Bioinformatics analysis of ferroptosis in spinal cord injury

Ferroptosis plays a key role in aggravating the progression of spinal cord injury(SCI),but the specific mechanism remains unknown.In this study,we constructed a rat model of T10 SCI using a modified Allen method.We identified 48,44,and 27 ferroptosis genes that were differentially expressed at 1,3,and 7 days after SCI induction.Compared with the sham group and other SCI subgroups,the subgroup at 1 day after SCI showed increased expression of the ferroptosis marker acyl-CoA synthetase long-chain family member 4 and the oxidative stress marker malondialdehyde in the injured spinal cord while glutathione in the injured spinal cord was lower.These findings with our bioinformatics results suggested that 1 day after SCI was the important period of ferroptosis progression.Bioinformatics analysis identified the following top ten hub ferroptosis genes in the subgroup at 1 day after SCI:STAT3,JUN,TLR4,ATF3,HMOX1,MAPK1,MAPK9,PTGS2,VEGFA,and RELA.Real-time polymerase chain reaction on rat spinal cord tissue confirmed that STAT3,JUN,TLR4,ATF3,HMOX1,PTGS2,and RELA mRNA levels were up-regulated and VEGFA,MAPK1 and MAPK9 mRNA levels were down-regulated.Ten potential compounds were predicted using the DSigDB database as potential drugs or molecules targeting ferroptosis to repair SCI.We also constructed a ferroptosis-related mRNA-miRNA-lncRNA network in SCI that included 66 lncRNAs,10 miRNAs,and 12 genes.Our results help further the understanding of the mechanism underlying ferroptosis in SCI.

Controlling oxygen content in electro-slag remelting steel by optimizing slag-steel reaction process

The thermodynamic equilibrium of deoxidation reactions between molten slag and steel was calculated using a slag-steel coupling thermodynamic model and the mass conservation model based on the ion-molecular coexistence theory.The study focused on the effects of slag composition and deoxidizer type on the oxygen content of low alloy steel during the electroslag remelting(ESR)process.The measured and predicted values of the oxygen content in remelted ingots,and the contents of FeO and MnO in slags were compared and analyzed.Results show that the measured content of total oxygen has a certain correlation with the trend of dissolved oxygen predicted by the model when using Ca-Si alloys as deoxidizer,but it is not correlated with the trend of dissolved oxygen predicted by the model when using Al as deoxidizer.The deoxidation mechanisms of Ca-Si and Al are different.Ca-Si alloy directly reacts with FeO and MnO in slag to reduce the oxygen potential of slag,hence it can inhibit the transfer of oxygen from the slag to molten steel.While,when Al deoxidizer is used,the oxygen content in steel is mainly reduced through floating up the alumina inclusions.Compared to Al,utilizing Ca-Si alloy as a deoxidizer is more effective in reducing the oxygen content and the amount of inclusions in ESR ingot.

镁造孔多孔NiTi合金的微波烧结制备与表征

为了获得轻质、高强和高阻尼的多孔NiTi合金,采用微波烧结协同镁造孔技术制备多孔NiTi合金。考察多孔NiTi合金的显微组织、力学性能、相变行为、超弹性和阻尼性能。结果表明:当烧结温度低于或等于900℃时,多孔NiTi合金主要由B2 NiTi相和少量B19'NiTi相组成。随着烧结温度的升高,多孔NiTi合金的孔隙率逐渐降低,而压缩强度先增加后下降,在900℃时获得最大值。随着造孔剂Mg的含量从1%增加到7%(质量分数),多孔NiTi合金的孔隙率从37.8%增加到47.1%,而压缩强度从2058 MPa降低至1146 MPa。与碳酸氢铵造孔多孔NiTi合金相比,镁造孔多孔NiTi合金的相变行为发生变化,而压缩强度、超弹性、形状记忆效应和阻尼性能均获得极大的提高。

Perfusion of gastrodin in abdominal aorta for alleviating spinal cord ischemia reperfusion injury

