Background and purpose:miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods:Quantitative real-time palymerase chain reaction (qRT-PCR) method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expres-sion of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays.Results:qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that theD values were signiifcantly lower than the control group, (P<0.05). After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was (105±7) and (107±13), respectively, which are signiifcantly different from the control group (213±11), indicating invasion of SKOV3 cells signiifcantly slowed down (P<0.05).Conclusion:miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.

Objective To discuss the diagnostic value of contrast-enhanced transvaginal sonography TVS in the diagnosis of myometrial invasion grade of endometrial cancer. Methods Total twenty-eight cases were studied by contrast-enhanced TVS,which were proved pathologically endometrial cancer in our hospital. The diagnosis of myometrial invasion grade of endometrial cancer by contrast-enhanced TVS were compared with pathologic results based on FIGO. Results The total coincidence of contrast-enhanced transvaginal sonography in detecting the depth of myometrial invasion was 78.6%. In the evaluation of no myometrial invasion,it showed the sensitivity of 71.4%,specificity of 85.7% and coincidence of 62.5%. In evaluation of superficial myometrial invasion,the sensitivity,specificity and coincidence was 73.7% ,77. 8% ,87. 5% ,respectively. The sensitivity in detecting deep invasion was 100. 0% ,the specificity was 96.0% ,while coincidence was 75.0%.There was no significant difference among the coincidence of various depth of myometrial invasion by contrast-enhanced TVS. Conclusion Contrast-enhanced TVS was valuable in the diagnosis of myometrial invasion grade of endometrial cancer.

Objectives This study was designed to observe the correlation of the levels of serum N-terminal pro-brain natriuretic peptide(NT-proBNP) and high sensitivity C-reactive protein(hs-CRP) between the 3rd day after acute myocardial infarction(AMI) and after 3 months of left ventricular remodeling(LV remodeling) and to establish the predictive levels of NT-proBNP,hs-CRP for LV remodeling after AMI.Methods The blood samples from 106 patients with the first AMI and echocardiography were examined on the 3rd day and after 3 month.LV remodeling was estimated by the changes of left ventricular end-diastolic volume(LVEDV) during the 3rd day and after 3 month.Based on the change of LV remodeling,the 106 patients were divided into two groups:LV remodeling group(defined as the change rate of LVEDV more than 20%) and non-LV remodeling group.Results The correlation coefficients of the change of LVEDV were 0.706 for serum NT-pro BNP and 0.596 for hs-CRP.With a cutoff value of 0.20,the area under the ROC curve(AUCs) was(0.892) for NT-proBNP and 0.825 for hs-CRP.There was no statistically difference between NT-proBNP and hs-CRP.The ROC plot indicated that NT-proBNP was superior to hs-CRP for predicting LV remodeling.Conclusion The level of serum NT-proBNP appropriately reflects LV remodeling after AMI and shows more efficacious than the level of serum hs-CRP.The level of serum NT-proBNP on the 3rd day after AMI and LV remodeling were positively correlated with the level after 3 months.

Objective To investigate the effect of the ubiquitin-proteasome pathway inhibitor MG132 on the natural resistance to cisplatin in the human cervical cancer line (HCE1) muhicellular spheroid (HCE1/MCS) model and to probe it if MG132 could reverse the HCE1/MCS resistance to cisplatin, as well as the possible mechanism of drug resistance.Methods (1) HCE1/MCS was obtained using liquid overlay and rotating technique.(2)Four groups were established (MG132 group, cisplatin group, MG132 + cisplatin group, the control group).Cell viability were measured by trypan blue exclusion assay.Cell cycle and apoptosis were detected by flow cytometry.(3) The expression of nuclear factor (NF) kB of both HCE1 monolayer cells (HCEI/MC) and HCE1/MCS was detected by western blot, and the expression of B cell lymphoma/leukemia-2 (bcl-2) was detected by immunohistochemistry.Results (1) HCE1/MCS was established successfully.(2) Cell inhibited rate of HCE1/MC and HCE1/MCS was: in MG132 group, (11.67 ± 2.34) % vs (10.78 ± 1.17) % (P > 0.05) ; in MG132 + cisplatin group, (92.67 ± 2.52)% vs (91.33 ±2.18)% (P>0.05); in cisplatin group, (45.01±7.44)% vs (9.45±5.98)% (P<0.05).(3)The rate of apoptosis of HCE1/MC and HCE1/MCS were: in MG132 group, 8.14% and 5.97% ; in MG132 + cisplatin group, 99.01% and 95.22% ; in cisplatin group, 33.61% and 0.88%.(4)The expression level of NF-kB and the high expression rate of bcl-2 were: in HCE1/MCS of control group, 0.67 and 60% ; in HCE1/MCS of cisplatin group, 0.85 and 83% ; in HCE1/MCS of MG132 group, 0.39 and 20% ; in HCE1/MCS of MG132 + cisplatin group, 0.47 and 33%.Conclusions (1) HCE1/ MCS present natural resistance to cisplatin and may become a good model for the study of cervical cancer drug resistance in vitro.(2) MG132 could induce the inhibition and apoptosis of HCE1/MCS cells and partially reverse the natural resistance of HCE1/MCS to cisplatin, of which partially reverse the natural resistance may be in relation to the down-regulation of NF-kB and bcl-2 expression.

