Objective : To explore the role of lysophosphatidylcholine acyltransferase 3(LPCAT3) in Erastin-induced ferroptosis of acute myeloid leukemia(AML) cells and its related molecular regulatory mechanisms. Methods : Tetrazolium salt(MTS) method was used to detect the sensitivity of different AML cells to the classic ferroptosis inducer Erastin, real time quantitative polymerase chain reaction(qPCR) was used to detect the basal expression level ofLPCAT3mRNA, and the correlation between them was analyzed. Lentivirus-mediatedLPCAT3overexpression AML cell lines(OE group) and negative control lines(NC group) were constructed. After Erastin intervention, MTS, flow cytometry, and micromethods were used to detect cell viability, lipid reactive oxygen species(ROS), and Malondialdehyde(MDA), respectively. qPCR and Western blot were used to detect unfolded protein response(UPR) classic pathway signaling molecules(PERK, ATF4, GRP78, etc.) expression levels. The above ferroptosis-related indicators were detected after combined intervention with the UPR inhibitor 4-phenylbutyric acid(4-PBA), and the regulatory relationship was analyzed. Results : Four different types of AML cells had different sensitivities to ferroptosis, among which K562 cells were relatively insensitive. The IC50of the four types of AML cells to Erastin was negatively correlated with the expression level ofLPCAT3(r=-0.919,P<0.001). After Erastin intervention, the cell viability of K562 cells in the OE group was significantly inhibited by Erastin compared with the NC group(P<0.001), and the levels of lipid ROS and MDA increased(P<0.001). The results of qPCR and Western blot showed that, compared with the NC group, the mRNA and protein expression of UPR classic pathway moleculesPERK,ATF4, andGRP78mRNA and protein increased in the OE group(P<0.01). After inhibiting the UPR pathway by 4-PBA, the viability of K562 cells decreased(P<0.01), and lipid ROS and MDA levels increased(P<0.01) compared with the uninhibited state. Conclusion Overexpression ofLPCAT3can promote ferroptosis in K562 cells, and this process is negatively regulated by the classical UPR pathway PERK/ATF.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Objective:To investigate the clinical characteristics and the risk of major adverse cardiac events within 1 year of middle-aged and elderly in-patients with acute decompensated and mid-range ejection fraction heart failure(HF)in the medical alliance setting.Methods:A retrospective cohort study was conducted among a total of 180 in-patients with acute decompensated heart failure in Cardiovascular Hexi Hospital Consulting Area of Tianjin Chest Hospital.According to ejection fraction measured by echocardiogram, the in-patients were classified into three groups: heart failure with reduced ejection fraction(HFrEF)group(n=70, 38.9%), HFmEF group(n=50, 27.8%), and heart failure with preserved ejection fraction(HFpEF)group(n=60, 33.3%). Clinical feature and 1-year prognosis between different groups were compared.Results:Univariate Cox regression analysis of 1-year all-cause death and cardiovascular death showed that there was no significant difference between HFrEF group and HFmEF group, HFpEF group and HFmEF group(all P>0.05); 1-year readmission analysis of heart failure showed that 47.1%(33 cases)of HFrEF group was higher than 24.0%(12 cases)of HFmEF group, 48.3%(29 cases)of HFpEF group was higher than HFmEF group( HR=2.307, 2.368, 95% CI: 0.187-4.480, 1.207-4.644, respectively, all P<0.05); The major 1-year cardiovascular events were 57.1%(40 cases)higher in the HFrEF group than 34.0%(17 cases)in the HFmEF group( HR=2.053, 95% CI: 0.187-4.408, P< 0.05). Multivariate analysis showed that the 1-year risk of major cardiovascular events was significantly different between HFmEF group and HFpEF group( HR=0.477, 95% CI: 0.241-0.941, P< 0.05). Pulmonary heart disease( P< 0.05), atrial flutter and/or atrial fibrillation( P< 0.01), New York Cardiology class Ⅳ( P< 0.01)were risk factors for death.Hypertension and cor pulmonale were the risk factors for readmission in patients with heart failure(all P< 0.01). Conclusions:The clinical characteristics of inpatients with HFmEF in the medical alliance setting tended to be consistent with those with HFrEF, while the feature of ischemic heart disease was more prominent in HFmEF.The 1-year risk of heart failure readmission in HFmEF group was significantly lower than that in HFpEF and HFrEF group, and the risk of all-cause mortality and cardiovascular mortality at 1 year was not significantly different among the three groups.

