Abstract:Objective：To prepare and characterize magnetic albumin nanospheres conjugated with cetuximab（C225），which is anti-epithelial growth factor receptor（EGFR） monoclonal antibody，as well as their in vitro anti-lung cancer effect when combined with magnetic fluid hyperthermia. Methods：Fe3O4 nanoparticles（NPs）were prepared by chemical co-precipitation，and magnetic albumin nanospheres（MANs） were prepare by desolvation-crosslinking method，then C225 was further conjugated to synthesize C225-MANs using N-succinimidyl-3-（2-pyridyldithio）propionate（SPDP）. Their morphology，mean particle size，zeta potential，the antibody conjugation efficiency，iron content，specific cell-binding ability，and magnetothermal dynamic profiles were characterized. Lastly，the therapeutic effect of C225-MANs induced double targeted magnetic fluid hyperthermia on lung adenocarcinoma in vitro was evaluated using CCK-8 and flow cytometry（FCM）assay. Results：TEM micrographs showed that the prepared C225-MANs were approximately spherical. Fe3O4 NPs were well incorporated into the core of the nanospheres and the iron content was about 2.0 mg/mL. The mean hydrodynamic diameter was about 140 nm with negative charge. When placing in alternating magnetic field with the output frequency of 200 kHz and the output current of 20 A，the corresponding C225-MANs with different Fe concentrations could rise to a steady temperature ranging from 39.3 ℃ to 52.0 ℃，and the temperature could keep stable without any change after 30 min heating. The fluorescence percentage detected by FCM analysis revealed that the antibody conjugation efficiency was 79.74%. The ability of nanospheres targeting GLC-82 cells in vitro was confirmed by immunofluorescence experiments and magnetic resonance imaging. The proliferation rate of double C225 and magnetic-targeted combined group was significantly lower than any other therapy group（P＜0.05），while the apoptotic rate of the double C225 and magnetic-targeted combined treatment group were significantly higher than any other therapy group（P < 0.05）. Conclusion：C225-MANs had good targeting effect on GLC-82 cells at cellular level. Moreover，dual-targeted magnetic fluid hyperthermia combined with molecular targeted therapy can effectively combat lung cancer in vitro.

Abstract:Objective：To prepare a monoclonal antibody against SraPL-lectin，a serine-rich repeat protein of Staphylococcus aureus，and analyze its function. Methods：The SraPL-lectin gene was amplified by PCR from USA 300 DNA，and inserted into pET28a plasmid. The pET28a- SraPL-lectin was transferred into E.coli. BL21（DE3）and induced with 0.1 mmol/L isopropyl-β-d-thiogalactoside（IPTG）for 6 h at 25 ℃. The recombinant protein was purified by His label affinity chromatography. The purified protein was used as an antigen to generate an anti-SraPL-lectin monoclonal antibody. The mAb was identified by ELISA and Western blot. A549 cells were pre-incubated monoclonal antibodies with different final concentrations. Mice were injected with 1 μg anti-SraPL-lectin monoclonal antibodies one day before challenge. The effect of adhesion and invasion was counted by plating bacteria on TSA. Results：We successfully constructed a prokaryotic expression pET28a-SraPL-lectin vector，and the SraPL-lectin recombinant protein was expressed and purified. We also generated a clone of hybridoma with SraPL-lectin binding activity，and obtained anti-SraPL-lectin monoclonal antibody purified from mouse ascites by protein G column. The titer of mAb was 1∶320 000. Pre-incubation of A549 cells with a final concentration of 100 ng/mL for 2 h significantly reduced the adherence and invasion of S. aureus on A549 cells. The number of bacteria in blood was significantly decreased when mice were administered 1 μg anti-SraPL-Lectin monoclonal antibodies. Conclusion：The anti-SraPL-Lectin monoclonal antibody prepared in this study significantly reduced the adherence and invasion of S. aureus on host cells. This study lays the foundation for the development of SraPL-Lectin as a target for the prevention and treatment of S. aureus infection in future.

