of the other two groups (P<0.05).The Bcl-2 relative expression in the combined group, cisplatin group and 5-fluorouracil group were (0.31 ±0.11), (1.23 ±0.49) and (1.28 ±0.59), the p65 expression were (0.67 ±0.23), (1.67 ±0.56) and (1.71 ±0.71), combined group of Bcl-2 and p65 relative expressions were obviously less than that of the other two groups ( P <0.05 ) .Conclusion Cisplatin combined with 5-fluorouracil on endocrine Ishikawa cells in the treatment of endometrial cancer could promote cell apoptosis, inhibit the cell proliferation, and its mechanism may be related to the inhibition of Bcl-2 and p65 protein expressions.

Effect of glucose regulated protein 78 on autophagy and apoptosis in ovarian epithelial carcinoma / 中华妇产科杂志

Objective To explore the effect and mechanism of glucose regulated protein 78 (GRP78) on autophagy and apoptosis in ovarian carcinoma, and to investigate the influence on the growth and sensitivity to cisplatin on the ovarian cancer cells.Methods The human ovarian cancer cell line SKOV3 were treated by the GRP78 regulator BAPTA-AM and A23187, which were used to decrease or increase the expression levels of GRP78, respectively.The experiment were divided into three groups.Cells in the group of BAPTA-AM were treated by BAPTA-AM at the final concerntration of 40 μ mol/L for 1 hour.Cells in the group of A23187 were treated by A23187 at the final concerntration of 4 μmol/L for 24 hours.While, cells in the control group were treated by culture medium without any GRP78 regulator for 24 hours.The expressions of GRP78, beclin1, Bcl-2 and CHOP mRNA and protein were detected by reverse transcription (RT)-PCR and western blot.The autophagy levels was observed by green fluorescent protein-microtubule-associated protein 1 light chain 3-Ⅱ (GFP-LC3-Ⅱ) fluorescence staining.The flow cytometry was used to analyse the apoptosis rates of cells.The effect on cell growth and the sensitivity to cisplatin of SKOV3 were accessed by methyl thiazolyl tetrazolium (MTT).Results (1)The mRNA expressions of GRP78, beclin1, Bcl-2 and CHOP in the group of BAPTA-AM were 0.583±0.025, 0.860± 0.055, 0.714±0.032 and 0.811±0.004, respectively.The mRNA expressions of GRP78, beclin1, Bcl-2 and CHOP in the group of A23187 were 0.840± 0.044, 0.654 ± 0.065, 0.908 ± 0.047 and 0.620 ± 0.062, respectively.The mRNA expressions of GRP78, beclin1, Bcl-2 and CHOP in the control group were 0.687± 0.032, 0.772 ±0.029, 0.845 ±0.018, 0.712 ± 0.077, respectively.While the protein expressions of GRP78, beclin 1, Bcl-2 and CHOP in the group of BAPTA-AM were 0.423±0.035, 0.952±0.022, 0.385±0.032, 0.681± 0.095, respectively.The protein expressions of GRP78, beclin1, Bcl-2 and CHOP in the group of A23187 were 0.743 ±0.032, 0.638±0.025, 0.596±0.029, 0.431 ±0.095, respectively.The protein expressions of GRP78, beclin1, Bcl-2 and CHOP in the control group were 0.617±0.031, 0.789±0.083, 0.492±0.036, 0.531 ± 0.003, respectively.The mRNA and protein expressions of beclin and CHOP in the group of BAPTA-AM were both higher than those in the control group (P＜0.05).While, the mRNA and protein expressions of beclin and CHOP in the group of A23187 were both lower than those in the control group (P＜ 0.05).(2) The autophagy fluorescence of SKOV3 in the group of BAPTA-AM, A23187 and the control group were 706±117, 473±128, 595± 126, respectively, in which there were significant differences among three groups (P＜0.05).(3) The apoptosis rate of SKOV3 in the group of BAPTA-AM was (27.4±2.2)％, which was higher than that in the control group [(19.6± 1.4)％, P＜0.05].The apoptosis rate of SKOV3 in the group of A23187 was (12.2± 1.9)％, which was lower than that in the control group (P＜0.05).(4) The comparison of the sensitivity to cisplatin in 3 groups of SKOV3.The 50％ inhibition concentration (IC50) of SKOV3 to cisplatin was (3.02±0.62) mg/L.After treated by BAPTA-AM + cisplatin, the IC50 was (2.00±0.17) mg/L and the sensitivity of SKOV3 to cisplatin was increased by 33.8％, and there was significant difference (P＜0.05), compared with the control group.And after treated by A23187 + cisplatin, the IC50 was (4.91±2.52) mg/L and the sensitivity of SKOV3 to cisplatin was decreasd by 62.6％;and there was significant difference (P＜0.05), compared with the control group.Conclusion GRP78 could regulate autophagy and apoptosis of ovarian cancer cells by regulating the expressions of beclin1, Bcl-2 and CHOP, thereby affecting the sensitivity to cisplatin in ovarian carcinoma, which may be a new method for the treatment and improvement of the sensitivity to cisplatin in ovarian carcinoma.