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Objective To study the diagnostic value of narrow-band imaging ( NBI) combined with endoscopic ultrasonography ( EUS) for ampullary tumors. Methods A total of 21 patients suspected with ampullary lesions by imaging or endoscopic examination from December 2015 to March 2017 were enrolled in this prospective study. All patients underwent NBI and EUS, and 20 patients underwent biopsy. The type of ampullary tumor was predicted by preoperative examination, and appropriate treatment methods were chosen. The final diagnosis was confirmed by biopsy, surgical pathology, and clinical follow-up for more than 6 months. The accuracy of NBI combined with EUS and biopsy in diagnosis of ampullary malignant tumors was calculated according to the gold standard. The Chi-square test was used to compare diagnostic accuracies. Results The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of NBI combined with EUS in diagnosis of ampullary malignancies were 94. 1％ (16/17), 100. 0％ (4/4), 95. 2％ (20/21), 100. 0％ (16/16), and 80. 0％ (4/5), respectively. The corresponding indicators of preoperative biopsy were 41. 2％ ( 7/17) , 100. 0％ ( 3/3) , 50. 0％ ( 10/20) , 100. 0％ ( 7/7) , and 23. 1％( 3/13) , respectively. The accuracy of NBI combined with EUS in diagnosing ampullary malignant tumor was significantly higher compared with preoperative biopsy ( P=0. 004) . Conclusion NBI combined with EUS can more accurately predict benign or malignant ampullary tumor, and better guide the choice of surgical methods compared with preoperative biopsy.

Objective To study port-externalized tissue expansion technique and to provide a safe,simple and effective skin tissue expansion for extensive skin lesion reconstruction.Methods A lot of 247 patients with extensive skin lesion were chosen for the study.400-1000 ml tissue expanders were used for overexpansion and repeated expansion.Expanders were inserted into subcutaneous pockets through mini-invasive incisions and intermittent injection was carried out 4 days after first stage operation.After expansion finished,expanded skin flaps were transplanted for skin lesion reconstruction.During inflation period,complications and expander failures were observed.Results In 741 expanders of 247 patients,complication rate was 16.5 ％ with total 122 cases of complications.Expansion success rate was 98.9 ％ with 8 expander failures.Expanders were inflated successfully between 64-113 days,with average 92.7 days.The inflated saline volume was 2.38 times of marked expander capacity.In this group of 733 expanded skin flaps,721 flaps were well survived with 98.4％ success rate.276 expanded flaps of 116 patients were followed up for one to five years post-operation.The width of incision scar was between 0.3-1.2 mm.Conclusions The port-externalized overexpansion with giant expanders is a safe,simple and feasible technique,which is worthy of further clinical application.

Objective To evaluate measurement of the submucosal thickness with endoscopic ultrasonography (EUS) for activity of Crohn disease (CD).Methods Ten patients with active stage of CD and 10 healthy controls (HC) underwent EUS.Simple endoscopic score for Crohn disease(SES-CD)and submucosal thickness at the most severe lesions were measured and recorded.Submucosal thickness of the same region in CD patients were measured at remissive stage.In order to analyze the relationship between submucosal thickness and the stage of CD, submucosal thickness were compared among patients at active stage of CD, remissive stage of CD and HC.And the cut-off value of submucosal thickness was calculated to diagnose the stage of CD.Results The mean submucosal thicknesses of active stage and remissive stage of CD were 6.48±1.95 mm and 2.47±1.08 mm,respectively (P<0.01).The correlation analysis showed that submucosal thickness had a positive correlation with Crohn disease activity index(CDAI)(r=0.708,P<0.01) and SES-CD(r=0.807,P<0.01).Receiver operating characteristic curve analysis was used for 10 cases of CD patients and the area under the curve was 0.985(P<0.01).The cut-off value of submucosal thickness to diagnose active stage of CD was 3.85 mm, and the sensitivity and specificity reached 100% and 90% respectively.The Youden index was 0.9.Conclusion Measurement of gastrointestinal submucosal thickness by EUS could contribute to evaluate the stage of CD and to guide clinical treatment.

Objective To discuss application results in early swelling in Pilon fracture by traditional Chinese medicine hot and cold spray with dorsal foot acupoint massage. Methods The 120 clinical datas of patients with Pilon fracture bone surgery in our hospital were analyzed,and divided into the control group and the observation group with 60 coses in each group. The elimination of time in patients with swelling and muscle strength rating of two groups patients with pilon fracture were detected,nursing after 24 h,120 h, the swelling curative effect of two groups patients with pilon frac-ture patients were detected. Results The elimination of time in patients with swelling and muscle strength rating of the observation group patients with pilon fracture were better than control group (t=7.75,χ2=5.13, 3.62, 3.13, 4.95, 24.56), nursing after 24 h, 120 h, the swelling curative effect of the observation patients with Pilon fracture were better than control group, the difference were statistical significance (χ2=14.25,30.81,P<0.05). Conclusion The application results in early swelling in Pilon fracture by traditional Chinese medicine hot and cold spray with dorsal foot acupoint massage, which can obviously eliminate swelling and improve muscle strength,the curative effect is very good,and worth to be used.

The non-human primates have been profoundly being used to study the human reproductive models .It is the prerequisites to set up the operating rooms including hardware and software on reproductive experiments and transgenic manipulation.Thus, Combined with the practice of our center , both the construction and the associated problems about the IVF-ET operating rooms were discussed in this paper .It will provide some help for the related researches .