Abstract:Objective：To detect the expression of specificity protein 1（SP1）in the renal tissues of rats with Thy-1 nephritis（Thy-1N）and in the glomerular mesangial cells（GMCs）stimulated with sublytic C5b-9，and investigate the role of SP1 expression in sublytic C5b-9-induced GMC proliferation. Methods：Rat Thy-1N was induced，and the cultured rat GMCs were stimulated with sublytic C5b-9. The expression of SP1 in the renal tissues of rats with Thy-1N（in vivo）and in the GMCs stimulated with sublytic C5b-9（in vivo）was detected by real-time PCR and Western blot，respectively. Meantime，SP1 expression plasmid（pEGFP-N1/SP1-His）and SP1 shRNA（shSP1）were constructed. Above-mentioned plasmids and control plasmids were transfected into rat GMCs followed by sublytic C5b-9 stimulation or not. The expression levels of SP1 were evaluated by real-time PCR and Western blot，and the cellular proliferation was determined by CCK-8. Results：The expression levels of SP1 in the renal tissues of rats with Thy-1N and in the GMCs stimulated with sublytic C5b-9 were markedly increased with similar time points. SP1 overexpression could markedly promote GMCs proliferation，while SP1 knockdown could obviously reduce sublytic C5b-9-induced GMCs proliferation. Conclusion：Sublytic C5b-9 promotes rat GMCs proliferation through up-regulation of SP1.

Abstract:Objective：To observe the protective effect and mechanism of liguzinediol on heart function in acute heart failure rats induced by propafenone. Methods：The rat model of acute heart failure was replicated by propafenone intravenous injection，and different doses of liguzinediol（5.0，10.0 and 20.0 mg/kg）and the positive drug of digilanid C（0.045 mg/kg）were given. The left ventricular pressure was recorded by the RM6240 multichannel physiological signal acquisition and processing system for 0，5，10，20，40，60，90 and 120 min. The changes of maximum ascending/ descending rate（±dp/dtmax），left ventricular systolic pressure（LVSP），mean systolic pressure（MSP），mean diastolic pressure（MDP）and heart rate（HR）were observed，and calcium transport related protein expression was used to detect the mechanism of liguzinediol by Western blot. Results：Liguzinediol 5.0，10.0 and 20.0 mg/kg could effectively increase the +dp/dtmax，-dp/dtmax，LVSP，MSP，MDP and HR in model rats. Western blot and q-PCR results showed that the expression of Ca2+/calmodulin-dependent protein kinaseⅡ（CAMKⅡ），P-CAMKⅡ，sarcoplasmic reticulum Ca2+-ATPase2a（SERCA2a）and P-phospholamban（P-PLN）increased significantly after intervention with liguzinediol. There was no significant difference in ryanodine receptor 2（RyR2） and FK506 binding protein 12.6（FKBP12.6） expression. Conclusion：Liguzinediol can increase the intracellular calcium concentration and improve heart failure，and the mechanism may be through the regulation of CAMKⅡ.

Abstract:Objective：The goal of this study is to explore the effects of ulinastatin on acute lung injury in fetal rat induced by fetal distress，and provide support for developing a new treatment of respiratory distress syndrome. Methods：A total of 16 SD pregnant rats were randomly divided into four groups：the control group（group C），the ulinastatin control group（group S），the fetal distress group（group A），and the ulinastatin treatment group（group U）. Group C and group S were sham operation groups. Fetal rat intrauterine distress model was set up in both group A and group U. Thirty minutes before the operations，the pregnant rats in group S and group U received injection of ulinastatin 50 000 U/kg through femoral vein. Five living fetal rats were removed in each pregnant rat. The blood gas analysis and the lung wet / dry weight ratio（W/D）of fetal rats were determined. The pathological changes in the lung tissue of fetal rats were observed by H＆E staining，and the protein expression of NF-κB p65 were measured by Western blot. Furthermore，the levels of TNF-α and IL-6 in the fetal lung were measured by ELISA. Results：All the results in group S had no significant difference with group C（P >0.05）. The pH value and partial pressure of oxygen were significantly lower in group A than group C，and those in group U were obviously higher than group A（P＜0.05）. The partial pressure of carbon dioxide，lactic acid content，the lung W/D，and the protein expression of NF-κB p65，the levels of TNF-α and IL-6 of fetal rats were all significantly increased in group A than group C（P＜0.05），and those in group U were obviously lower than those in group A（P＜0.05）. The congestion and edema of the lung tissue in group U were obviously alleviated than that of group A. Conclusion：Ulinastatin can alleviate the degree of acute lung injury in fetal rat induced by fetal distress by decreasing the protein expression of NF-κB p65，the levels of TNF-α and IL-6，inhibiting the inflammatory response and increasing the partial pressure of oxygen.

