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Objective To explore the mechanism of human umbilical cord mesenchymal stem cell (HUC-MSC) alleviating ischemia-reperfusion injury (IRI) of liver cells through mitochondrial transfer. Methods Normal human liver cell line L02 was divided into the blank control group, oxygen-glucose deprivation (OGD) group, experimental control group, and L02 and HUC-MSC co-culture group (L02+HUC-MSC group). L02+HUC-MSC group was further divided into 10:1 co-culture subgroup (group A), 4:1 co-culture subgroup (group B), 2:1 co-culture subgroup (group C), 1:1co-culture subgroup (group D) and 1:2 co-culture subgroup (group E) according to different co-culture ratio of L02 and HUC-MSC. The apoptosis rate and relative reactive oxygen species (ROS) level of L02 cells were detected by flow cytometry. The MitoTracker positive rate of L02 cells was detected by flow cytometry. The mitochondrial transfer from HUC-MSC to L02 cells was observed by laser confocal microscope. Results The apoptosis rate and relative ROS level of L02 cells in the OGD group were significantly higher than those in the blank control group (both P < 0.05). Compared with the OGD group, the apoptosis rates of L02 cells in group B, C, D and E were significantly decreased (all P < 0.05), and the relative ROS level of L02 cells in group E was significantly declined (P < 0.05). The MitoTracker positive rate of L02 cells did not significantly differ between group A and experimental control group (P>0.05), whereas the MitoTracker positive rates of L02 cells in group B, C, D and E were significantly higher than that in the experimental control group in a concentration-dependent manner (all P < 0.05). Under the laser confocal microscope, mitochondrial transfer fromHUC-MSC to L02 cells could be observed through tunneling nanotube (TNT). Conclusions HUC-MSC may alleviate cell apoptosis and reduce ROS level of liver cells after IRI via direct mitochondrial transfer between cells.
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Dravet syndrome ( DS) , known as severe myoclonic epilepsy of infancy, is a devastating disorder characterized by intractable epilepsy and poor neurodevelopmental outcome. Seizures are refractory to conventional antiepileptic therapy, therefore resulting in heavy psychological pressure and burden. Stiripentol ( STP) is a novel antiepileptic drug, which has been proposed to achieve better seizure control in DS. It acts as a direct GABA receptor agonist by increasing the GABAergic transmission and by prolongating the opening peri-od of the receptor dependent chloride channels. In addition, STP inhibits several isoenzymes of the cytochrome P450 system in the liver involved in the metabolism of other antiepileptics, thus potentiating their effects. By summarizing the relevant researches at home and abroad, we describe an on-going work in the anticonvulsant mechanisms and efficacy of STP, to provide an alternative treatment of DS.
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Objective To evaluate the effect of different doses of D-galactosamine (D-Gal) on non-human primate cynomolgus monkey and to establish a monkey model with different degree of acute liver failure (ALF). Methods Twelve cynomolgus monkeys were evenly divided into the low-, medium- and high-dose groups (n=4) with a dosage of 0.23, 0.25 and 0.27 g/kg, respectively. In each group, the corresponding dose of D-Gal solution was injected into the monkeys through the forearm vein at one time in a sober state (without anesthesia). The survival time of the cynomolgus monkeys was recorded. Digestive tract and hepatic encephalopathy symptoms were observed. Vital signs were measured at 0 h before and 12, 24, 36, 48, 60, 72, 96, 120 and 144 h after D-Gal administration. Alanine transaminase (ALT), total bilirubin (TB), prothrombin time (PT), blood ammonia and other parameters were detected from the blood samples. The liver tissues were prepared for hematoxylin-eosin (HE) staining to observe the pathological changes. Results All cynomolgus monkeys in the low-dose group survived and transient liver injury was noted without the hepatic encephalopathy symptoms. At 60 h after D-Gal administration, the liver function and coagulation indexes reached the peak, gradually recovered and then basically returned to the normal range at 120 h. In the medium-dose group, the course of disease was relatively slow and gradually recovered after the appearance of severe liver damage and hepatic encephalopathy symptoms and only one animal died. All cynomolgus monkeys in the high-dose group died after developing hepatic encephalopathy symptoms and severe liver damage with a mean survival time of (72±13) h. Pathological examination of liver tissue demonstrated that scattered liver cell necrosis and inflammatory cell infiltration were observed in the liver tissues of the low-dose group. In the medium- and high-dose groups, the hepatic lobule structure was not clear, and the liver cell necrosis in flakes accompanied by evident hemorrhage were documented. Conclusions The D-Gal dosage in the medium- and high-dose groups meet the standards of the ALF model. The degree of ALF in the medium-dose group is relatively slight, which is beneficial to the implementation of liver transplantation. ALF in the high-dose group is relatively severe, which is suitable for the evaluation of the clinical efficacy of therapeutic options.
