Objective To assess the diagnostic value of Logistic model in differentiating between malignant and benign ovarian lesions. Methods Thirty-five indexes of clinical and ultrasound data were recorded in 601 ovarian lesions confirmed by surgical pathology. The Logistic model was developed on a training set( n - 400) and tested on a test set( n = 201). Results Variable selection resulted in a set of 10 variables for the models: personal history of ovarian cancer, maximal diameter of the lesion, maximal diameter of the solid component, multilocular-solid tumor, solid tumor, ascites, flow within papillation, irregular walls, very strong intratumoral blood flow (i. e. color score 4) and acoustic shadows. Test set area under the receiver-operating characteristics curve was 0.963 with a sensitivity 93.9% and a specificity 93. 1 %. Conclusions Logistic model can accurately separate malignant from benign ovarian masses.

Background Study determined that retinitis pigmentosa has a similar pathogenesis mechanism to Alzheimer disease,and activity of cyclin-dependent kinase 5 (Cdk5) and its activators participates in the degeneration of central nervous system.Roscovitine,an inhibitor of Cdk5,can suppress activity of Cdk5/p25 pathway and therefore inhibit cell apoptosis.However,the influence of roscovitine on retinitis pigmentosa(RP) is unclear.Objective This study was to investigate the expressions of p35,p25 and tau in the retinas of RCS rats.Methods Roscovitine of 4 μl was intravitreally injected in the right eyes of 12 SPF 17-day-old RCS rats,and the fellow eyes were not intervened as the control eyes.The rats were sacrificed on eighth day (postnatal 25 days) and eighteenth day (postnatal 35 days),and whole retinas were isolated to evaluate the relative expressions of Cdk5,p35,p25 and tau phosphorylation by Western blot,and the activity of Cdk5/p25 was analyzed by quantitative colorimetric assay.The results were compared between the right eyes and fellow eyes by paired t test.The use and care of the rats complied with Ethic Statement of Experimental Animal of Ningxia Medical University.Results In the eighth and eighteenth day after injection,the relative expression values (A values) of p35 in rat retinas were 1.186±0.019 and 1.069± 0.019 in the injected eyes,showing significant decreases in comparison with 1.364±0.016 and 1.214±0.008 of the fellow eyes (t =-6.294,-6.477,both at P＜0.05);the relative expression values (A values) of p25 in rat retinas were 0.312±0.009 and 0.269±0.018 in the injected eyes,which was significantly lower than 0.595±0.013 and 0.473±0.011 of the fellow eyes (t=-36.508,-11.879,both at P＜0.05).No significant difference was found in the relative expression of Cdk5 protein between the injected eyes and the fellow eyes in various time points after injection (both at P＞0.05).The activities of Cdk5/p25 were (0.003 83 ±0.000 14) mol/(s · mg) and (0.002 01 ± 0.000 11) mol/(s · mg) in the injected eyes,with significant decreases in comparison with the (0.005 47±0.000 27)mol/(s · mg)and (0.003 35±0.000 15) mol/(s · mg) of the fellow eyes (t=-9.152,P=0.000;t=-9.248,P=0.000),and the tau phosphorylation levels followed the same pattern in the eighth and eighteenth day after injection (t =-9.854,-6.744,both at P＜0.05).Conclusions Intravitreal injection of roscovitine can inhibit the activity of Cdk5/p25 and tau phosphorylation level in retinas of RCS rats to certain extend.

Objective To explore the influence of icotinib in the apoptosis of the human salivary adenoid cystic carcinoma cells ACC-M, and to clarify the mechanism of icotinib for the treatment of salivary adenoid cystic carcinoma.Methods The ACC-M cells were randomly divided into control group,2,4,8μmo1·L-1 icotinib groups,p38-MAPK inhibitor SB203580 (20μmol· L-1 )group,SB203580 (20 μmol· L-1 )+4μmo1 · L-1 icotinib group;the cells were collected 4 h after treatment.The viability of ACC-M cells was measured by MTT assay.The apoptosis of ACC-M cells was assessed by caspase-3 activity kit. The expression of p-p38-MAPK protein was determined by Western blotting analysis.Results Compared with control group,the inhibitory rates of growth of the ACC-M cells in icotinib groups were significantly decreased (P<0.05 ), and the activities of caspase-3 were increased (P<0.05),and the expression levels of p-p38-MAPK were significantly increased (P<0.05).Compared with 4μmo1·L-1 icotinib group,the expression level of p-p38-MAPK in SB203580+icotinib group were decreased (P < 0.05 ), and the activity of caspase-3 was decreased dramatically (P < 0.05 ). Conclusion Icotinib may induce the apoptosis of ACC-M cells through the activation of p38-MAPK signaling pathway.

