Fabrication method for a magnetically induced highly oriented nanohydroxyapatite/collagen composite.

Both collagen fibres and nanohydroxyapatite crystals have anisotropic magnetisation, which allows them to be oriented by a high magnetic field. Highly oriented nanohydroxyapatite/collagen composites were prepared using a high magnetic field combined with in situ synthesis. These highly oriented composites were investigated and compared with conventional composites. The results showed that the collagen fibres in the magnetically induced highly oriented nanohydroxyapatite/collagen composites had a preferred orientation and smaller molecular spacing, while the nanohydroxyapatite crystals were tightly adhered along the collagen fibre surface. The magnetically induced composites exhibited superior resistance to swelling and degradation along with high compressive strength. This artificial composite, with a structure and composition similar to natural bone, represents a new idea for improving materials for vertical or horizontal bone augmentation.

Bioinformatics analysis of m6A-associated genes in steroid-induced osteonecrosis of the femoral head / 中国组织工程研究

BACKGROUND:m6A modification has been confirmed to play an important role in the occurrence and development of osteonecrosis of the femoral head;however,the role of m6A modification patterns in steroid-induced osteonecrosis of the femoral head remains unknown. OBJECTIVE:Bioinformatics analysis was performed based on the Gene Expression Omnibus(GEO)database to analyze the differential expression of the m6A gene in steroid-induced osteonecrosis of the femoral head,predict the downstream targeted miRNAs,and investigate the potential pathogenesis. METHODS:Expressing profiles of mRNA data of steroid-induced osteonecrosis of the femoral head were downloaded from GEO database(GSE123568).Differentially expressed genes(DEGs),Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using the R software.After obtaining these differentially methylated m6A genes(m6A-DEGs),we analyzed GO function and KEGG pathway enrichment and compared the correlation among the m6A-DEGs typing according to gene expression.The protein-protein interaction network and core gene subnetwork of m6A-DEGs were constructed using Cytoscape software.The m6A-DEGs-associated potential miRNAs were predicted using the TargetScan,miRTarBase,and miRBD databases.Simultaneously,ChIPBase and hTFtarget databases were used to predict potential transcription factors of seven core genes,then m6A-miRNA and transcription factor-m6A regulatory networks were constructed separately.Finally,the expression levels of the seven core m6A-DEGs were verified by using the GSE74089 dataset. RESULTS AND CONCLUSION:(1)A total of 2 460 common DEGs were screened out from datasets,among which 1 455 genes were upregulated and 1 005 genes were downregulated.(2)A total of 14 m6A-DEGs were identified in the datasets.Among them,11 m6A-DEGs were up-regulated and 3 m6A-DEGs were down-regulated.Differential gene expression was considered significant for m6A-DEGs in steroid-induced osteonecrosis of the femoral head(P<0.05).Spearman correlation analysis showed a significant correlation between m6A-DEGs.(3)GO and KEGG enrichment analysis showed that m6A-DEGs were mainly enriched in myeloid cell differentiation and development,immune and cytokine receptor activity,osteoclast differentiation,AMPK signaling pathway and interleukin-17 signaling pathway.(4)The seven core genes of m6A-DEGs contained YTHDF3,YTHDF1,YTHDF2,ALKBH5,METTL3,HNRNPA2B1,and HNRNPC.A total of 44 miRNAs overlapping were detected in the miRTarBase,miRDB,and TargetScan databases.Totally 79 transcription factors overlapping were found in the ChIPBase and hTFtarget databases.(5)The expression levels of six core m6A-DEGs in the GSE74089 dataset were consistent with those in the GSE123568 dataset.(6)These findings confirm that the seven m6A-DEGs identified through bioinformatics techniques play a regulatory role in the expression of various miRNAs,transcription factors,AMPK,and interleukin-17 signaling pathways,and these genes have a significant impact on the differentiation and development of bone marrow cells as well as osteoclast differentiation in steroid-induced osteonecrosis of the femoral head.Consequently,these findings offer data support and establish a research direction for future investigations into the pathogenesis and targeted therapeutic strategies for steroid-induced osteonecrosis of the femoral head.

