Biliary atresia (BA), an inflammatory sclerosing cholangiopathy, is the leading cause of cholestasis in infants. Pathologic features of BA include progressive inflammation and intrahepatic and extrahepatic bile duct fibrosis. BA is charac?terized by rapid liver fibrosis. The activation of hepatic stellate cell (HSC) is most important in liver fibrosis. Many mecha?nisms are involved in this process. miRNA can promote the activation of HSC through a variety of signaling pathways by regu?lating the expression of target gene, then playing a regulatory role in the synthesis and degradation of extracellular matrix (ECM). A lot of literatures show that PI3K/Akt is closely related to the occurrence and development of hepatic fibrosis. PI3K/Akt signaling pathway is involved in the activation of HSC proliferation and apoptosis. MiRNA activates PI 3K/Akt signaling pathway through various target genes, and then activates HSC to promote the development of liver fibrosis. In this paper, the miRNA related to biliary atresia of liver fibrosis is summarized.

Cholangitis is one of the most common complications after Kasai operation in children with biliary atresia (BA), whose precise etiology is still unclear. The occurrence of cholangitis may be the results of concurrent effects of various factors such as the structural change of intrahepatic bile duct or insufficient volume of bile flow. The diagnosis for cholangitis is based on clinical manifestations nowadays, and combined therapies including antibiotics, steroids and hepatoprotectants have been used empirically. The prophylaxis and treatment of postoperative cholangitis is the key to improve the liver survival of biliary atresai patients. Only by figuring out the etiology of cholangitis that we can prevent it and make sure the long-term survival of BA patients.

Objective To screen the high-efficiency predominant bacteria which can decompound 2-mercaptobenzothiazole, the accelerant for producing latex, in the organic wastewater. Methods Sampling from manufacturing environment, we got the predominant bacteria by primary screening, isolating and functional tests, and performed simulated test ground decompounding tests by using all bacteria. The enrichment of the predominant bacteria was followed by screen and identification to select the high-efficiency bacteria. Results 75 strains of predominant bacteria were obtained by primary screening. The simulated decompounding tests were performed after the mixed bacteria were tamed. The ratio of elimination for chemical oxygen demand (COD) was about 60.8%-97.7%, and the average was 77.2%. The predominant bacteria adhered to the surface of the active carbon, the carrier, and formed the biological film. Through screening and identification the Bacillus cereus showed to be predominant (90%). Conclusion The technology of high-efficiency predominant bacteria can be used for decompounding 2-mercaptobenzothiazole in the organic wastewater.

Objective:To investigate the effect of miRNA-1-3p (miR-1-3p) on expression of myocyte enhancer factor 2A (MEF2A) and the biological function of osteosarcoma cells.Methods:The tumor tissues and adjacent normal tissues of 20 patients with osteosarcoma who were clinically diagnosed in Shanxi Provincial Cancer Hospital from January 2019 to January 2020 were collected, and the expression of miR-1-3p in the samples was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The expression of miR-1-3p in osteosarcoma cell lines U2-OS, SAOS-2, MG63, SW1353 and human normal osteoblast cell line hFOB1.19 was detected by qRT-PCR, then the cell line with the lowest expression of miR-1-3p was selected for follow-up experiments. An overexpression miR-1-3p vector was constructed (miR-1-3p mimcs). The miR-1-3p overexpression group was transfected with miR-1-3p mimcs, and the control group was transfected with empty vector (miR-1-3p nc). CCK-8 method was used to detect the proliferation activity of cells; flow cytometry was used to detect the changes of cell apoptosis and cell cycle. miRwalk database was used to predict the miR-1-3p target gene, and the target gene was verified by dual-luciferase reporter gene assay; Western blot was used to detect the expression of MEF2A protein in cells of each group.Results:Compared with adjacent tissues, the expression of miR-1-3p in osteosarcoma tissues was down-regulated (0.31±0.14 vs. 0.62±0.21), and the difference was statistically significant ( t = 5.31, P<0.01). The expression of miR-1-3p in U2-OS cells was the lowest; compared with the control group, the proliferation activity of U2-OS cells was inhibited in miR-1-3p overexpression group (48 h absorbance value 0.56±0.01 vs. 0.77±0.03, t = 2.77, P<0.01; 72 h absorbance value 0.87±0.02 vs. 1.40±0.03, t = 2.93, P<0.01); G 1/S cell cycle arrest increased [G 1 phase (38.24±0.55)% vs. (32.11±0.80)%, t = 9.27, P = 0.01; S phase (61.24±0.90)% vs. (67.78±0.83)%, t = 7.52, P = 0.02]; early apoptotic rate increased [(11.20±0.12)% vs. (1.50±0.12)%, t = 2.91, P<0.05], miRwalk database predicted that the miR-1-3p target gene was MEF2A. The result of dual-luciferase reporter gene assay showed that miR-1-3p bound to MEF2A 3'UTR, and the luciferase activity of U2-OS cells in miR-1-3p overexpression group was lower than that in the control group (renilla luciferase/firefly luciferase activity ratio 0.53±0.06 vs. 1.00±0.04, t = 4.04, P < 0.05). Western blot showed that the expression of MEF2A protein in U2-OS cells of miR-1-3p overexpression group was lower than that of the control group (protein relative expression 0.41±0.14 vs. 0.77±0.12, t = 3.93, P < 0.05). Conclusions:The low expression of miR-1-3p may be associated with the proliferation, apoptosis and cycle changes of osteosarcoma cells. miR-1-3p can negatively regulate the expression of MEF2A protein and regulate the occurrence and development of osteosarcoma.