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OBJECTIVE@#To report on a case of 10p15.3 microdeletion syndrome and to explore its clinical and molecular characteristics.@*METHODS@#The patient was subjected to whole exome sequencing (WES), with his clinical features discussed in the light of literature review.@*RESULTS@#The patient presented with global developmental delay, hypotonia, autistic-like traits, mild facial dysmorphism and other features including short stature, small hands and feet, congenital heart disease and feeding difficulty. WES has detected deletions of ZMYND11, DIP2C, LARP4B, TUBB8, GTPBP4, IDI2, IDI1, WOR37 and ADARB2 genes on the short arm of chromosome 10. Among these, ZMYND11 gene been previously associated with intellectual disability.@*CONCLUSION@#The patient's phenotype was closely correlated with that of 10p15.3 microdeletion syndrome. Haploinsufficiency of the ZMYND11 gene may underlie the manifestations of 10p15.3 microdeletion syndrome.
Subject(s)
Humans , Carrier Proteins , Chromosome Deletion , Chromosomes, Human, Pair 10 , Exome , GTP-Binding Proteins , Intellectual Disability , Nuclear Proteins , Phenotype , Tubulin , Exome SequencingABSTRACT
Objective The objective of this study was to investigate the effect of Wnt pathway inhibitor-ETC-159 on pro-liferation and migration of oral squamous cell carcinoma(SCC-15)cells and to explore its mechanism.Methods SCC-15 cells were treated with DMSO and ETC-159 for 12 h or 24 h.Cell proliferation was detected by CCK8 kit.Transwell assay was used to de-tect the ability of cell migration.Western blotting was used to detect cell migration related to proteins i.e.Wnt3a and β-catenin,pro-liferation and migration related proteins i.e.c-Myc,cyclin D1,CD146.Results After treated with ETC-159 for 12 or 24 h,the proliferation,migration and expression of Wnt3a,β-catenin,c-Myc,cyclin D1 and CD146 in SCC-15 cells were significantly de-creased when compared to the DMSO group(P<0.05).Conclusion Wnt signaling pathway inhibitor-ETC-159 can inhibit the proliferation and migration of SCC-15 cells by decreased levels of c-Myc,cyclin D1 and CD146.
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Ketogenic diet(KD)has an auxiliary treatment for refractory epilepsy. It is recently demonstrated by clinical practice and researches that KD is effective in some children with refractory epilepsy. Despite much progress has been made on the anticonvulsant mechanisms of KD,no biological target related to clinical efficacy has been found so far. The biochemical characteristics of KD and its metabolic changes are discussed(including the roles of neuronal γ -aminobutyric acid,glutamate,ATP sensitive potassium channels and purine metabolism,and so on). The mechanism and recent progress of KD in the treatment of refractory childhood epilepsy are reviewed.
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Objective To investigate the risk factors of prehypertension among postmenopausal women.Methods2 592[(43±12)years old] health women were collected as the research object in Saihan District of Hohhot in Inner Mongolia Autonomous Region in April 2015,including 697[(58±6)years old] postmenopausal subjects and 1 895[(37±8)years old] premenopausal subjects.T test was used to compare means of blood pressure, fasting blood glucose (FBG), blood lipid, and body massive index (BMI) between postmenopausal group and premenopausal group and to compare prevalence of prehypertension, hyperglycemias, dyslipidemias, overweight, and obesity between two groups.Logistic Regression was implemented to analyze the relationship between different risk factors and prehypertension among postmenopausal women.Results Compared with premenopausal women, the systolic pressure, diastolicpressure, BMI, FBG, triacylglycerol (TG), low density lipoprotein (LDL) in postmenopausal women were significantly higher(P<0.05).Prevalence of prehypertension, impaired fasting glucose (IFG), diabetes, TG abnormalities, LDL abnormalities, overweight, and obesity in postmenopausal women were significantly higher than in premenopausal women(P<0.05).Age 55 to 59, 60 to 64 and above 65 years overweight, obesity, IFG and diabetes were independent risk factors of prehypertension among postmenopausalwomen.Conclusions Age 55 yearsand above, overweight, obesity, IFG and diabetes are the independent risk factors of prehypertension among postmenopausal women.