The effects of repeated high acceleration on implant osseointegration in SD rats / 实用口腔医学杂志

Objective:To investigate the effects of repeated high acceleration(+Gz)on implant osseointegration in SD rats.Methods:18 SD rats were divided into+Gz and control groups randomly(n=9),and 1 implant was placed in each femur of the rat's lower limb.24 hours postoperatively,the experimental rats were exposed to+Gz of 4 to 9 G with 1 G/s environment 3 times a week,while the con-trol rats were fed normally.3 rats from each of the 2 groups were sacrificed at 2,4 and 8 weeks after implantation.Micro-CT,sequential fluorescence double labeling,and histological examination were perfomed for the analysis of implant osseointegration.Results:The bone volume fraction(BV/TV),trabecular number(Tb.N),mineral apposition rate(MAR),implant-bone contact rate(BIC)and bone area in implant thread(BA)of the+Gz group were significantly lower than those of the control group at 2 weeks(P＜0.05),and so as to MAR,BA at 4 weeks(P＜0.05),while there was no significant difference of the parameters at 8 weeks after implantation.Conclusion:In SD rats early exposure to+Gz environment postoperatively may have a negative effect on initial osseointegration by slowing bone forma-tion.However,it will not lead to poorer bone mass when sustained over a long period.

LAMP3 inhibited the proliferation,metastatic and PC-3-induced vasculogenesis of HUVEC by regulating VEGF/AKT signaling / 实用医学杂志

Objective To explore the impactof Lysosome-Associated Membrane Protein 3(LAMP3)on theproliferation,migration and angiogenesis of PC-3 cells.Methods LAMP3 expression in normal prostate epithelial cells and prostate cancer bone metastasis cells was detected using western blot and RT-PCR.Stable LAMP3-silenced PC-3 cells were constructed,and the effects of LAMP3 on proliferation,invasion,and migration of PC-3 cells were assessed using CCK8,scratch assay,and transwell assay,respectively.ELISA and angiogenesis assays were employed to examine the expression of VEGF and MMP9,as well as angiogenesis of HUVEC cells induced by PC-3 cells.Finally,WB and RT-PCR were used to detect the expression of VEGF,AKT/p-AKT.Results Our findings showed that the expression level of LAMP3 was significantly higherin prostate cellsthan in normal prostate epithelial cells,especially in PC-3 cells(P<0.05).We also found that silencing LAMP3 could inhibit the proliferation,migration and invasion of PC-3 cells,along with the expression of VEGF and MMP9 and the PC-3 cells-induced angiogenesis,and these results were statistically significant(P<0.05).Furthermore,LAMP3 downregulated the expression of VEGF and AKT/p-AKT in PC-3 cells.Conclusion LAMP3 can affect the proliferation,migrationand angiogenesis of PC-3 cells through the regulation of VEGF/AKT pathway.Thus,LAMP3 might be a potential thera-peutic target for prostate cancer bone metastasis.

Immunogenicity of BCG-CpG-DNA adjuvant rabies vaccine(MRC-5 cell)for human use in cynomolgus monkeys / 中国人兽共患病学报

To evaluate the immunogenicity of BCG-CpG-DNA-adjuvanted human rabies vaccine(MRC-5 cell)in cynomol-gus monkeys,we randomly divided monkeys into a control group,three-dose group,and four-dose group.The three-dose and four-dose groups were vaccinated with BCG-CpG-DNA-adjuvanted human rabies vaccine for human use at 0,7,and 21,or at 0,3,7,and 14 days,respectively.The control group was vaccinated with a commercially available domestic vaccine with a 2-1-1 protocol.Serum samples were collected from all monkeys before(0 day)and after immunization,on days 7,14,28,35,49,and 63.RFFIT and ELISPOT were used to detect anti-rabies virus neutralizing antibodies and cytokines(IFN-γ and IL-2).The anti-rabies virus neutralizing antibodies in all groups were negative before immunization.The three-dose and control groups showed an increasing trend from 7 to 28 days after primary immunization,and the highest level was reached on the 28th day;in contrast,the four-dose group showed a peak on the 14th day.On the 28th day,the neutralizing antibody levels in the control and three-dose groups were significantly higher than those in the four-dose group.On the 35th day after the initial immuniza-tion,the neutralizing antibodies in the three-dose group was significantly higher than those in the four-dose group.The levels of IL-2 and IFN-γ in peripheral blood lymphocytes in the three-dose and four-dose groups tended to increase from 7 to 14 days and peaked on the 14th day;the level of IFN-γ in the three-dose group was significantly higher than that in the control group on the 14th day.Otherwise,no significant differences in the levels of neutrali-zing antibodies and cytokines were observed among all groups at all other time points.Thus,BCG-CpG-DNA adjuvanted human rabies vaccine(MRC-5 cell)has excellent immunogenicity and application value for optimizing immunization procedures.