Objective To examine the association between the polymorphisms of brain-derived neurotrophic factor (BDNF)gene with heroin dependence.Methods Genomic DNA was isolated from the venous blood leukocytes of 308 unrelated patients with heroin dependence and 31 7 healthy individuals.Seven single nucleotide polymorphisms (SNPs)were genotyped using MassARRAY system.Data were analyzed using HaploView 4.0 and SPSS 20.0 software.Results There was a significant difference in the genotype frequency of rs6265 between heroin dependence group and healthy control group (χ2 =1 5.1 5 1,P =0.001).The rs6265 G allele was significantly higher than in controls (χ2 =9.864,P =0.002,OR =1.429,95% CI =1.143 -1.786).Furthermore,there was also a significant difference in the genotype frequency of rs13306221 between heroin dependence group and control group (χ2 =7.699,P =0.006).The rs13306221 G allele was significantly higher than in controls (χ2 =7.137,P =0.008,OR =0.539,95% CI =0.340-0.853).Strong linkage disequilibrium (LD)was observed in one block (D’> 0.9;r 2 >0.8),and significantly less G-G haplotype frequency of block 1 (χ2 =4.546;P =0.033)was found in heroin dependence group. Conclusion Our findings support the role of BDNF rs6265 and rs13306221 polymorphisms in heroin dependence and may guide future studies to identify other genetic risk factors for heroin dependence.

Ischemic stroke is an important cause of death and disability.Hemorrhagic transformation (HT) is a serious complication of acute ischemic stroke,especially in patients receiving intravenous thrombolytic therapy.Identifying patients at high risk of HT is very important for improving stroke outcomes.Some studied have shown that several serum biomarkers can be used to predict HT risk in patients with acute ischemic stroke.This article reviews the serum biomarkers of HT after acute ischemic stroke.

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

10.3969/j.issn.2095-4344.2013.25.019

To explore a new way of constructing bioartificial renal tubule assist device (RAD) in vitro and its function of transporting sodium (Na(+)) and glucose and to evaluate the application of atomic force microscope in the RAD construction, rat renal tubular epithelial cell line NRK-52E was cultured in vitro, seeded onto the outer surfaces of hollow fibers in a bioreactor, and then cultured for two weeks to construct RAD. Bioreactor hollow fibers without NRK-52E cells were used as control. The morphologies of attached cells were observed with scanning electron microscope, and the junctions of cells and polysulfone membrane were observed with atomic force microscope. Transportation of Na(+) and glucose was measured. Oubaine and phlorizin were used to inhibit the transporting property. The results showed that NRK-52E cells and polysulfone membrane were closely linked, as observed under atomic force microscope. After exposure to oubaine and phlorizin, transporting rates of Na(+) and glucose were decreased significantly in the RAD group as compared with that in the control group (P<0.01). Furthermore, when the inhibitors were removed, transportation of Na(+) and glucose was restored. It is concluded that a new RAD was constructed successfully in vitro, and it is able to selectively transport Na(+) and glucose.

Objective To investigate the expression and significance of integrin αvβ8, p38 and extracellular signal-regulated protein kinase 1/2 (ERK1/2) proteins, which are TGF-β1 pathway related regulatory protein, in liver fibrosis of children with biliary atresia (BA). Methods Fifteen cases of BA (Kasai group) and 10 cases of congenital biliary dilatation (CBD group) were collected in Tianjin Children’s Hospital. And liver biopsy specimens were collected in Tianjin first central hospital, including 10 cases of BA children who underwent liver transplantation due to liver failure after Kasai operation (liver transplantation group). The specimens of front part of the right lobe of the liver were taken for HE and immunohistochemistry (IHC) staining. The expressions ofαvβ8, p38 and ERK1/2 in liver were observed by IHC staining in three groups of liver tissues. Results HE staining showed fibroblast hyperplasia occasionally in CBD group, portal area expansion, fibrous tissue proliferation and wide spread bridging fibrosis with few pseudo lobules in Kasai group. In transplantation group, portal area was widened, the degree of fibrosis was severe and bridging fibrosis generally formed resulted in pseudo lobules widely. Imunohistochemistry showed that the expressions of αvβ8 and ERK1/2 were weakly positive, and the expression of p38 was negative in CBD group. In Kasai group, the expressions of αvβ8, p38 and ERK1/2 proteins were all strongly positive in liver cytoplasm, biliary epithelial cells and vascular endothelial cell cytoplasm. In liver transplantation group the expressions of αvβ8, p38 and ERK1/2 proteins were all strongly positive. The semi-quantitative analysis showed that the expressions levels of αvβ8, p38 and ERK1/2 were significantly higher in Kasai and liver transplantation groups than those of CBD group (P<0.05). There were no significant differences in expression levels ofαvβ8, p38 and ERK1/2 between Kasai group and transplantation group (P>0.05). Conclusion The expressions ofαvβ8, p38 and ERK1/2 are gradually increased in liver of BA with the process of fibrosis, which indicate that they may be involved in
the process of BA liver fibrosis.