Effect and mechanism of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The study intended to investigate the effect and mechanism of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma.</p><p><b>METHODS</b>RT-PCR and Western blot were used to test the expression of mTOR and Beclin1 mRNA and protein in ovarian cancer SKOV3 cells after saquinavir induction. MTT assay was used to analyze the influence of saquinavir on cisplatin sensitivity in SKOV3 cells.</p><p><b>RESULTS</b>The IC50 of SKOV3 cells was (5.490 ± 1.148) µg/ml. After induced by Saquinavair 10 µmol/L and 20 µmol/L, the IC50 of SKOV3 cells was increased to (11.199 ± 0.984) µg/ml and (14.906 ± 2.015) µg/ml, respectively. It suggested that the sensitivity of ovarian cancer cells to cisplatin was decreased significantly (P = 0.001). The expression of mTOR and Beclin1 mRNA and protein was significantly different among the five groups: the (Saquinavair+DDP) group of, Saquinavair group, LY294002 group, DDP group and control group (P < 0.001) . The expressions of mTOR and Beclin1 mRNA were highest in the (Saquinavair+DDP) group, 0.684 ± 0.072 and 0.647 ± 0.047, respectively; Secondly, the Saquinavair group, 0.577 ± 0.016 and 0.565 ± 0.037, respectively. The expressions of mTOR and Beclin1 proteins were also highest in the (Saquinavair+DDP) group, 0.624 ± 0.058 and 0.924 ± 0.033, respectively, followed by the Saquinavair group, 0.544 ± 0.019 and 0.712 ± 0.024. 3-MA inhibited the autophagy and restored cisplatin sensitivity in the SKOV3 cells after Saquinavir induced ER stress (P < 0.001).</p><p><b>CONCLUSIONS</b>Saquinavir can effectively induce endoplasmic reticulum stress in SKOV3 cells. Endoplasmic reticulum stress can decrease the sensitivity to cisplatin in SKOV3 cells. The mechanism of the decrease of sensitivity to cisplatin in SKOV3 cells may be that ERS regulates cell autophagy through the mTOR and Beclin1 pathways. ERS of tumor cells and autophagy may become a new target to improve the therapeutic effect of chemotherapy and to reverse the drug resistance in tumor treatment.</p>

Primary study on the signal transduction concerned human papillomavirus 18 E6 oncoprotein / 中华微生物学和免疫学杂志

Objective To explore the relationship of signal transduction among human papillomavirus 18 E6 oncoprotein (HPV18E6), signal transducers and activators of transcription 1 (STAT1), protein kinase R( PKR )/α subunit of eukaryotic initiation factor 2 ( eIF2α ), nuclear factor-kappa Bp65 ( NF-κBp65 ), mitogen-activated protein kinase( MAPK)/c-Jun N-terminal kinase(JNK) ,and possible molecular mechanism. Methods Construct two lentiviral vectors which contain shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and NC sequence( HPV18E6-RNAi-LV, NC-GFP-LV), based on the transduction with HPV18E6-RNAi-LV and NC-GFP-LV into HeLa cell to interfere the expression of HPV18E6 oncogene and NC sequence,the expressions of mRNA and protein( including phosphating patem)of HPV18E6, STATI, PKR, eIF2α, NF-κBp65, MAPK, JNK are measured with RT-PCR and Western blot, the difference of proliferation and sensitivity to carboplatin of HeLa cell are determined with Transwell cell methods and MTT among every groups. Results The expression of HPV18E6 oncogene can affect the expression level of mRNA and protein of NF-κBp65 and PKR genes, also affect phosphating levels of phosphating protein p-STAT1, p-PKR and p-eIF2α;the restraining rates of proliferation and sensitivity to carboplatin of HeLa cell are higher in HPV18E6-RNAi-LV group than the other groups( P＜0. 05 or P＜0.01 ). Conclusion HPV18E6 oncoprotein not only reduces the expression of PKR but dephosphorylates p-STAT1, pPKR and p-eIF2α to restrain activation of PKR/eIF2α signal transduction passage, maintain the proliferation and invading ability of HeLa cell and restrain apoptosis. The signal transduction among HPV18E6, MAPK/JNK are not clear.