Mechanism of ginkgolide B antagonizing vascular endothelial injury by inhibiting endoplasmic reticulum stress / 实用医学杂志

Objective To investigate the potential of ginkgolide B(GB)in mitigating vascular endothelial injury by antagonizing endoplasmic reticulum stress(ERS)and elucidate its underlying molecular mechanism.Methods An injury model of human bone marrow-derived endothelial progenitor cells(EPCs)induced by tunica-mycin(TM)was established.Cell proliferation was assessed using MTS assay,while cell viability was determined through Calcein-AM/EthD-I double staining.Transwell assay was employed to evaluate cell migration ability.DCFH-DA staining was utilized to measure intracellular ROS levels,and NADPH activity was quantified via ELISA.JC-1 and DiOC6 staining were performed for qualitative and quantitative assessment of mitochondrial membrane potential respectively.Qrt-pcr analysis was conducted to determine mRNA expression levels,whereas western blot analysis enabled detection of protein expression levels in the cells.Results GB dose-dependently attenuated tunicamycin-induced ERS-mediated endothelial injury in hEPCs,as evidenced by decreased cell viability,impaired cell migration,and angiogenesis inhibition(P<0.01).Furthermore,GB treatment significantly reduced ROS production and NADPH levels within the cells(P<0.01),while also inhibiting ERS-mediated decline in mitochondrial membrane potential concentration-dependently(P<0.01).Additionally,GB inhibited the expression of ERS-related proteins such as GRP78,ATF4,CHOP etc.,regulated apoptosis-related protein Bcl-xl,Bax cleaved caspase-4 cytochrome c;thereby effectively counteracting endoplasmic reticulum stress-induced cellular damage.Conclusions GB exerts a protective effect on vascular endothelium by antagonizing endoplasmic reticulum stress;this mechanism may be attributed to its ability to reduce intracellular reactive oxygen species levels.It also suppresses the expression of ERS-related proteins(CHOP78 and ATF4),and modulates apoptosis-associated proteins(Bcl-xl,Bax,cleaved caspase-4,and cytochrome c).

Thalassemia Gene Detection Results and Application Value of Hematological Indexes Among Pregnant Women in Xindu District of Chengdu City / 中国实验血液学杂志

OBJECTIVE@#To analyze the genotype characteristics of α- and β-thalassemia and the diagnostic value of hematological indexes in pregnant women in Xindu District of Chengdu.@*METHODS@#The blood routine parameters(MCV) <80 fl and (or) (MCH) <27 pg and hemoglobin electrophoresis were used to screen the pregnant women, PCR-reverse dot blot hybridization(PCR-RDB) technique was used to detect the common α- and β-thalassemia gene types in the primary screening positive population. The husbands of the diagnosed pregnant women were recalled for gene testing, and the highly suspected patients were checked by gene sequencing.@*RESULTS@#Among the 7 049 pregnant women, 1 740(24.68%) cases were positive for primary screening. 180 patients were diagnosed as thalassemia gene positive, among them, 94 cases (52.22%) of α-thalassemia were detected and six genotypes were found, in which --SEA /αα genotype was the highest (58 cases, 61.70%); 82 cases (45.56%) of β-thalassemia were detected and ten genotypes were found while CD41-42/N and CD17/N genotypes were the most common; there were 4 cases(2.22%) with α combined with β-thalassemia. Through clinical follow-up survey, there were 4 couples with the same type of thalassemia, one of them was induced labor after diagnosis of hemoglobin H disease. Receiver operating curve (ROC curve) was used to analyze the diagnostic value of hematological parameters in thalassemia positive pregnant women. The results showed that AUC(HBA2)<AUC(MCHC)<AUC(RDW-SD)<AUC(MCH)<AUC(MCV) (P<0.01).@*CONCLUSION@#The most common genotypes of α- and β-thalassemia in pregnant women in Xindu District of Chengdu were --SEA /αα, CD41-42/N, CD17/N. The blood routine indicators (HBA2、RDW-SD、MCHC、MCH、MCV) have high diagnostic value for screening of thalassemia.