BACKGROUND: Soybean isoflavone has a variety of bioactivities and its antioxidation becomes a hot spot of research in recent years. At present,the research of soybean isoflavone places more emphasis on animal experiment and clinical observation,but lacks research on cellular level of human body.OBJECTIVE: To observe the influence of soybean isoflavone in vascular endothelial cells with oxidative damage.DESIGN: Controlled trial and observation.SETTING: Central Laboratory, Institute of Public Health,Tianjin Medical University.MATERIALS: The experiment was completed in the Central Laboratory,Institute of Public Health,Tianjin Medical University from January to July 2002.The experimental materials included vascular endothelial cell strain in human umbilical vein,low density lipoprotein,soybean isoflavone and vitamin E,etc.METHODS: The vascular endothelial cells were cultured in vitro.The experiment was divided into 6 groups: blank control group,oxidative damage control group (malondialdehyde content was 1 μmol/L),oxidative damage+vitamin E control group(vitamin E was 50 μmol/L) and oxidative damage+soybean isoflavone 10,50,100 μ mol/L control group. The endothelial cells,which were joined with vitamin E and soybean isoflavone of different concentrations in advance to be incubated for 24 hours,were affected by oxidized low density lipoproteins and then cultured continually for 24 hours.All the indexes of antioxidation were determined in both extra-cell and intra-cell.MAIN OUTCOME MEASURES: Malondialdehyde content,activity of superoxide dismutase and glutathione peroxidase,the release condition of lactate dehydrogenase and productive quantity of nitrogen monoxide inside and outside the endothelial cells of each group.RESULTS: ①Comparison of malondialdehyde content,the activity of superoxide dismutase and glutathione peroxidase in endothelial cells of each group: The malondialdehyde content was higher significantly in oxidative damage control group than in blank control group (P ＜ 0.01),but the activity of superoxide dismutase and glutathione peroxidase was lower significantly in oxidative damage control group than in blank control group(P ＜ 0.01).The malondialdehyde content was lower significantly in oxidative damage+vitamin E control group,oxidative damage+soybean isoflavone 10,50,100 μmol/L control group than in oxidative damage control group(P ＜ 0.01),but the activity of superoxide dismutase and glutathione peroxidase was higher significantly in oxidative damage+vitamin E control group,oxidative damage+soybean isoflavone 10,50,100 μ mol/L control group than in oxidative damage control group (P ＜ 0.01). ②Comparison of the release condition of lactate dehydrogenase and the productive quantity of nitrogen monoxide in endothelial cells of each group: The release percentage of lactate dehydrogenase was higher significantly in oxidative damage control group than in blank control group (P ＜ 0.01),but the productive quantity of nitrogen monoxide was lower significantly in oxidative damage control group than in blank control group(P ＜ 0.01).The release percentage of lactate dehydrogenase was lower significantly in oxidative damage+vitamin E control group,oxidative damage+soybean isoflavone 10,50,100 μmol/L control group than in oxidative damage control group (P ＜ 0.01),but the productive quantity of nitrogen monoxide was higher significantly in oxidative damage+vitamin E control group,oxidative damage+soybean isoflavone 10,50,100 μmol/L control group than in oxidative damage control group(P ＜ 0.01).CONCLUSION: Soybean isoflavone can alleviate the oxidative damage in vascular endothelial cells,caused by oxidized low density lipoprotein,possibly through such antioxidization indexes as malondialdehyde content,the activity of superoxide dismutase and glutathione peroxidase,the release condition of lactate dehydrogenase and the productive quantity of nitrogen monoxide,etc.

Objective To study the value of multi-slice spiral CT(MSCT)and high-resolution MR in evaluating the feasibility of total mesorectal excision(TME)for rectal cancer before operation.Methods Sixty-three cases biopsy-proven rectal cancer underwent MSCT and high-resolution MR examination,including CT in 35 and MR in 28.The involvement of mesorectal fascia,lymph nodes extra-fascia and the curve distance between the lesions and the anal margin were observed and compared with that of operation and pathology.Results In evaluating the involvement of mesorectal fascia,two patients were oversteped by CT and one patient by MRI,and there was no significantly statistic difference between CT and MRI.The curve distance between lesions and anal margis could be measured accurately on sagittal section of MR images in all cases.Sixteen patients had been found having metastatic lymph nodes at extra-mesorectal fascia,twelve of them were biopsy-proven.Thirty-seven patients had been found having metastatic lymph nodes at intra-mesorectal fascia,but 4 of them were inflammatory biopsy-proven.Conclusion MSCT and high-resolution MR are of significant values in evaluating the feasibility of TME in the patients with rectal carcinoma.