Abstract:Objective：To investigate the susceptibility of four genetic variations of prostate cancer non-coding RNA 1（LncRNA PRNCR1）to the risk and pathological characteristics of gastric cancer. Methods：A total of 300 gastric cancer cases and 300 health controls were enrolled in this case-control study. The Mass-array gene analysis platform was applied to genotype the enrolled participants. Enzyme-linked immunosorbent assay was used to detect the H. pylori infection. Results：There were significant differences of LncRNA PRNCR1 rs16901946 genotypes distribution between the cases and the control group（P=0.018），and the AG（ORadjusted=1.41，95%CI：1.00-1.98，P=0.048），GG（ORadjusted=2.57，95%CI：1.15-5.75，P=0.022）and AG/GG genotype（ORadjusted=1.51，95%CI：1.09-2.09，P=0.014）were associated with increased risk of gastric cancer，respectively. The results of subgroup analysis showed that LncRNA PRNCR1 rs16901946 was associated with the risk of gastric cancer in the subgroup of male（AG/GG vs. AA：ORadjusted=1.82，95% CI：1.23-2.69，P=0.003），and individuals with H. pylori infection（AG/GG vs. AA：ORadjusted=1.83，95%CI：1.16-2.89，P=0.009）. In addition，LncRNA PRNCR1 rs16901946 was associated with risk of cardia cancer（AG/GG vs. AA：ORadjusted=1.58，95% CI：1.10-2.28，P=0.011）. Similarly it was associated with risk of gastric cancer in clinical stage T1-T2（AG/GG vs. AA：ORadjusted=1.88，95%CI：1.18-2.98，P=0.008）but not in T3-T4. Conclusion：This study indicates that LncRNA PRNCR1 rs16901946 is associated with the increased risk of gastric cancer，especially for male and individuals with H. pylori infection.

Abstract:Objective：To investigate the effect of Pol ι expression on overall survival of esophageal squamous-cell carcinom（ESCC） patients with postoperative adjuvant radiotherapy. Methods：The correlation between Pol ι expression and overall survival of 97 ESCC patients with postoperative adjuvant radiotherapy was analyzed using immunohistochemical method. Results：There were 20 cases with low expression of Pol ι and 77 cases with high expression of Pol ι in these patients. Survival curve showed Pol ι expression was negatively correlated with overall survival of ESCC patients with postoperative adjuvant radiotherapy，and COX regression analysis indicated that Pol ι expression was one of the independent prognostic factors of postoperative adjuvant radiotherapy for esophageal cancer. Conclusion：Pol ι may be a potential molecular markers for postoperative adjuvant radiotherapy in esophageal cancer.

Abstract:Objective：To evaluate the expression of microRNA-145（miR-145）in epithelial ovarian cancer tissue and serum，and investigate the diagnosis and prognosis value of miR-145 as a biomarker for epithelial ovarian cancer. Methods：The expression of miR-145 in serum from 45 cases of epithelial ovarian cancer and 45 cases of healthy control，as well as in ovarian tissues from 45 cases of epithelial ovarian cancer and 8 cases of healthy control，was detected by real-time PCR. Results：The expressions of miR-145 in the serum and ovarian tissues of patients with epithelial ovarian cancer were significantly lower than those in healthy control（P＜0.01）. The expression of miR-145 decreased with the progression of the clinical stages（P＜0.05），but it is not correlated with age，histological classification，lymph node metastasis，histological grading and the increasing of CA125（P＞0.05）. ROC curves revealed that serum miR-145 might be a promising diagnostic biomarker for patients with epithelial ovarian cancer. Conclusion：MiR-145 act as a tumor suppressor in the development of epithelial ovarian cancer. MiR-145 may serve as a biomarker for diagnosis and predictor of epithelial ovarian cancer.