BACKGROUND:Posterior lamina resection often causes loss of spinal stability, so screw rod internal fixation technology is needed to maintain the stability of lumbar spine. Finite element analysis can be used to simulate the stress distribution of the spine and internal fixation system after spinal surgery. OBJECTIVE: To build three-dimensional finite element model of spinal L1 to L3, analyze the spinal stability and stress distribution after the total laminectomy and insertion of bilateral pedicle screw using finite element method. METHODS: L1-L3 CT data could be colected from an adult healthy male volunteer. Mimics14.01, 3-matic(V6.0) and Ansys 15.0 could be used to set up the intact lumbar spine finite element model of L1-L3 (group A), the L1-L3 finite element model after L2 total laminectomy (group B), and the finite element model of L2 total laminectomy and insertion of bilateral pedicle screw (group C). We used software to simulate flexion, extension, lateral bending and axial rotation, and three kinds of models received finite element analysis. RESULTS AND CONCLUSION: (1) Based on the maximum of Von Mises under different motion states, the maximum stress was significantly lower in group A than in group B (P< 0.05). The maximum stress was significantly lower in group B than in group C (P < 0.05). (2) Based on the total deformation under different motion states, the total deformation was significantly lower in group A than in group B (P < 0.05). The total deformation was significantly lower in group C than in groups A and B (P < 0.05). (3) After the total laminectomy, vertebral body stress increased, especialy in the lamina, pedicle and joints. The range of motion of the vertebral body increased, which influenced the stability of the vertebral body. Internal fixation could decrease range of motion. Stress concentrated on the screw. Stress on the vertebral plate and pedicle decreased. The stability of vertebral body increased. Excessive stress concentrated on screw system wil increase the risk of screw breakage.

The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 10(4). Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70 degrees C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471lle. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Va1596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme.
Aflatoxin B1 ; antagonists & inhibitors ; chemistry ; Amino Acid Substitution ; Directed Molecular Evolution ; Enzyme Activation ; Enzyme Stability ; Multienzyme Complexes ; genetics ; metabolism ; Mutant Proteins ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; methods ; Protein Engineering

Objective To explore the relationship between serum retinol binding protein 4 and serum Lipasin levels and vascular complications in gestational diabetes mellitus(GDM).Methods From Jan.2016 to Jan.2018,80 pregnant women with gestational diabetes diagnosed as GDM in Wenzhou Central Hospital of Zhejiang Province were selected as the study group.They were divided into two groups according to whether they had vascular lesions.Group A included patients with gestational diabetes mellitus complicated with vascular complications and group B included patients without vascular complications.Forty healthy pregnant women were selected as the control group(group C).The levels of fasting plasma glucose (FPG),fasting serum insulin (FINS),homeostatic model assessment of insulin resistance (HOMA-IR),serum retinol binding protein 4 (RBP4) and serum Lipasin were compared among three groups of pregnant women.The vascular complications of GDM were analyzed.Results The levels of FPG,FINS and HOMA-IR in gestational diabetes mellitus pregnant women were higher than those in healthy pregnant women (P＜0.05),and those in group A were higher than those in group B (P＜0.05).The levels of RBP4 and Lipasin in serum of pregnant women with GDM were higher than those of healthy pregnant women (P＜0.05),and those of group A were higher than those of group B (P＜0.05).Spearman univariate correlation analysis showed that serum RBP4 levels were positively correlated with FPG and FINS(P＜0.05),and serum Lipasin levels were positively correlated with FPG and FINS (P＜0.05).Logistic regression analysis showed that the levels of FPG,FINS,RBP4 and Lipasin increased,which were independent risk factors for diabetic retinopathy (P＜0.05).Conclusion In GDM with vascular complications,the serum RBP4 and Lipasin levels are higher,which are independent risk factors for vascular complications in GDM,and may participate in the occurrence and development of vascular complications in gestational diabetes mellitus.