Expression and Significance of PKR,p-PKR and p-EIF2α in Human Cervical Lesions / 天津医药

Objective:To identify the relationship between expression of protein kinase R(PKR), phosphating PKR, EIF2α(p-PKR, p-EIF2α) in PKR→EIF2α signal transduction passage and the grades of cervical lesions, the role in generation and progression of cervical tumor and their effects to prognosis of cervical cancer patients. Methods:The expressions of PKR, p-PKR and p-EIF2α in human cervical cancer tissue of 63 cases, cervical intraepithelial neoplasia(CINⅠ-Ⅲ) of 114 cases and normal cervical epithelium of 15 cases were detected by immunohistochemical technique. Results:With the increase in grades of cervical lesions, the positive-expression rate of PKR increased and significantly correlated with the grades of cervical lesions(P < 0.05). With the increase in grades of cervical lesions, the positive-expression rates of p-PKR and p-EIF2α increased firstly, and then decreased. In cervical cancer group, the positive-expression rate of PKR was much higher than that of p-PKR(P < 0.01). The development and progression was quicker in later clinical stages of cervical cancer than that of earlier clinical stages of cervical cancer (P < 0.01). The development and progression of cervical cancer was quicker in patients with negative-expression of p-PKR and p-EIF2α than that in patients with positive-expression of p-PKR and p-EIF2α(P < 0.05). Conclusion:The positive-expression rate of PKR was correlated with the grades of cervical lesions. There are some factors which can impede PKR and EIF2α to be phosphorylated or make p-PKR and p-EIF2α dephosphorylate in high level cervical lesions, which promotes the development and progression of cervical lesions, worsens the prognosis of cervical cancer.

Influence of mifepristone on DNA repair genes and cisplatin sensitivity in human ovarian cancer drug-resistance cells / 中华妇产科杂志

Objective To study the changes of DNA repair genes and enhanced anti-tumor effect of cisplatin induced by mifepristone in human ovarian cancer drug resistance cells.Methods The alterations of cisplatin concentration producing 50％inhibition(IC50)in the COC1/DDP cell lines were examined by methyl thiazolyl tetrazolium(MTT)assay.RT-PCR and flow cytometry were used to analyze the changes of the mRNA of ERCC1,BRCA1,hMLH1 genes and cell cycle and apoptosis.Subcutaneous implantation of COC1/DDP was established in nude mice and the enhanced anti-tumor effect of cisplatin by mifepristone was observed in vivo.ResultsCisplatin IC50 values of COC1/DDP cell were decreased from(3.71±0.38)μg/ml to(3.18±0.46),(1.95±0.14),(0.64±0,18)μg/ml respectively when treated with 2.5,5.0,10.0 μmol/L mifepristone.Mifepristone could down-regulate the mRNA levels of ERCC1,BRCA1,hMLH1 genes and enhance G0/G1 phase block effect pf cisplatin,and 2.5,5.0,10.0 μmol/L mifepristone combined with cisplatin increased rate of cell apoptosis from 0.08％to 5.11％,9.13％and 12.24％ respectively.The percentage of inhibition of xenograft tumor volume in combined treatment group was 70.1％,which was significantly different(P＜0.05).Conclusion By down-regulating ERCC1,BRCA1,hMLH1 genes,blocking G0/G1 phase,and increasing apoptosis rate,mifepristone could enhance anti-tumor effect of cisplatin.

Enhanced cisplatin cytotoxicity by RNA interfering the excision repair cross-complementing 1 gene in ovarian cancer cell lines / 中华妇产科杂志

Objective To study changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing (ERCC) 1 gene in ovarian cancer cell lines. Methods The small interference RNA (siRNA) targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to ovarian cancer cell line ES-2. The mRNA and protein of ERCC1 were evaluated by means of RT-PCR, western blot and immunocytochemistry. The changes of cisplatin sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTT) assay. Results In ES-2 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfection. The sensitivity to cisplatin of ES-2 cell line was increased by 53.88 times after disturbing the ERCC1 gene. Conclusion The sensitivity to cisplatin of ovarian cancer cell lines ES-2 could be enhanced by RNA interfering ERCC1 gene.