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Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.
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Background Lattice corneal dystrophy (LCD) is a progressive disease,whose clinical features are varied in different stages.It is rarely be reported that clinical findings of different stages and factors of promoting the occurrence and development on LCD in a family.Objective The aim of this study was to identify the characteristics of the pedigree and clinical features of different stages in a LCD family,and further to discuss its influence factors.Methods A cross-sectional study was performed in this study.A Chinese family with LCD was enrolled in Shenzhen Eye Hospital from 2015 to 2016.Questionnaires for disease-related history,visual acuity measurement,ocular anterior segment examination and color photography were carried out for all the members of the family.In addition,anterior segment OCT (AS-OCT),laser scanning confocal microscope and corneal endothelium microscope were used to observe the morphology of corneal stroma and changes of corneal endothelial cells.The pedigree chart was drawn by Cyrillic2.1 software and analyzed based on Mendel law.Results This family included 5 generations of 73 members.Patients with LCD were found in each generation with similar morbidity in different gender,which followed the law of autosomal dominant inheritance.Eleven patients were found in 49 members related with Ⅲ1 of this family with the prevalence rate of 22.45％ and onset age at 21-50 years old,and the course of disease was 3-34 years.All of the members had no systemic disease except for two patients (Ⅲ 1 and Ⅲ 5) with hypertension.In the early stage of LCD,some bifurcate striolae appeared in the patients' corneal stroma without symptoms for many years.In the progressive stage,there was corneal irritation symptom accompanying with vision's decrease in the eyes with LCD.The bifurcate striolae were increased,widened and interwoven into lattice lines that the boundaries gradually became fuzzy,then corneal macula was formed because of recurrent corneal infiltration,and eventually resulted in corneal leucoma.High reflection corresponding to the pathologic region was showed by laser scanning confocal microscope and AS-OCT.No significant differences were found in corneal endothelial cell density and the percentage of hexagonal cells between LCD patients and normal phenotype families (t =1.887,P=0.075;t=-0.719,P =0.481).Penetrating keratoplasty was performed in a patient with corneal opacity and serious corneal opacity occurred near the surgical incision one year after the surgery.One patient was diagnosed as LCD 2 years after laser assisted in-situ keratomileusis.One patient was a welder.Conclusions LCD is autosomal dominant inheritance in the family.The clinical manifestations of LCD in the early,progressive and late stage can be seen in the pedigree,which offers a reference for ophthahnologists.Corneal surgery and lesion may induce the onset or aggravation of LCD.
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Objective To optimize the extraction methods of mitochondrial genome DNA(mtDNA)of Oncomelania hupen-sis. Methods The pyrolysis,protein K variable-temperature digestion and high-concentration potassium acetate purification were applied to optimize the high-concentration-salt precipitation method,and then the optimized method was compared with two common extraction methods,the sucrose density gradient centrifugation method and traditional high-concentration-salt pre-cipitation method. The mtDNA samples were identified by using spectrophotometry,agarose gel electrophoresis and the amplifi-cation products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. Results The concen-tration and yield of the improved method was significantly higher than those of the traditional method(F=3032.65,10185.00, both P<0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein,meeting PCR,sequence analy-sis and other molecular biology research requirements. Conclusions The improved high-concentration-salt precipitation meth-od for isolating mtDNA is simple,and it has high yield and low cost. The extracted mtDNA can meet relevant analysis require-ments.