Objective:To observe the effects of perfusion of the gastrodin in abdominal aorta for alleviating the spinal cord ischemia reperfusion injury(SCIRI).Methods:A total of 36 New Zealand white rabbits were divided randomly into sham-operated group(group S),control group(group C) and gastrodin group(group G),12 rabbits for each group.Aorta abdominalis infrarenalis blocking method was applied to establish the SCIRI model.The changes of motor evoked potentials(MEPs) before the ischemia and on 30 min,60 min,6 h,12 h and 24 h of reperfusion of the gastrodin were respectively recorded,and the neurologic function score before the ischemia,on the 6 h,12 h and 24 h of the reperfusion of the gastrodin were assessed.And the changes of the concentration of serum neuron specific enolase(NSE),interleukin(IL)-l β and IL-8 were measured before the ischemia,after 45 min of ischemia,and on 30 min,60 min,6 h,12 h and 24 h of reperfusion of gastrodin.Then the levels of spinal cord nerve cells mitochondrial superoxide dismutase(SOD),reactive oxygen species(ROS),glutathione peroxidase(GSH-PX),malondialdehyde(MDA),total antioxidant capacity(T-AOC) and mitochondrial swelling degree(MSD) were tested and the histopathologic changes in spinal cord tissues were observed.Results:The levels of the NSE,IL-lfter the ischemβ,IL-8,ROS,MDA and MSD of group C were all significantly elevated aia(P<0.01);the levels of the spinal nerve cell mitochondria SOD,GSH-PX and T-AOC were all significantly reduced(P<0.01),MEPs and spinal cord tissue pathology were damaged significantly(P<0.01).The rate of motor neuron abnormalities and the damages of spinal cord tissue pathology of group G were significantly milder than those of group C(P<0.01);the levels of NSE,IL-lROS,MDA and MSD were significantly lower than those of group C(P< 0.01),but tβ,IL-8,he levels of SOD,GSH-PX and T-AOC were all significantly higher than those of group C(P<0.01),and the recovery of neurologic function score during the reperfusion of gastrodin was significantly faster than group C(P<0.01).Conclusions:Perfusion of the gastrodin in abdominal aorta can alleviate the spinal cord ischemiare perfusion injury by promoting the mitochondrial antioxidant capacity and inhibiting the inflammatory reaction.

Digestive tract reconstruction options after laparoscopic gastrectomy for gastric cancer

In addition to the popularity of laparoscopic gastrectomy(LG),many reconstructive procedures after LG have been reported.Surgical resection and lymphatic dissection determine long-term survival;however,the election of a reconstruction procedure determines the postoperative quality of life for patients with gastric cancer(GC).Presently,no consensus exists regarding the optimal reconstructive procedure.In this review,the current state of digestive tract reconstruction after LG is reviewed.According to the determining influence of the tumor site on the procedures of surgical resection and reconstruction,we divide these reconstruction procedures into three categories consistent with the resection procedures.We focus on the technical tips of every reconstruction procedure and examine the surgical outcomes(length of surgery and blood loss)and postoperative complications(anastomotic leakage and stricture)to facilitate gastrointestinal surgeons to understand the merits and demerits of every reconstruction procedure.

Superior Fidelity and Distinct Editing Outcomes of SaCas9 Compared with SpCas9 in Genome Editing

A series of clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPR associated protein 9(Cas9)systems have been engineered for genome editing.The most widely used Cas9 is SpCas9 from Streptococcus pyogenes and SaCas9 from Staphylococcus aureus.However,a comparison of their detailed gene editing outcomes is still lacking.By characterizing the editing outcomes of 11 sites in human induced pluripotent stem cells(iPSCs)and K562 cells,we found that SaCas9 could edit the genome with greater efficiencies than SpCas9.We also compared the effects of spacer lengths of single-guide RNAs(sgRNAs;18–21 nt for SpCas9 and 19–23 nt for SaCas9)and found that the optimal spacer lengths were 20 nt and 21 nt for SpCas9 and SaCas9,respectively.However,the optimal spacer length for a particular sgRNA was 18–21 nt for SpCas9 and 21–22 nt for SaCas9.Furthermore,SpCas9 exhibited a more substantial bias than SaCas9 for nonhomologous end-joining(NHEJ)+1 insertion at the fourth nucleotide upstream of the protospacer adjacent motif(PAM),indicating a characteristic of a staggered cut.Accordingly,editing with SaCas9 led to higher efficiencies of NHEJ-mediated double-stranded oligodeoxynucleotide(dsODN)insertion or homology-directed repair(HDR)-mediated adeno-associated virus serotype 6(AAV6)donor knock-in.Finally,GUIDE-seq analysis revealed that SaCas9 exhibited significantly reduced off-target effects compared with SpCas9.Our work indicates the superior performance of SaCas9 to SpCas9 in transgene integration-based therapeutic gene editing and the necessity to identify the optimal spacer length to achieve desired editing results.