Sef (similar expression to fgf genes) was identified as a feedback antagonist of FGF signaling in zerbrafish, mouse and human. Sefhas been reported to function in different ways, however the regulation of Sef stability remains unknown. The possible role of c-Cbl in the regulation of Sef protein degradation was investigated. Results from coimmunoprecipitation and immunostaining assays reveal that hSef colocalizes and interacts with c-Cbl. Data suggest that the interaction between hSef and c-Cbl results in the ubiquitination and subsequent degradation of the hSef protein. It was proposed that c-Cbl may serve as a modulator to regulate Sef protein stability during FGF signal transduction.

OBJECTIVETo study the HBsAg transient expression in HepG2 or COS-7 cells with eukaryotic expression plasmids inserting HBsAg gene (pCI-S and pcDNA3.1-S) and the efficacy of naked DNA immunization in mice.

METHODSFirstly, the recombinant plasmids of pCI-S and pcDNA3.1-S were constructed by the cloning technique and the accuracy of these constructs was confirmed by restriction enzyme digestion and DNA sequencing. Secondly, plasmids of pCI-S and pcDNA3.1-S were transferred into HepG2 and COS-7 cells, respectively by means of cationic liposome. HBsAg transient expression was assayed by ELISA in cell culture supernatants and cell lysates. Thirdly, plasmids were injected into quadriceps muscles of BALB/C mice and serum samples were obtained from individual immunized or control mice 4 weeks after injection and boost injection, respectively. Anti-HBs were assayed in mice sera by ELISA. HBsAg-specific CTL responses of spleen cells from immunized mice were tested by the LDH method.

RESULTSPlasmids of pCI-S and pcDNA3.1-S allowed HBsAg transient expression in cell culture supernatants and cell lysates of HepG2 or COS-7 cells. Intramuscular immunization of BALB/C mice with plasmids of pCI-S or pcDNA3.1-S elicited the antibody and cytotoxic T lymphocyte responses to HBsAg.

CONCLUSIONSThe vectors used in this study are effective to induce prime antibody and HBsAg-specific-cytotoxic T lymphocyte responses to HBsAg in mice after intramuscular immunization.

Animals ; COS Cells ; Cloning, Molecular ; DNA, Viral ; genetics ; Eukaryotic Cells ; metabolism ; Female ; Gene Expression ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis, Viral, Animal ; immunology ; prevention & control ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Cells, Cultured ; Vaccines, DNA ; genetics ; immunology ; Viral Vaccines ; genetics ; immunology

OBJECTIVE:To establish HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry for rapid determination of aconitine,mesaconitine,hypaconitine,benzoylaconitine,benzoylmesaconine and benzoylhypacoitine in rat plasma. METHODS:Internal standard lappaconitine was added into plasma sample,and methanol precipitated protein was used for pretreatment. HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry was adopted. HPLC condition was as follows as Sinochrom ODS-BP C18column,mobile phase consisted of acetonitrile-1% formic acid solution(50:50,V/V),the flow rate of 0.6 mL/min,sample size of 10 μL,column temperature of 25 ℃,automatic sampler temperature of 4 ℃. Mass spectrum scanning mode was full ion monitoring model,positive ion acquisition,mass charge ratios(m/z)of ion were 646.32(aconitine), 632.30(mesaconitine),616.31(hypaconitine),604.31(benzoylaconitine),590.29(benzoylmesaconine),574.30(benzoylhypacoitine), 585.31(internal standard). Six male Wistar rats were collected and given single dose of total alkaloid extract of Aconitum carmichaeli(4 mg/kg)intragastrically. Blood samples were collected before medication(0 h)and 0.5,0.75,1.25,1.5,2,4,6, 8,10,24 h after medication. Plasma concentration was determined and pharmacokinetic parameters were calculated by using PK-Solver V2.0 software. RESULTS:The linear range of 6 kinds of aconitum alkaloids in plasma were 0.1-10 μ g/L(r>0.992). The limit of quantitation was 0.1 μ g/L. Average recovery was higher than 75%,RSDs of intra-day and inter-day,matrix effects,stability test were all lower than 15%. The tmaxof 6 kinds of aconite alkaloids were about 1.2 h;t1/2were about 10 h;cmaxof monoestertype aconite alkaloids were higher than those of diester-type aconite alkaloids. CONCLUSIONS:Established HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry is accurate,sensitive,simple and rapid, and can be used for plasma concentration monitoring of 6 kinds of aconitum alkaloids.