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Objective To detect the expression levels of adipose-specific phospholipase A2 (AdPLA) mRNA in orbital adipose tissue of thepatients with thyroid associated ophthalmopathy and the normals.Methods Sixteen patients with TAO of m level and stationary phase underwent orbital decompression,and 29 normals underwent ocular plastic surgery in Shenzhen Eye Hospital between August,2015 and October,2016.Orbital fat samples were collected from one eyes of these patients during surgery.The age,gender,height,weight,body mass index (BMI),exophthalmos degree,orbital fat of the patients with TAO and the normals were recorded and calculated.Using real time PCR,the AdPLA mRNA were detected from these orbital fat samples.Results There was no significant difference between the patients with TAO and the normals in age,gender,and BMI (all P ＞ 0.05).TAO group had more exophthalmos degree (20.406 ± 1.369)mm than the normals (14.207 ± 1.146) mm.TAO group had more orbital fat (32.162 ± 1.923) mL than the normals (24.279 ± 1.070) mL.The average expression level of AdPLA in patients with TAO was 0.039 42 ± 0.009 85,and 0.004 42 ± 0.001 36 in the normal.There was significant difference between two groups (P ＜ 0.05).Conclusion The patients with TAO of Ⅲ level and stationary phase have more exophthalmos degree and orbital fat than the normals.AdPLA mRNA is higher expressed in orbital adipose tissue of the patients with TAO of Ⅲ level and stationary phase than the normals.The high expression of AdPLA may reduce lipolysis in the orbital adipose tissue,lead to fat accumulation in orbits,and aggravate exophthalmos of patients with TAO.
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Objective To investigate the effect and mechanism of high-dose sirolimus (rapamycin) upon protecting the hepatic ischemia-reperfusion injury (HIRI) in aged mice. Methods Twenty C57BL/6 aged mice were randomly and evenly divided into the ischemia-reperfusion injury group (IRI group), low-dose rapamycin pretreatment group (rpm group), high-dose rapamycin pretreatment group (RPM group) and control group (Sham group) using the random number table method (5 mice in each group). In the Sham group, abdominal cavity was incised and sutured alone. In the other three groups, aged mouse 70% HIRI models were established. The ischemia time was 60 min. At preoperative 1 h, rapamycin at a dose of 1 mg/kg and 5 mg/kg was administered via intraperitoneal injection in the rpm and RPM groups. At 12 h post-reperfusion, hematoxylin-eosin (HE) staining was performed to observe the histological changes in the mouse liver. Suzuki grading method was adopted to evaluate the pathological score. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), tumor necrosis factor (TNF)-α and interleukin (IL)-10 and the LC3B-Ⅱ protein level in the liver tissues were quantitatively measured and statistically compared among different groups. Results HE staining of the liver tissues revealed normal liver tissues in the Sham group, severe liver cellular injury accompanied with a large quantity of inflammatory cellular infiltration in the IRI and rpm groups. Mild sinusoidal congestion and slight inflammatory cellular infiltration were observed in the RPM group. The pathological score was 5 (4-6) in the RPM group, significantly lower than 7 (5-8) and 8 (7-10) in the rpm and IRI groups (Z=-2.554 and -2.731, both P<0.05). In terms of postoperative liver function parameters, the AST level was (691±207) U/L in the RPM group, significantly lower compared with (2032±575) U/L and (1817±777) U/L in the IRI and rpm groups (t=4.90 and 3.13, both P<0.05). In the RPM group, the ALT level was measured as (996±584) U/L, considerably lower than (2992±992) U/L and (2373±687) U/L in the IRI and rpm groups (t=3.86 and 3.41, both P<0.05). The AST and ALT levels did not significantly differ between the IRI and rpm groups (both P>0.05). No statistical significance was identified in the TNF-α and IL-10 levels among different groups (all P>0.05). Western blot analysis revealed that the relative expression level of LC3B-Ⅱ protein in the liver tissue of the RPM group was significantly higher than those in the Sham, IRI and rpm groups (all P<0.05). Conclusions Administration of high-dose rapamycin exerts a protective effect upon HIRI probably through promoting cellular autophagy in aged mice.
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Objective To study the preventive effect ofω-6 soybean oil fatty emulsion on gastric ulcer caused by acetic acid in rat model, and investigate its mechanisms. Methods Thirty healthy rats were randomly and equally assigned to the following 3 groups:sham operation,gastric ulcer,andω-6 Soybean oil fatty emulsion group.The model was induced by acetic acid. Five days after the model was established successfully,rats in ω-6 soybean oil group received the treatment by tail intravenous injection with the dose of 10 mL.kg-1 .d-1 ,the sham operation group and gastric ulcer group were given the same dose of 0.9%sodium chloride solution.The rats were sacrificed at 10th day after the treatment.The pathological changes of rat gastric ulcer tissue were observed by HE staining, and the concentration of gastric acid was detected by acid-base neutralization method,as well as the activity of pepsin was detected by colorimetry.Serum NO concentration was detected with nitrate reductive enzymatic method, and the expression of EGFR in gastric mucosal was detected with immunohistochemical method. Results Gastric ulcer area inω-6 soybean oil fatty emulsion group (5.67±2.32 mm2) was significantly lower than that in gastric ulcer group(8.68±1.98 mm2). The concentration of gastric acid (1.70±0.53 mmol.L-1), activity of pepsin(23.12±6.97 U) and NO level (64.62±13.86μmol.L-1 ) inω-6 soybean oil fatty emulsion group were much lower than those in the model control group.While the expression of EGFR in gastric ulcer tissue was increased after treatment withω-6 soybean oil fatty emulsion. Conclusion ω-6 soybean oil fatty emulsion exerts significant promotion effect on the healing of gastric ulcer,and its mechanism might be related to inhibiting the level of gastric acid, pepsin and NO, while improving the protective effect of EGFR on gastric mucosa.