Menopausal Symptoms and Sleep Quality During Menopausal Transition and Postmenopause

Background： Menopausal symptoms and sleep difficulty were physiological processes that were affected by genetic and other factors. This study was to investigate the prevalence of menopausal symptoms and sleep quality in menopausal transition （MT） and postmenopause （PM） women in Taiyuan, Shanxi. Methods： A community-based survey of women＇s menopausal symptoms and sleep quality was conducted between July 2012 and May 2013 at six municipal districts of Taiyuan, Shanxi. A sample of 2429 women aged 40-59 years was divided into four groups： early MT, late MT, early PM, and late PM. Sleep quality in the past 2 weeks before the interview was recorded. The data were analyzed using SPSS 16.0. Results： The prevalence of menopausal symptoms was 49.8%. Mild, moderate, and severe symptoms were observed in 28.9%, 18.5%, and 2.5% of participants, respectively. The highest prevalence of menopausal symptoms occurred in the early postmenopausal stage; the subsequences were the late postmenopausal stage and the early MT stage. Interestingly, among the 13 items of modified Kupperman index, the five most common symptoms were fatigue, arthralgia and myalgia, decreased libido, insomnia, and nervousness. Meanwhile, 55% perimenopausal women had poor sleep. Conclusions： Menopausal symptoms are common but mild among women in Taiyuan, Shanxi during MT and PM. In these stages, the prevalence of poor sleep is high.

Effect of Long Protocol on the Outcome of In Vitro Fertilization-embryo Transfer in Patients with Endometriosis Compared with Prolonged Protocol

Objective To investigate the effect of different down-regulation protocol on the in vitro fertilization-embryo transfer （IVF-ET） outcomes in infertile patients with endometriosis （EMs）.Methods A retrospective case control study was performed. Totally 294 infertile patients with EMs were enrolled. And 109 patients （116 cycles） received prolongedprotocol as the control, 185 patients （193 cycles） received long protocol as case group, all followed by standard controlled ovarian hyperstimulation （COH）. Response to gonadotropins, the fertilization rate, the cleavage rate, the implantation rate, the clinical pregnancy rate and the miscarriage rate were measured and analyzed between the two groups. Results A trend toward better ovarian response was observed in long protocol group. Higher fertilization rate, lower total dose of rFSH, shorter duration of stimulation and more endometrial thickness on the day of hCG injection were observed in long protocol group compared with those of prolonged protocol group, and the difference was significant （P〈0.05）. In addition, the clinical pregnancy rate, the cleavage rate and the implantation rate also had an increase trend in long protocol group compared with those of prolonged protocol group, but without significant differences. Conclusion Long protocol regimen before IVF-ET in patients with EMs resulted in a trend toward better ovarian response and higher clinical pregnancy rates than prolonged protocol regimen did.

HDAC inhibitors improve CRISPR-mediated HDR editing efficiency in iPSCs

Genome-edited human induced pluripotent stem cells(iPSCs)hold great promise for therapeutic applications.However,low editing efficiency has hampered the applications of CRISPR-Cas9 technology in creating knockout and homology-directed repair(HDR)-edited iPSC lines,particularly for silent genes.This is partially due to chromatin compaction,inevitably limiting Cas9 access to the target DNA.Among the six HDAC inhibitors we examined,vorinostat,or suberoylanilide hydroxamic acid(SAHA),led to the highest HDR efficiency at both open and closed loci,with acceptable toxicity.HDAC inhibitors equally increased non-homologous end joining(NHEJ)editing efficiencies(~50%)at both open and closed loci,due to the considerable HDAC inhibitor-mediated increase in Cas9 and sgRNA expression.However,we observed more substantial HDR efficiency improvement at closed loci relative to open chromatin(2.8 vs.1.7-fold change).These studies provide a new strategy for HDRediting of silent genes in iPSCs.

Potential mechanism of cholagogic effect about Gardenia Jasminoides Ellis(Zhizi)-mediated increase of bile acids urinary excretion in normal rats

Objectives： Gardenia Jasminoides Ellis（Zhizi）, belonging to Rubiaceae family, has been traditionally used for treatment cholestasis and jaundice for centuries in Asian countries. In the theory of Traditional Chinese Medicines, Zhizi could dispel dampness and heat via the urine to execute its choleretic effects. However, the potential molecular mechanism has been still poorly clarified. Here, we investigated the effect of different dose of Zhizi aqueous extract powder（0.3 g/kg/d and 0.9 g/kg/d） on urinary excretion of bile acids（BAs）, and defined the potential mechanism via renal BAs efflux transporters Mrp2 and Mrp4 in normal rats.Methods： Male Wistar rats were orally administrated with 0.3 or 0.9 g/kg/d dose of Zhizi aqueous extract powder for 2 weeks, then body weight, serum aminotransferase, total BAs concentrations in liver, bile,serum, kidney and urine, 1 h bile flow, 12-h urinary volume, biliary and urinary excretion amount of total BAs as well as protein expression of major renal BAs efflux transporter Mrp2 and Mrp4, were all evaluated.Results： Zhizi especially the high dose of Zhizi aqueous extract powder could reduce hepatic total BAs concentration. Additionally, bile flow and biliary excretion had no significant difference, but the remarkable increasing urinary excretion of BAs and 2 to 3 folds up-regulated renal Mrp2 expression were observed after administrated with Zhizi as compared with the control group.Conclusion： The findings indicate that Zhizi reduces hepatic total BAs level by increasing urinary excretion rather than the biliary excretion of BAs, which, in turn ascribed to elevated protein expression of Mrp2 at apical membrane of renal tubular epithelial cells.