Objective:To analyze the genotype of hereditary eye diseases with early-onset high myopia (eoHM) and its relationship with phenotype.Methods:The families with eoHM were collected in Ningxia Eye Hospital from January 2019 to June 2020.The medical records of the probands and their family members were inquired and recorded in detail, and the relevant ocular examinations were performed.Peripheral venous blood samples were collected from patients and their family members, and whole-genome DNA was extracted.Sequence capture sequencing technology was applied to screen for disease-causing gene mutations in probands.The detected suspected pathogenic variants were verified by Sanger sequencing and were analyzed by family cosegregation analysis.According to ACMG guidelines, the pathogenicity of novel variants was evaluated.The original literature about hereditary eye diseases with eoHM was searched to analyze the relationship between mutated genes and clinical phenotype.This study protocol adhered to the Declaration of Helsinki.All subjects or their guardians were informed of the purpose and procedure of the study and signed the informed consent form.The study protocol was approved by the Ethics Committee of the People's Hospital of Ningxia Hui Autonomous Region (No.2016018).Results:A total of 20 eoHM families were collected, among which pathogenic variants associated with inherited eye diseases were detected in 8 families.Of the 8 probands, two were diagnosed with familial exudative vitreoretinopathy, one with X-linked retinitis pigmentosa, one with congenital stationary nightblindness, one with Stickler syndrome, one with achromatopsia, one with Leber congenital amaurosis, and one with gyrate atrophy of the choroid and retina.The first diagnosis age of the 8 probands was 4-7 years old, and they were all diagnosed as high myopia, with a refractive status ≤-6.00 DS.Genetic tests showed that the 8 probands carried a heterozygous variant c. 313A>G (p.M105Val) in FZD4 gene, a heterozygous variant c. 14_15insAAGA (p.Asp5fs *) in TSPAN12 gene, a heterozygous frameshift variant c. 2234_2237del (p.Arg745fs) in RPGR gene, a compound heterozygous variant of c. 481C>T (p.Gln161Ter *) and c. 355>T (p.Arg119Cys *) in GPR179 gene, a frameshift variant c. 1659_1660insACGGTGACCCTGGCCGTCCTGG (p.Pro554fs *) in COL2A1 gene, a compound heterozygous variant of c. 1811C>T (p.Thr604Ile *) and c. 967G>A (p.Gly323Ser) in PDE6B gene, a compound heterozygous variant of c. 604_619delTCCACGGCACTCAGGG (p.Ser202fs *) and c. 995G>C (p.Arg332Pro) in GUCY2D gene, a homozygous variant c. 772C>T (p.Pro241Leu) in OAT gene.Seven of them were novel variants.Compared with the previous literature, the clinical and gene phenotypes of the 8 families were analyzed in detail in this study, which provided the basis for the diagnosis of hereditary eye diseases with eoHM. Conclusions:EoHM is closely related to some hereditary eye diseases, which may be the reason for the early diagnosis of children and an important clue for clinicians to detect potential hereditary eye diseases.Further clinical evaluations of ocular structure and function as well as genetic screening in children with eoHM are recommended.

Novel Oncogene with Kinase-domain (NOK) is a novel tumor-related gene, coding receptor like protein with a kinase domain. Overexpression of NOK leads to tumorigenesis and metastasis. To further study NOK function in physiological condition, it is necessary to prepare the anti-NOK antibody. In this report, GST fusion protein was adopted to prepare polyclonal antibodies against hNOK. The result showed that the antibodies we generated is with a very high titriation, and can be used for examination of NOK protein by Westernblot. Furthermore, the antibodies were used for immunohistochemistry in lung cancer tissues, and the results demonstrated high expression of hNOK in the tumor tissues. The antibody of hNOK we generated can serve as a diagnostic method for the lung cancer.
Animals ; Antibodies ; genetics ; metabolism ; Antibodies, Monoclonal ; biosynthesis ; genetics ; Biomarkers, Tumor ; genetics ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; genetics ; metabolism ; Mice ; Oncogene Proteins ; biosynthesis ; genetics ; immunology ; RNA, Messenger ; biosynthesis ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; immunology