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Bring your own device (BYOD) has become popular as it empowers medical practitioners to use their own devices for communication,mobile rounds,real-time EMR query,selflearning and auxiliary diagnosis.Based on implementation experiences of BYOD at home and overseas healthcare institutions,this paper recommended domestic institutions on BYOD planning and deployment as follows:restructuring of their wireless networks and access control;better device supervision;open BYOD portal;and protection of both staff privacy and data security.This effort provides insights for BYOD development in China's healthcare institutions.
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Objective to investigate the protective effect of omega-3 fish oil fat emulsion on cyclophosphamide-induced gastric mucosal injury in mice. Methods Forty-five kunming mice were randomly divided into three groups as control,model,and omega-3 fish oil fat emulsion group(with 15 mice in each group). Mice of the two experiment groups were administrated with cyclophosphamide i.p. for 2 days to establish the damage model. then mice in omega-3 fish oil fat emulsion group received omega-3 fish oil fat emulsion at a dose of 15 mL/kg daily for 14 days. Meanwhile,the ani-mals in control group and model group were intravenously administered with the same volume of saline. the weight and food intake of the mice in each group were assessed daily. Five mice in each group were respectively sacrificed at day 1,day 7,day 14 after intravenous injection. Morphology of gastric mucosa was observed by HE staining and the activities of SOD and MAO in gastric mucosa were measured respectively by xanthine oxida-tion and ultraviolet spectrophotometry methods. Results Compared with the model group,the general status,nutritional status and the injury in stomach mucosa in omega-3 fish oil fat emulsion group were significantly improved. After 14 day′s treatment,the activities of SOD and MAO in gas-tric mucosa of mice in omega-3 fish oil fat emulsion group were significantly increased(P < 0.05)compared with model group. Conclusion omega-3 fish oil fat emulsion has a significant protective effect on the cyclophosphamide induced injury in gastric mucosa of mice,which may be related to the upregulation of MAO and SOD.
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Objective To determine anatomic parameters related to the surgery of lumbar cortical bone trajectory of normal adults and the feasibility of screw application in cortical bone and its specification and dimension.Methods Lumbar 3D image data of 80 adults between 18 and 40 years old (40 for each gender) were randomly selected as subjects of our research.With the help of the imaging software for 3D interactive viewing,the ideal starting point was supposed to be the junction of the center of the superior articular process and 1 mm below the inferior border of the transverse process.The diameter,length,lateral angle to the vertebral horizontal plane,cephalad angle to the vertebral sagittal plane of the trajectory and the horizontal distance from the insertion point to the lateral edge of the vertebral plate were measured.Differences of anatomic parameters for each gender,side and segment were analyzed.Results Differences of anatomic parameters on both sides of each segment had no statistical significance.At the ideal trajectory,the mean screw length was 37.56±2.41 mm at L1,38.72±2.36 mm at L2,39.51 ±2.51 mm at L3,39.78± 2.87 mm at L4 and 38.83±2.74 mm at L5.The mean screw diameters from L1 to L5 were 6.04±1.23 mm,6.17±1.24 mm,7.15±1.22 mm,8.02± 1.41 mm and 8.68± 1.42 mm respectively.However,differences of ideal entry angle of L1 to L5 had no statistical significance.The mean lateral angle from L1 to L5 were 8.46°±2.11°,9.37°±2.84°,9.62°±2.16°,9.53°± 1.98°,9.04°± 1.97°,while the mean cephalad angle to the vertebral sagittal plane from L1 to L5 were 26.49°±4.97°,25.94°±4.56°,26.42°±4.42°,26.29°±3.48°,26.89°±3.69°.The mean distance from the insertion point to the lateral edge of the vertebral plate gradually increased from L1 to L5,which were 1.19±0.75 mm,1.54±1.08 mm,2.01±1.45 mm,3.49±1.52 mm,4.47±1.32 mm respectively.The screw diameters of each segment for men were greater than those for women.Conclusion The common length of screw for lumbar cortical bone trajectory of normal adults might be from 35 mm to 40 mm.The safe upper limits of the screw diameter were 5.5 mm at L1,5.5-6.0 mm at L2,6.5-7.0 mm at L3,7.5 mm at L4 and 8 mm at L5.The average lateral angle of all 5 lumbar segments was 9.20°± 2.11° and the average cephalad angle was about 26.41°±4.22°.