Preliminary Study on Screening and Identification of Lewis a Antigen Mimic Epitope in Alpaca Phage Display Nanobody Library / 中国实验血液学杂志

OBJECTIVE@#To establish a new method for synthesizing Lewis blood group antigens, that is, the mimotopes of Lewis blood group antigens were screened by using an alpaca phage display nanobody library.@*METHODS@#We selected mimotopes of the Lewis a (lea) antigen by affinity panning of an alpaca phage display nanobody library using a monoclonal anti-lea antibody. Enzyme-linked immunosorbent assay (ELISA) was used to test the affinity of the positive clones for the monoclonal anti-lea antibody, and the high-affinity positive clones were selected for sequencing and synthesis. Finally, the sensitivity, specificity and reactivity of the synthesized lea mimotope in clinical samples were verified by ELISA.@*RESULTS@#A total of 96 phage clones were randomly selected, and 24 were positive. Fourteen positive clones with the highest affinity were selected for sequencing. The result showed that there were 5 different sequences, among which 3 sequences with the highest frequency, largest difference and highest affinity were selected for expression and synthesis. The sensitivity and specificity of lea mimic antigen by ELISA showed that, the minimum detection limit of gel microcolumn assay (GMA) and ELISA method were 25 times different, and the lea mimic antigen had no cross reacted with the other five unrelated monoclonal antibodies(P<0.001). Finally, 30 clinical plasma samples were analyzed. The mean absorbance of the 15 positive plasma samples was significantly higher than that of the 15 negative plasma samples (P=0.02). However, the positive signal values of the clinical samples were much lower than those of the monoclonal antibodies.@*CONCLUSION@#A new method of screening lea mimic antigen by using alpaca phage nanoantibody library has been established, which is expected to realize the screening of lea mimotopes, thus realizing the application of high-sensitivity detection methods such as ELISA and chemiluminescence in blood group antibody identification.

Diterpenoid alkaloids from roots of Aconitum kongboense / 中国中药杂志

The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).

Preconditioning in lowered oxygen enhances the therapeutic potential of human umbilical mesenchymal stem cells in a rat model of spinal cord injury.

Human umbilical cord mesenchymal stem cells (UCMSCs) have recently been shown to hold great therapeutic potential for the treatment of spinal cord injury (SCI). However, the number of engrafted cells has been shown to decrease dramatically post-transplantation. Physioxia is known to enhance the paracrine properties and immune modulation of stem cells, a notion that has been applied in many clinical settings. We therefore hypothesized that preconditioning of UCMSCs in physioxic environment would enhance the regenerative properties of these cells in the treatment of rat SCI. UCMSCs were pretreated with either atmospheric normoxia (21% O2, N-UCMSC) or physioxia (5% O2, P-UCMSC). The MSCs were characterized using flow cytometry, immunocytochemistry, and real-time polymerase chain reaction. Furthermore, 10(5) N-UCMSC or P-UCMSC were injected into the injured spinal cord immediately after SCI, and locomotor function as well as cellular, molecular and pathological changes were compared between the groups. We found that N-UCMSC and P-UCMSC displayed similar surface protein expression. P-UCMSC grew faster, while physioxia up-regulated the expression of trophic and growth factors, including hepatocyte growth factor (HGF), brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor(VEGF), in UCMSCs. Compared to N-UCMSC, treatment with P-UCMSC was associated with marked changes in the SCI environment, with a significant increase in axonal preservation and a decrease in the number of caspase-3+ cells and ED-1+ macrophages. These changes were accompanied by improved functional recovery. Thus, the present study indicated that preculturing UCMSCs under 5% lowered oxygen physioxic conditions prior to transplantation improves their therapeutic potential for the treatment of SCI in rats.

Study on anti-inflammatory effect of Salvia miltiorrhiza-Kushen herbpairs extract based on network pharmacology / 中国药理学通报

Aim To predict the potential targets of Salvia miltiorrhiza- Kushen herb pairs extracts (SK) and explore its anti-inflammatory effect on modelsbased on network pharmacology, so as to provide the research foundation of both new drugs and anti-inflammatory mechanism. Methods Network pharmacology was used to predict the potential anti-inflammatory targets of SK. Ear edema model was used to study the anti-in- flammatoiy effects of SK. Luminex liquid-phase chip analysis technology was used to observethe changes in secretion of pro-inflammatory cytokines in peripheral serumafter transdermal administration in mice by SK. Intraperitoneal injection of Evans blue experiment was used to simulate the exudation of inflammatory factors, and the effect of SK on capillary permeability in mice- was explored. Results Network pharmacology was used to predict that the anti-inflammatory effect of SK- was mainly related to immune-related processes and VEGF signaling pathways, unravelingthe relationship between the anti-inflammatory mechanismand the decrease of pro-inflammatory factors and vascular permea- bility. Conclusions The experiments verify that the results are the same as the prediction by network pharmacology. The anti-inflammatory effect involves decline the IL-6 levels, granulocyte colony-stimulating factor( G-CSF) levels and vascular permeability in the STAT3 pathway. Asthemain components of SK, tanshi- none, matrine and oxymatrine are linked to anti-in- flammatory effects.