Mifepristone (RU486) Inducing Abortion in a Mouse Model by Regulating Innate and Adaptive Immune Responses

Objective:Mifepristone(RU486),one of the most common medications for artificial abortion,attenuates the immunoregulatory effects of progesterone.However,the specific immune regulatory mechanism of RU486 in abortion remains unknown.We intended to investigate the immunomodulatory effects of RU486 on abortion.Methods:Sixty female mice were divided into the control group(0 mg RU486)and RU486 group(2 mg/kg RU486).The uterus,peripheral blood,and spleen were obtained for isolation of specific cell types.The population and phenotype of immune cells in the decidua,peripheral blood,and spleen were analyzed using flow cytometry.Statistical differences between groups were determined using two-tailed t-test.For all statistical tests,P<0.05 was considered statistically significant.Results:RU486 effectively induced abortion in pregnant mice,with a significantly higher number of decidual macrophages(dMφ)(control group=25.55%±2.467%,RU486 group=19.41%±1.423%;P<0.05),especially the major histocompatibility complex II high subset.No difference in Mφnumber was observed in the spleen or peripheral blood.Moreover,the dMφfrom mice with RU486-induced abortion displayed a remarkable activated phenotype,with increased expressions of inducible nitric oxide synthase,tumor necrosis factor-α,and interleukin(IL)-12 but decreased expressions of arginase-1 and IL-10.We also found elevated levels of decidual CD4+T-cells in the RU486 group that exhibited a higher level of the proinflammatory cytokine interferon-γand a lower level of the anti-inflammatory cytokines,IL-4 and IL-10.Conclusions:We report a new mechanism of RU486-induced abortion via the regulation of innate cell Mφactivation and the adaptive response of CD4+T-cells present in the decidua but not the periphery.

Triplet excitation dynamics of β-carotene studied in three solvents by ns flash photolysis spectroscopy

Upon anthracene-sensitizing, triplet excitation dynamics of β-carotene（β-Car） were studied in nhexane, in methanol, and in acetonitrile, respectively, by ns flash photolysis spectroscopy. In n-hexane,only the bleaching of the ground state absorption（GSB） and the excitation triplet（^3Car＊） absorption were observed, and there were no cationic species detected. In both methanol and acetonitrile, similar excitation dynamics were observed, i.e.,^3Car＊ having a similar lifetime to that in n-hexane, and the immediate generation of the cation dehydrodimer（^#[Car]2^＋） upon excitation following transformation into the radical cation Car＊^＋, since Car＊^＋ has much longer lifetime in acetonitrile than in methanol. The results prove that both solvent and carotenoid structure determine the triplet excitation mechanism.

Difference between 2 h and 3 h 75 g glucose tolerance test in the diagnosis of gestational diabetes mellitus(GDM):Results from a national survey on prevalence of GDM

The possibility of the 2 h oral glucose tolerance test(OGTT)as an alternative to the 3 h OGTT was investigated based on data from a national survey on pregnancy-associated diabetes.Data were retrieved from 4179 pregnant women who had OGTT performed after an abnormal 50 g glucose challenge test(GCT).All of the 4 glucose levels during their OGTT were collected and analyzed.According to American Diabetes Association(ADA)gestational diabetes mellitus(GDM)diagnostic criteria,among the 4179 pregnant women who required OGTT,3429(82.1%)were normal and 750(17.9%)were diagnosed as GDM.If the 3rd h glucose levels were omitted from OGTT,79 cases of GDM(10.5%)would be overlooked.No trend was shown where women with more risk factors were more likely to be overlooked if the 3rd h test was omitted(χ^(2) for trend=0.038,P>0.05).No significant differences were found in the rate of cesarean section(CS),preterm births or macrosomia between the 79 cases and those with normal OGTT results and in the gestational weeks when OGTT was performed.It shows that in order to diagnose one woman with GDM,another 52 pregnant women would have an innocent 3rd h glucose test.Omission of the 3rd h glucose test in OGTT might be reasonable due to its convenience,better compliance and a small number of possibly miss-diagnosed cases,and their pregnancy outcomes have no significant difference from those of normal pregnant women.