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Objective To identify and analyze the DNA methylation induced pathogenic genes in breast cancer.Methods Using genomic data provided by the international cancer genome,we carried out systematic a-nalysis of the abnormal gene expression in breast cancer and epigenetic regulation mechanism by bioinformatics methods.Results We identified 428 genes with abnormal expressions in breast cancer by t test.Functional enrich-ment analysis revealed genes up-regulation in breast cancer were closely related to the cell cycle,and those down-regulated were significantly enriched in the hormone response function.Study on DNA methylation revealed that breast cancer showed an unique DNA methylation pattern.Further analysis reveals 23 breast cancer genes induced by abnormal DNA methylation.Conclusion DNA methylation can mediate the abnormal expression of breast canc-er genes,and is an important biological marker for early diagnosis and prognosis of breast cancer.
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Objective To investigate the curative effect and safety of haploidentical allogeneic cytokine-induced killer (CIK)in treatment of advanced hepatocellular carcinoma.Methods The peripheral blood mononuclear cell (PBMC) of the healthy immediate family members of 21 patients with advanced hepatocellular carcinoma (HCC) were collected,induced into haploidentical allogeneic CIK in vitro and transfused to the patients for 4 cycles.The curative effect and safety were assessed.Results The 21 patients were followed up for half a year.The survival rate was 81 % (1 7 /21 ).Among the 21 patients,1 1 cases were with stable disease and 1 0 cases were with progressive disease (including 4 dead cases).Six patients developed fever of different degrees during the treatment and one patient developed rash.The platelet counts of the patients at the fourth cycle after the treatment decreased compared with that before the treatment ,with significance difference (P <0.05).The difference in leukocytes,neutrophils,lymphocytes,hemoglobin,liver function and renal function at the first and fourth cycle after the treatment had no statistical significance (all in P >0.05 ).Conclusions Haploidentical allogeneic CIK in treatment of advanced HCC may effectively improve the quality of life and the adverse reactions are tolerable,which is a relatively safe therapy.
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Objective To investigate the correlationship between DNMT3a, DNMT3b protein expressions and the state of promoter methylation of ERα gene and ERα protein expression in the development of sporadic breast cancer. Methods A total of 180 patients with sporadic breast cancer and 30 patients with breast fibroadenoma were included in this study. The expressions of DNMT3a and DNMT3b protein were detected by immunohistochemical method. The state of promoter methylation of ERα gene was detected by methylation specific PCR in 97 patients with sporadic breast cancer. Results There were no significant differences in positive expression rates of DNMT3a and DNMT3b protein between breast fibroadenoma and breast cancer. There were higher expression levels of DNMT3a and DNMT3b in breast cancer patient of Ⅲ～Ⅳstages than those of Ⅰ～Ⅱstages. The expression of DNMT3a was significantly higher in patients with lymph node metastasis than that of patients without lymph node metastasis (P＜0.05). Of 97 cases of breast cancer patients, ERα gene promoter methylation occurred in 39 cases (40.2%). The positive expression of DNMT3a protein was positively correlated with the ERα gene methylation (rS=0.250). The DNMT3a protein expression showed a significant influence to the overall survival (OS) in patients of breast cancer (P=0.035), no significant influence to the disease-free survival (DFS) (P=0.064). DNMT3b protein expression showed no significant influence to OS and DFS of patients with breast cancer (P=0.914 and 0.961). Conclusion The positive expressions of DNMT3a and DNMT3b are correlated with the invasion, metastasis and poor prognosis of sporadic breast cancer. DNMT3a was positively correlated with the state of ERα gene promoter methylation. The inhibition of DNMT3a and DNMT3b may have advantages in the prevention and treatment of sporadic breast cancer.
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Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.