Abstract:Objective：To evaluate the short and medium-clinical effect of acupuncture therapy in treating hot flashes in patients with breast cancer by meta-analysis. Methods: This systematic research analyzed all randomized controlled trials （RCTs） relating with the effects of acupuncture treatment for hot flashes of breast cancer patients in the CNKI，CBM，VIP，Wanfang and PubMed （30th June 2017 ago）. The quality of RCTs was assessed by Cochrane systematic review. Multiple overall effects of results were analyzed with meta-analysis，and evaluated systematically by RevMan5.3 software. The heterogeneity was evaluated. Results: In the follow-up visit of 3 months （SMD=-0.36，95%CI: -0.61~-0.10） and 6 months （SMD=-0.41，95%CI: -0.64~-0.18），acupuncture improved the syndrome systematically （P=0.006，P=0.000 5，respectively）. Conclusion: The results of clinical researches with high study quality show that acupuncture treatment can significantly improve the syndrome of hot flashes in short- and medium-term in breast cancer patient.

Abstract:Objective：To analyze risk factors of hyperuricemia（HUA）in patients with newly diagnosed type 2 diabetes mellitus（T2DM）. Methods：A total of 735 patients with newly-diagnosed T2DM from the First Affiliated Hospital of NMU were enrolled in this case-control study from March 2010 to March 2017. The general clinical data and biochemical indicators of the patients were collected，and the related factors affecting the level of serum uric acid were analyzed. Results：The patients were divided into two groups according to with or without HUA，110 patients with HUA while 625 patients without HUA. The age，lipoprotein A，and estimated glomerular filtration rate（eGFR）in the T2DM with HUA group were all significantly lower than those patients without HUA（P＜0.05）. Besides，the body mass index（BMI），alanine aminotransferase（ALT），aspartic transaminase（AST），gamma-glutamyl transpeptidase（γGGT），triglyceride（TG），creatinine（Cr）and retinol binding protein（RBP）of the T2DM with HUA group were significantly higher than those of T2DM without HUA group（P＜0.05）. Logistic regression analysis revealed that male，low age，high BMI，high TG and low eGFR in the newly diagnosed T2DM patients were independent risk factors complicated with HUA. Conclusion：In the newly diagnosed T2DM patients，males who were relatively young，overweight，with hypertriglyceridemia and low eGFR，should be paid more attention on the comprehensive management of hyperuricemia.

Abstract:Objective：To investigate the clinical value of whole-genome chromosomal microarray analysis（CMA）in the genetic diagnosis of chorionic villi and stillbirths and the relationship between spontaneous miscarriage and chromosomal aberrations，and to provide genetic counseling for couples with spontaneous abortion. Methods：A total of 91 abortion samples were tested by Affymetrix Cytoscan 750 K and analyzed with Chromosome Analysis Suite（ChAS）software. The detected copy number variations（CNVs）were aligned with public literatures and known CNVs listed in databases，such as DGV，DECIPHER，OMIM，ISCA，UCSC，and ClinVar. Results：All 91 cases（100%）were successfully detected，among which 61（67.03%）were chromosomal abnormalities. Forty（65.57%）of the 61 specimens were aneuploidy（30 trisomy and 10 monosomy），10 specimens（16.39%）were structural anomalies，2 specimens（3.28%）were polyploid，5 specimens（8.20%）were mosaicisms，and 4 specimens（6.56%）showed loss of heterozygosity or uniparental disomy. The structural anomalies were identified in 10 patients，which involved microdeletions and microduplications on 1p36.33p31.1，4p16.3，7q36.2q36.3，22q11.21，and four cases with small segmental deletion and duplication at the terminal end of the long and short arm of two chromosomes，which highly indicated that one of their parents could be carriers of submicroscopic reciprocal translocation. Conclusion：Whole-genome chromosomal microarray analysis could be used as a first-line method for genetic diagnosis of the miscarriages，due to its high-throughput，high resolution，high-accuracy，rapid-analysis and without cell culture. The identification of the cytogenetic cause of spontaneous miscarriage can facilitate estimation of recurrence risks for future pregnancies. It can be used as the main detection technique for genetic diagnosis of abortions.