Predictive value of cerebroplacental ratio for perinatal outcomes of induction of labor in prolonged pregnancy / 中华围产医学杂志

Objective:To investigate the predictive value of cerebroplacental ratio (CPR) for adverse perinatal outcomes of induction of labor in prolonged pregnancy.Methods:This retrospective study recruited 315 singleton pregnant women who had induced labor due to prolonged pregnancy (≥41 gestational weeks) in the First Affiliated Hospital of Chongqing Medical University from January 1, 2019 to April 30, 2020. Based on the occurrence of adverse perinatal outcomes (emergency delivery due to persistent abnormal fetal heart rate monitoring, umbilical artery blood pH at birth <7.2, 5 min Apgar scores<7, transferring to neonatal intensive care unit after birth, chorioamnionitis and vaginal delivery converted to cesarean section), they were divided into two groups: case group ( n=76) and normal group ( n=239). Clinical features and umbilical artery blood flow, middle cerebral artery (MCA) flow and CPR measured in the last ultrasound scan before induction were compared between the two groups using student's t-test, Mann-Whitney U test and Chi-square test. Receiver operating characteristic (ROC) curve was used to analyze the predictive values of umbilical artery blood flow, MCA flow and CPR for the adverse perinatal outcomes. Multivariate logistic regression analysis was used to screen the meaningful predictors. Results:Compared with the normal group, the umbilical artery pulsatility index (PI) (0.9±0.1 vs 0.8±0.1, t=-5.458, P<0.001) and the percentage of abnormal CPR (<1.0) increased significantly [21.1%(16/76) vs 6.3%(15/239), χ2=14.190, P<0.001] in the case group, while the MCA-PI and CPR decreased significantly (1.1±0.2 vs 1.3±0.3, t=5.658, P<0.001; 1.2±0.3 vs 1.6±0.5, t=8.940, P<0.001). The areas under the ROC curves of umbilical artery PI, MCA-PI and CPR for predicting adverse perinatal outcomes were 0.71, 0.71 and 0.77, respectively. CPR had the highest sensitivity (0.74) compared with umbilical artery PI (0.68) and MCA-PI (0.71), but the specificity of them were similar (0.67, 0.66 and 0.66). Multivariate logistic regression analysis showed that only CPR was the independent risk factor for adverse perinatal outcomes ( OR=0.028, 95% CI: 0.010-0.080, P<0.001). Conclusions:As an indicator for early prediction of adverse perinatal outcomes of induction of labor in prolonged pregnancy, CPR was more sensitive but less specific.

Canagliflozin alleviates renal fat deposition and improves renal oxygenation levels in patients with newly-diagnosed type 2 diabetes mellitus / 中华内分泌代谢杂志

Objective:To evaluate the effect of canagliflozin on intrarenal fat content and oxygenation in newly-diagnosed type 2 diabetes patients.Methods:Twenty-three newly-diagnosed type 2 diabetes patients were divided into canagliflozin( n=11) and glimepiride control( n=12) groups .Both groups received MRI scanning with Dixon MRI and BOLD MRI sequence to assess patients′ intrarenal fat content, oxygenation level before treatment and 24 weeks after treatment. Fasting blood glucose, glycosylated hemoglobin, blood uric acid, blood lipids, blood pressure, weight, and other metabolic index were also tested before and after treatment. Furthermore, the relationship between body mass index(BMI) and intrarenal fat content and the correlation between changes in intrarenal fat content and improvement in renal hypoxia were analyzed. Results:No significant differences were found in baseline age, body weight, fasting blood glucose, glycosylated hemoglobin, blood lipid, and serum uric acid between the two groups. There was no significant difference in fasting blood glucose, glycosylated hemoglobin, cholesterol(CHO), low-density lipoprotein-cholesterol(LDL-C), and triglycerides(TG) levels in both groups after 12 and 24 weeks of treatment. The decrease in body weight, blood uric acid level, and diastolic blood pressure from baseline in the canagliflozin group was greater than those in the control group( P<0.05). Two groups of patients with type 2 diabetes at baseline had no obvious difference in intrarenal fat content, and the patients′ BMI showed no obvious correlation with degree of intrarenal fat accumulation. Canagliflozin treatment for 24 weeks could reduce intrarenal fat content, which was higher than that of control group. The R2 * values of renal cortex and medulla in the canagliflozin group decreased from baseline by 19.22% and 22.63% respectively( P<0.05), whereas no significant difference was seen in the glimepiride control group. The decrease of intrarenal fat content in the canagliflozin group was related to the improvement of renal cortex and medulla oxygenation. Conclusion:Canagliflozin can reduce intrarenal fat accumulation and improve renal cortical hypoxia in newly diagnosed type 2 diabetes patients with normal renal function.

