In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.

Human parvovirus B19, generally referred to as B19 virus, has been closely associated with a variety of different autoimmune diseases due to the features of its capsid protein. B19 virus VP1 unique region protein that has sites of phospholipase A2 and several antigenic determinants is exposed on the outside of the viral capsid protein. As a result of that, B19 virus VP1 unique region protein can stimulate the host to produce autoantibodies, which induces and/or aggravates the autoimmune diseases. The biological characteristics of B19 virus VP1 unique region protein and its relationships with autoimmune diseases are de-scribed in this review based upon the published literatures and the work achieved by our research team. This review will be helpful to the prevention and treatment of B19 virus infection.

Objective To optimize the formula of konjac glucomannan-paeonol matrix tablets. Methods The formula of paeonol matrix tablets was optimized by the orthogonal design with the accumulative release rate in vitro as index, with the viscosity of konjac glucomannan ( KGM) , the amount of KGM and lactose as influence factors. Results The optimized formula was as follows:the viscosity of konjac glucomannan was 20 000 mPa·s, KGM 30%, lactose 20% and the release in vitro fit into the Higuchi equation. Conclusion The formula of the paeonol matrix tablets is reasonable and the tablets have well release effect in vitro.

Objective To study the feasibility of testing three disease-causing genes for Alport syndrome,COL4A3,COL4A4 and COL4AS,in diagnosing patients with sporadic Alport syndrome by using targeted capture and next generation sequencing.Methods The clinical data of a 9-year-old boy suspected with Alport syndrome were collected.Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes of the patient and his parents,respectively.Targeted capture and next generation sequencing and Sanger sequencing were applied to analyze the mutations in the 3 disease-causing genes.Clinical data of cases reported already with autosomal recessive Alport syndrome caused by the mutations in the COL4A4 gene were summarized.Results The patient was presented with neither family history of hematuria nor chronic renal failure.Haematuria and proteinuria were found at the age of 1 year.The patient presented with episodes of macrohaematuria and gradually developed nephrotic-level proteinuria.At the age of 8 years 7 months,bilateral sensorineural hearing loss was diagnosed.So a probable diagnosis of Alport syndrome was postulated.A compound heterozygous pathogenic mutations of 3578-1G ＞ A and 3967 C ＞ T(Q1323X) were identified in the COL4A4 gene in the patient.The mutation of 3578-1G ＞ A was inherited from his father,and the mutation of Q1323X from his mother.The patient was decisively diagnosed with autosomal recessive Alport syndrome.Conclusions The test of COL4A3,COL4A4 and COL4A5 genes can help diagnose patients with sporadic Alport syndrome by using targeted capture and next generation sequencing.

Interleukin-8 is CXC chemokine that is initially discovered using chemotaxis and the activation of neutrophils and induces the migration and proliferation of smooth muscle cells. Interleukin-8 is a potent angiogenic factor that may play a role in atherosclerosis. To establish the temporal correlation between IL-8 expression and plaque development, we examined the expression during atherosclerosis of hyperlipemia rabbits using immunohistochemistry, ELISA, in situ hybridization. By location of immunohistochemistry, the expression of IL-8 protein increased obviously in intima of hyperlipemia rabbits at 8 and 12 week. Quantitative analysis of the expression of IL-8 Immunohistochemistry indicated that positive area of AS model was 4.48 times and 8.76 times that of control group at 8 and 12 week. The valuation of IOD of AS model was 4.16 times and 4.36 times that of control group at 8 and 12 week. By specific ELISA, the ratio of the IL-8 protein to total protein of AS model was 1.84 times and 2.06 times that of control group at 8 and 12 week. By location of in situ hybridization, positive location was strong in intima of hyperlipemia rabbits at 8 week. We observed the dynamic alteration of interleukin-8 protein and gene expression in atherosclerotic lesions of hyperlipemia rabbits with establishing model. Interleukin-8 protein and gene expression was up-regulation in the development of fatty streaks in hyperlipemia rabbit.
Animals ; Aorta ; metabolism ; Atherosclerosis ; etiology ; metabolism ; Hyperlipidemias ; complications ; Interleukin-8 ; biosynthesis ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits

To establish and observe rabbit hyperlipemia and atherosclerosis model, we combined the method of high-lipid diet and immuoreactive injure. We divided 45 New Zealand rabbit into control group and high-lipid group. According to the time (4, 8, 12 weeks) of established model, the control group and high-lipid group were divided into 3 groups respectively. The blood-lipid, the hemodynamics parameter and vascular intima were determined and observed. The results showed: (1) After feeding 4, 8, 12 weeks, TC and LDL-C of the high-lipid group in the serum increased obviously. After feeding 8 week, TG of the high-lipid group began to increase obviously. HDL-C of the high-lipid group was higher than control group, but with a descendent trend. (2) Blood viscosity of the high-lipid group increased obviously under the 0.512 S(-1) and 5.96 S(-1) at 12 week. Blood flow increased obviously at 8 and 12 week. SBP increased evidently at 8 and 12 week. Alteration of the plasma viscosity and vascular diameter were not obvious. (3) By observing with optical microscope the intima of the control group is smooth. The intima of the high-lipid group has a light incrassation at 4 week. The intima of the high-lipid group has a obvious incrassation at 8 and 12 week. By observing with through transmission electron microscopy (TEM), the lipid vacuole is under the endothelium cell. (4) By adopting the immunohistochemistry, the foam cell that derived from smooth muscle cell were determined. We concluded that the blood lipid can have a prefigurative role in atherosclerosis; blood flow and blood pressure and blood viscosity increase at low shear rate in the course of the atherosclerosis; vascular intima is incrassate and the composition of the AS plaque is variational continually.
Animals ; Atherosclerosis ; blood ; pathology ; physiopathology ; Endothelium, Vascular ; pathology ; Hemodynamics ; Hyperlipidemias ; blood ; pathology ; physiopathology ; Lipids ; blood ; Male ; Rabbits

Fecal microbiota transplantation (FMT) has been reported to exert potential therapeutic value in gut microbiota dysbiosis.During FMT, the fecal material is transferred from a healthy donor into the gastrointestinal tract of a recipient via naso-gastric tube, enema, colonoscopy or oral capsules, aiming to reconstruct intestinal flora, ameliorate the dysbiotic state, control inflammation and modulate the immunity.FMT is recognized as a high-efficient and safe treatment option for recurrent clostridium difficile infection.It has been approved by the United States Food and Drug Administration for clinical use in the USA.FMT is a safe and effective treatment for patients with infla-mmatory bowel disease or autism spectrum disorder.

Objective: To investigate the prevalence and characteristics of pathogenic variants in complement genes in Han Chinese children with atypical hemolytic uremic syndrome (aHUS). Method: Eleven Han Chinese children with aHUS, including 9 boys and 2 girls aged between 1 year and 4 months and 13 years, were investigated in Department of Pediatrics, Fuzhou General Hospital, from November 1998 to February 2014. Analysis of variants of all the exons of 10 complement genes (CFH, MCP, CFI, C3, CFB, CFHR1, CFHR2, CFHR3, CFHR4 and CFHR5), including 25 bases from 3′ end and 25 bases from 5' end, was performed in the 11 cases by targeted sequence capture and next generation sequencing. Significant variants detected by next generation sequencing were confirmed by Sanger sequencing. To understand pathogenicity of variants found in the captured genes, we investigated genetic conservation by multiple protein sequence alignment among different species, and analyzed whether the variants were located in protein domains or not, and investigated functional significance by functional computational prediction methods. Result: Twenty-seven percent of Han Chinese children with aHUS carried pathogenic variants in the 10 complement genes. Pathogenic variant CFB 221G>A (R74H) was detected in Patient 3 and Patient 9, which was not found in parents of Patient 3′ , and was found in healthy father of patient 9. Pathogenic variant CFHR5 242C>T (P81L) was found in Patient 2, and was found in healthy father of patient 2. However, no pathogenic variants in genes CFH, MCP, CFI, C3, CFHR1, CFHR2, CFHR3 and CFHR4 were identified. Conclusion Pathogenic variants in the 10 complement genes were identified in 3/11 of Han Chinese children with aHUS in our study and CFB was the most frequently mutated gene.

Dravet syndrome is a rare and severe developmental epileptic encephalopathy with variable clinical phenotypes.Dravet syndrome is difficult to diagnose and treat, and related comorbidities have a profound impact on the long-term quality of life of patients and their parents.SCN1A is the main pathogenic gene of Dravet syndrome, and SCN1A mutations are found in more than 85% of the patients.In recent years, with the development of genetic testing technology and the accumulation of cases, the understanding of the characteristics of epileptic seizures, comorbidities and SCN1A gene mutation characteristics in Dravet syndrome has gradually deepened.In addition to conventional antiepileptic drugs, new antiepileptic drugs(cannabidiol, fenfluramine)have also shown good antiepileptic effects and are expected to become second-line drugs for the treatment of Dravet syndrome seizures.This article mainly reviews the research progress of unique clinical phenotype, SCN1A gene mutation characteristics and new antiepileptic drugs of Dravet syndrome, in order to deepen clinicians′ understanding of the disease.

Abnormal expression and dysfunction of voltage-gated Calcium channels (VGCCs) can give rise to a variety of neurological disorders in children, including epilepsy, migraine and ataxia.In the past, only CACNA1A, CACNA1H, CACNA2D2 and CACNB4 were considered associated with epilepsy in children.In recent years, an increasing number of VGCCs gene associated with epilepsy in children have been found, especially developmental and epileptic encephalopathy genes.This study aims to review the research progress of VGCCs gene mutations associated with epilepsy in children.