Abstract:Objective：To establish a method for the determination of elements in serum of patients with bloodstream infection by inductively coupled plasma mass spectrometry（ICP-MS）. Combined with metallomics，we obtained the metal element profile of E. coli bloodstream infection（BSI）and screened key element markers，which provides proof for diagnosis and prevention of bloodstream infection. Methods：Taking 6Li，89Y，45Sc，115In，159Tb，and 209Bi as internal standards，ICP-MS was used to measure 68 kinds of metal elements in serum of 199 normal humans，diluting serum samples and internal standard method for correction，with kinetic energy discrimination to eliminate mass spectroscopic interference. Meanwhile，68 kinds of metal elements in serum from 19 BSI patients with E. coli infection and 19 healthy people were tested by ICP-MS. Statistical analysis was performed using principal components analysis（PCA）and partial least squares discriminate analysis（PLS-DA），and the differences were analyzed. Results：Based on ICP-MS，the profile of metal elements in blood serum of patients with BSI and healthy people was established，and the differential elements Zn，Ce，Pd，Cu，Sn，Se，Ba，Au and Fe in the serum of the patients with BSI and healthy controls were screened out. Conclusion：We established multi-elementary metal profiles using ICP-MS in human serum，which is acceptable for a clinical routine analysis. There were differences in the serum elements between BSI patients and the healthy controls. The results provide a basis for the diagnosis，prevention and treatment of blood bacterial infections from the perspective of metallomics.

Abstract:Objective：To assess whether the first blood lipid test in outpatients is necessary to detect low-density lipoprotein cholesterol（LDLC） in order to achieve grading test of medical examination，reduce the labor of medical inspectors and reduce medical expenses. Methods：Total cholesterol（CHO），triglycerides（TG），high-density lipoprotein cholesterol（HDLC），and LDLC data were obtained from Laboratory Information System（LIS） based on outpatients in 4 years（2013-2017）. CHO，HDLC and LDLC were measured using TBA2000FR biochemical analyzer. nonHDLC was calculated with CHO minus HDLC. Correlation between CHO，nonHDLC and LDLC were analyzed using Spearman’s rank approach. Receiver operating characteristics（ROC） curve analysis was used to evaluate the predictive of CHO and nonHDLC for abnormal LDLC. Results：Both CHO（r=0.843）and nonHDLC（r=0.862）were significantly positively correlated with LDLC. Area under curve of CHO and nonHDLC for predicting abnormal LDLC（＞3.40 mmol/L and＞4.10 mmol/L）were 0.941 and 0.948，0.967 and 0.970，respectively（P＜0.001）. Optimal thresholds of prediction abnormal LDLC were 3.85 mmol/L and 4.33 mmol/L for nonHDLC. Based on these optimal thresholds，the sensitivity of nonHDLC to predict LDLC positive results was 91.1% and 97.6%，and the specificity was 85.4% and 87.3%，respectively. Less than 3.33% and 0.16% of tests with abnormal LDLC might be missed，but approximately 66.3% and 80.6% of the LDLC tests could be eliminated. Conclusion：We recommend that LDLC measurement is not necessary for the first test of blood lipids in outpatients. About 65% of LDLC tests would be reduced. Moreover，nonHDLC is a better index than CHO to predict abnormal LDLC.