Phosphorylation of Cofilin-1 Enhances Paclitaxel Resistance of Epithelial Ovarian Cancer Cells by Inhibiting Apoptosis / 生物医学与环境科学（英文）

Objective@#To investigate the molecular mechanism of high phosphorylation levels of cofilin-1 (p-CFL-1) associated with paclitaxel resistance in epithelial ovarian cancer (EOC) cells.@*Methods@#Cells displaying varying levels of p-CFL-1 and CFL-1 were created by plasmid transfection and shRNA interference. Cell inhibition rate indicating paclitaxel efficacy was assessed by Cell Counting Kit-8 (CCK-8) assay. Apoptosis was assessed by flow cytometry and protein levels were detected by western blotting. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of phosphokinases and phosphatases of CFL-1. Survival analysis evaluated the correlation between the prognosis of EOC patients and the levels of p-CFL-1 and slingshot-1 (SSH-1).@*Results@#High levels of p-CFL-1 were observed in EOC cells that survived treatment with high doses of paclitaxel. SKOV3 cell mutants with upregulated p-CFL-1 showed impaired paclitaxel efficacy, as well as decreased apoptosis rates and pro-survival patterns of apoptosis-specific protein expression. Cytoplasmic accumulation of p-CFL-1 inhibited paclitaxel-induced mitochondrial apoptosis. SSH-1 silencing mediated CFL-1 phosphorylation in paclitaxel-resistant SKOV3 cells. Clinically, the high level of p-CFL-1 and the low level of SSH-1 in EOC tissues were closely related to chemotherapy resistance and poor prognosis in EOC patients.@*Conclusion@#The SSH-1/p-CFL-1 signaling pathway mediates paclitaxel resistance by apoptosis inhibition in EOC and is expected to be a potential prognostic predictor.

Optimal administration time of indomethacin in preventing hyperamylasemia after ERCP and reducing pain degree / 中华胰腺病杂志

Objective:To observe the effects of different indomethacin anal suppository administration time on increased serum amylase, pancreatitis onset and the degree of pain after ERCP, and explore the optimal administration time.Methods:A total of 240 patients with cholangiopathy who were admitted and planned to be treated by ERCP in Department of General Surgery of Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from May 2018 to May 2019 were randomly divided into 6 groups using random number method with 40 patients in each group, who were given 100 mg intrarectal indomethacin at 120 min, 60 min, 30 min before ERCP and 30 min, 60 min, 120 min after ERCP. The incidence of post-ERCP hyperamylasemia, post-ERCP pancreatitis (PEP) and the degree of postoperative pain were compared among different groups.Results:The increased times of blood amylase level and the incidence of hyperamylasemia and PEP at 24 hour, and postoperative 3 h and 24 h pain scores in preoperative medication group were significantly lower than those in postoperative medication group [(4.2±1.9) vs(4.7±2.1), 36.7% vs 46.7%, 19.2% vs 24.2%, (5.9±2.1) vs(6.4±1.4) and(3.8±1.7) vs (4.3±1.4)score, all P<0.05), and the amylase increased times (2.5±1.2), the incidence of hyperamylasemia and PEP[(27.5%(11/40) vs 12.5%(5/40), and postoperative pain score (3.0±1.2) were the least in preoperative 30 min medication group. Conclusions:Indomethacin anal suppository can reduce the incidence of postoperative hyperamylasemia and PEP and mitigate the pain degree after ERCP, and indomethacin given 30 minutes before ERCP may obtain the best preventive effect.