Injection fear is widespread in the population, which can cause patients to tolerate or avoid injection, reduce treatment compliance, and increase the burden of healthcare. Choosing appropriate injection fear assessment tools in clinical practice is helpful to understand the degree, psychological characteristics and influencing factors of individual injection fear. In this paper, the contents, characteristics and application methods of fear of injection assessment tools at home and abroad are reviewed, in order to provide reference for the application and development of fear of injection assessment tools for medical staff.

Objective:To investigate the effect of miR-503 targeting CBX2 on drug resistance of breast cancer MDA-MB-231 cells and its potential mechanism.Methods:miR-con group, miR-503 group, si-con group, two groups of si-chromosome homologues (CBX), anti-miR-con group, anti-miR-503 group, miR-503+pcDNA group, miR-503+pcDNA-CBx2 group were set up. Real-time quantitative fluorescence polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-503 and CBX2 mRNA. Western blot was used to detect protein expression. Cell activity was detected by MTT assay. The targeted regulatory relationship was detected by double luciferase assay.Results:Compared with normal breast cells HBL-100 (1.02±0.09), the expression level of miR-503 in breast cancer cells MCF-7 (0.41±0.05), MDA-MB-231 (0.25±0.03) and BT474 (0.35±0.04) was significantly decreased. The expression levels of CBX2 mRNA in MCF-7, MDA-MB-231 and BT474 cells were (4.02±0.35), (4.62±0.36) and (3.47±0.33), respectively. The expression levels of CBX2 protein in MCF-7, MDA-MB-231 and BT474 cells were (0.64±0.07), (0.74±0.05) and (0.68±0.06), respectively. The mRNA and protein contents of CBX2 in normal breast cells HPL-100 were (1.01±0.08) and (0.40±0.04), respectively, and the expression of CBX2 in breast cancer cells was significantly higher than that in normal breast cells ( P<0.05). Overexpression of miR-503 (3.64± 0.30) and silting of CBX2 inhibited proliferation, migration and invasion of MDA-MB-231 cells, and inhibited CBX2 (0.26±0.03), cyclin-dependent kinases, CDK) 4 (0.32± 0.03), Cyclin (CCN) D1 (0.58±0.03), matrix metalloproteinases (matrix metalloproteinases), MMP-2 (0.32±0.03) and MMP-9 (0.32±0.04) ( P<0.05). miR-503 targeted the expression of CBX2, and overexpression of CBX2 (0.75±0.03) could reverse the proliferation and drug resistance of miR-503 to MDA-MB-231 breast cancer cells. Conclusion:miR-503 may inhibit the proliferation, migration and invasion of breast cancer cells by down-regulating the expression of CBX2.

Porcine interferon-alpha (pIFN-alpha) fermentative production by recombinant Pichia pastoris was carried out in a 10-L bioreactor to study its metabolism changes and effects on fermentation under different inducing strategies, by analyzing the change patterns of the corresponding metabolism and energy regeneration. The results show that the specific activities of alcohol oxidase (AOX), formaldehyde dehydrogenase (FLD) and formate dehydrogenase (FDH) largely increased when reducing temperature from 30 degrees C to 20 degrees C under pure methanol induction, leading significant enhancements in methanol metabolism, formaldehyde dissimilatory energy metabolism and pIFN-alpha antiviral activity. The highest pIFN-alpha antiviral activity reached 1.4 x 10(6) IU/mL, which was about 10-folds of that obtained under 30 degrees C induction. Using methanol/sorbitol co-feeding strategy at 30 degrees C, the major energy metabolism energizing pIFN-alpha synthesis shifted from formaldehyde dissimilatory energy metabolism pathway to TCA cycle, formaldehyde dissimilatory pathway was weakened and accumulation of toxic intermediate metabolite-formaldehyde was relieved, and methanol flux distribution towards to pIFN-alpha synthesis was enhanced. Under this condition, the highest pIFN-alpha antiviral activity reached 1.8 x 10(7) IU/mL which was about 100-folds of that obtained under pure methanol induction at 30 degrees C. More important, enhanced pIFN-alpha production with methanol/sorbitol co-feeding strategy could be implemented under mild conditions, which greatly reduced the fermentation costs and improved the entire fermentation performance.
Animals ; Energy Metabolism ; Fermentation ; Interferon-alpha ; biosynthesis ; genetics ; Methanol ; pharmacology ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Sorbitol ; pharmacology ; Swine