The predictive value of cerebroplacental ratio in stageⅡof twin-twin transfusion syndrome / 中华超声影像学杂志

Objective:To analyze the cerebroplacental blood flow distribution characteristics in monochorionic-diamniotic(MCDA) twin pregnancies with stageⅠ-Ⅱof twin-twin transfusion syndrome (TTTS), and investigate the predictive value of cerebroplacental ratio (CPR) in the stageⅡ of TTTS.Methods:The cerebroplacental blood flow distribution were analyzed retrospectively in 68 cases from June 2017 to June 2019 in the First Affiliated Hospital of Chongqing Medical University (34 cases for each stage) with TTTS, including the umbilical artery pulsatility index (UA-PI), middle cerebral artery peak systolic velocity (MCA-PSV), middle cerebral artery pulsatility index (MCA-PI), CPR, and their discordances(UA-PI disc, MCA-PSV disc, MCA-PI disc and CPR disc). Results:The differences of UA-PI, MCA-PI, and CPR between donors and recipients in TTTS Ⅱ were statistically significant (all P<0.05), the differences of UA-PI disc, MCA-PI disc and CPR disc between TTTS Ⅰ and TTTS Ⅱ were statistically significant (all P<0.05). Multivariate Logistic regression analyses showed that only CPR disc was independently associated with the stage Ⅱ of TTTS( P<0.05), and it had moderate predictive accuracy for the stage Ⅱ of TTTS with an area under the curve of 0.766, a sensitivity of 64.71% and a specificity of 82.35%( P<0.001). Conclusions:More cerebroplacental blood flow discordances are observed and CPR disc is closely related to stageⅡof TTTS.

Construction of Dual-luciferase reporter gene vector of 3'UTR of the COL4A3 gene and validation of its targeting relationship with miR-299 / 中华显微外科杂志

Objective To construct a dual-luciferase reporter gene vector and validate the targeting relation ship between miR-299 and the COL4A3 gene,laying a foundation for the study on the effect of miR-299 in the chondrogenic differentiation of stem cells by regulating the COL4A3 gene.Methods This study was made from March,2018 to December,2018.Firstly,the potential binding sites between miR-299 and COL4A3-3'UTR were pre dicted using bioinformatics.Then,the wild and mutant COL4A3-3'UTR sequences were amplified by PCR and cloned into psiCHECK-2 plasmid to construct corresponding recombinant vectors.The vectors were validated by enzyme digestion and gene sequencing.Finally,the cells were resuscitated,amplified,transfected and divided into 4 groups:COL4A3-WT+miR-299/NC group,COL4A3-WT+miR-299-inhibitor/NC-inhibitor group,COL4A3-MUT+miR-299/NC group and COL4A3-MUT+miR-299-inhibitor/NC-inhibitor group.Each group contains 3 holes,respectively.Luciferase activity in each group was determined using a dual-luciferase assay kit.The statistical analysis was conducted and differences between groups were compared by t test.Probabilities lower than 5％(P＜0.05) were considered statistically significant.Results Enzyme digestion and DNA sequencing showed that the dual-luciferase reporter gene vector of psiCHECK-2-COL4A3 was constructed successfully.Luciferase assay demonstrated that in wild COL4A3 gene,luciferase activity reduced in the miR-299 transfection group (The average R/F value was 59.38％) compared with the NC group (The average R/F value was 100.00％),with a statistical significant difference (P＜0.05).In wild COL4A3 gene treated with inhibitor,luciferase activity increased in the miR-299-inhibitor group (The average R/F value was 153.98％) compared with the NC-inhibitor group (The average R/F value was 100.00％),with a statistical significant difference (P＜0.05).In mutant COL4A3 gene treated with inhibitor,no obvious statistical differences in luciferase activity were found between miR-299 transfection group (The average R/F value was 102.09％),miR-299-inhibitor group (The average R/F value was 108.51％) and NC group (The average R/F value was 104.70％),NC-inhibitor group (The average R/F value was 105.13％) and/9＞0.05.Conclusion The dual-luciferase reporter gene vector of the 3'UTR of the COL4A3 gene is constructed successfully.In addition,dual-luciferase assay further verifies the authenticity of miR-299 directly targeting the 3'UTR of the COL4A3 gene.

Investigation of relationship of serum and carcinoma levels of plasminogen activator inhibitor-1 with the invasion and metastasis of endometrial carcinoma / 中华老年医学杂志

Objective To investigate the relationship of serum and carcinoma levels of plasminogen activator inhibitor-1 (PAI-1)with clinical characteristics in patients with endometrial carcinoma.Methods Serum level of PAI-1 was determined by using enzyme-linked immunosorbent assay(ELISA) in patients with endometrial carcinoma (n =40),uterine prolapsed with normal endometrial tissues(n=40).The protein expression of PAI-1 in endometrial tissue was detected by using immunohistochemistry in patients with endometrial carcinoma and uterine prolapse patients.Results Serum level of PAI-1 was higher in patients with endometrial carcinoma than in uterine prolapse patients with normal endometrial tissues(19.43±7.12 μg/L vs.6.58±2.33 μg/L,P＜0.05).The rate of positive expression of PAI-1 was higher in endometrial carcinoma tissue than in uterine prolapse tissue[62.5 ％ (25/40) vs.7.5 ％ (3/40),P ＜ 0.01].Compared with early-stage endometrial carcinoma,advanced endometrial carcinoma had an increased rate of positive expression of PAI-1 (P ＜0.01).Compared with endometrioid adenocarcinoma,other pathological types of endometrial carcinoma had an increased rate of positive expression of PAI-1 (P ＜ 0.05).Poorly differentiated endometrial carcinoma versus highly differentiated endometrial carcinoma had an increased positive rate of PAI-1 (P ＜0.05).The rate of positive expression of PAI-1 was higher in endometrial carcinoma with myometrial invasion than without myometrial invasion(25/31 vs.0/9,P＜0.01).Conclusions The expression level of PAI-1 may be related to the invasion and metastasis of endometrial carcinoma.

Development of high-performance liquid chromatographic for quality and authenticity control of Chinese propolis.

A RP-high-performance liquid chromatography (HPLC) method was developed for quality control of Chinese propolis by simultaneous analysis of 12 flavonoids and 8 phenolic acids. The results showed that vanillic acid, rutin, myricetin, and luteolin were not detected in all of the analyzed propolis and poplar tree gum samples. The caffeic acid, ferulic acid and p-coumaric acid were not detected in poplar tree gum but were detected in propolis, which suggest that they are practical indexes of distinguishing propolis from poplar tree gum. The flavonoid profiles of poplar tree gum were found to be similar to those of propolis, which are dominated by pinobanksin, pinocembrin, 3-O-acetylpinobanksin, chrysin, and galangin. Therefore, the proposed method could be applied to exclude poplar tree gum from propolis with cafferic acid, ferulic acid, and p-coumaric acid as qualitative markers, and distinguish poplar source resin from other illegal substances, and evaluate the quality grading of poplar-type propolis with pinobanksin, pinocembrin, 3-O-acetylpinobanksin, chrysin, and galangin as qualitative and quantitative markers.

Cerebroplacental blood flow redistribution in monochorionic‐diamniotic twin pregnancies with selective fetal growth restriction / 中华超声影像学杂志

Objective To analyze the cerebroplacental blood flow distribution characteristics in monochorionic‐diamniotic ( MCDA ) twin pregnancies with selective fetal grow th restriction ( sFGR ) ,and investigate the relationship between co‐twin cerebroplacental blood flow discordances and co‐twin birth weight discordances ( BWdisc ) . Methods The cerebroplacental blood flow distribution characteristics and their discordances were analyzed retrospectively in 52 MCDA twin pregnancies with normal grow th ( control group) and 52 with sFGR ( case group) ,including the umbilical artery pulsatility index ( U A‐PI) ,middle cerebral artery peak systolic velocity ( MCA‐PSV ) ,middle cerebral artery pulsatility index ( MCA‐PI ) , cerebroplacental ratio ( CPR ) ,and their discordances ( U A‐PIdisc ,MCA‐PSVdisc ,MCA‐PIdisc and CPRdisc ) . Results Compared to the control group ,UA‐PIdisc ,MCA‐PIdisc and CPRdisc increased significantly ( all P ＜0 .01) . U A‐PIdisc ,MCA‐PIdisc and CPRdisc were related positively to BW disc as show n by correlation analyses ( r＝0 .488 ,0 .414 ,0 .592 ;all P ＜ 0 .001) ,and they had moderate predictive accuracy for sFGR with area under the curves of 0 .743 ,0 .662 and 0 .778 , with sensitivity of 48 .08% ,67 .31% and 71 .15% , and specificity of 92 .31% ,59 .62% and 78 .85% ( all P ＜ 0 .01 ) . M ultivariate Logistic regression analyses showed that only CPRdisc were independently associated with sFGR ( P ＜ 0 .05 ) . Conclusions More cerebroplacental blood flow discordances are observed in MCDA twin pregnancies with sFGR . Co‐twin cerebroplacental blood flow discordance is related to BW disc .