BACKGROUND:Polysulfone membrane holds good anti-biodegradation ability, but how to use it to prepare hol ow fiber dialysis membrane and its blood compatibility have not been ful y understood. OBJECTIVE:To study the preparation, transfer property and biocompatibility of hol ow fiber dialysis membrane. METHODS:With polysulfone as the film material, diethylene glycol as the porogen, polyvinyl pyrrolidone as the modifier, N, N-dimethylacetamide as the solvent, and the hol ow fiber dialysis membrane was prepared using nonsolvent-induced phase separation. The performance was measured using scanning electron microscopy, ultra-depth three-dimensional microscope imaging and porosity test;the transfer parameters including reject rate and water flux were detected by ultrafiltration device;the blood compatibility was determined through hemolysis test, dynamic clotting time test and platelet adhesion test. Type II medical polyurethane material served as negative control. RESULTS AND CONCLUSION:The section of hol ow fiber dialysis membrane was asymmetric. 17%dialysis membrane showed a porous middle layer, while 19%, 21%and 23%membrane showed a sponge-like middle layer. Under the same membrane area, the density of fiber dialysis membrane was significantly lower than that of the negative control material, and the porosity of fiber dialysis membrane was significantly higher than that of the negative control material (P<0.05). The water volume and water flux of the hol ow fiber dialysis membrane were significantly higher than those of the negative control material (P<0.05). Results from three hemolytic tests showed that the average absorbance values and hemolysis rate of the hol ow fiber dialysis membrane were significantly higher than those of the negative control material (P<0.05). The dynamic clotting time test and the platelet adhesion test revealed that the dynamic clotting time of hol ow fiber dialysis membrane at 20, 40 and 70 minutes was significantly shorter than that of the negative control material (P<0.05). These results suggest that polysiloxane can be used as the membrane material to prepare hol ow fiber dialysis membrane using nonsolvent-induced phase separation, and holds a good biocompatibility, blood compatibility and transfer efficiency.

Objective To explore the clinical features of very long chain acyl-CoA dehydrogenase deifciency (VLCADD). Methods The clinical manifestation and the biochemical data of one baby girl with VLCADD were summarized and related literatures were reviewed. Results Female patient, aged 7 months, presented with recurrent vomit, haematemesis, stare, nodal tachycardia, hypoglycemia, abnormal liver function and myocardial enzyme. She died after one month due to frequent relapse together with withdrawing treatment. Conclusion VLCADD as a rare disease that cause sudden unexpected death in infant, the early diagnosis and treatment are important to patients.

Objective To explore the optimal condition for the differentiation of rat bone marrow mesenchymal stem cells(MSCs) into cardiomyocyte-like cells in vitro by using DMSO.Methods SD rat MSCs were isolated from rat bone marrow and cultured,then induced by 1.0% DMSO for 24 h.The cultured cells were observed through phase-contrast microscopy.Immunohistochemical technique and laser scanning confocal microscopy(LSCM) were used for detecting the expression of desmin,?-sarcomeric actin and C-TnT.The ultrastructures were examined with transmission electron microscopy.Results MSCs induced by DMSO were identified by the positive staining for desmin,?-sarcomeric actin and C-TnT.Transmission electron microscopy showed that paralleled myofilaments and a lot of rough endoplasmic reticulum were formed.Conclusion MSCs can be induced to differentiate into cardiomyocyte-like cells by DMSO.

This article was aimed to study An-shen drug substances of semen ziziphi spinosae and the relationship between index component distribution in vivo and meridian tropism. Intragastric administration of thyroid tablet suspension at the dose of 160 mg·kg-1 was given for 13 days for the establishment of yin deficiency rat model. Elevated plus maze test (EPM) was combined with light/dark box test to evaluate the effect of semen ziziphi spinosae on anxiety behavior among yin deficiency rats. The yin deficiency anxiety model rats' eyeballs were picked for blood at 10, 20, 30, 40, 60, 90, 120, 240 min after intragastric administration of semen ziziphi spinosae decoction. The rats were sacrificed for the collecting of heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine and small intestine and other tissues. HPLC-PDA-ELSD was combined to detect concentrations of spinosin, jujuboside A and jujuboside B in different tissues of rats. Related pharmacokinetic parameters were achieved after processing detected data with DAS 2.0 software. The results showed that compared with the yin deficiency group, semen ziziphi spinosae significantly reduced the rats' abnormal increased food-intake (P < 0.01) and water intake (P < 0.01) within 24 hours; significantly increased the body weight difference before and after treatment (P < 0.01); significantly reduced the kidney coefficient (P < 0.01) and adrenal gland coefficient (P < 0.01); significantly reduced the value of T3 (P < 0.01) and T4 (P < 0.05); significantly increased the value of TSH (P < 0.01). It showed that semen ziziphi spinosae can obviously improve yin deficiency symptoms. It significantly increased the number of open arms entering percentage (P < 0.01), the open arms holding percentage (P < 0.01), and the box through times in light/dark box test (P < 0.01). It showed obvious anti-anxiety effects. The index component distribution in vivo results showed that spinosin and jujuboside A were widely distributed in the heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine, small intestine and other tissues through blood circulation. Jujuboside B was distributed in the blood, stomach, large intestine and small intestine through blood circulation. The order of average concentration of spinosin among tissues was small intestine > stomach > liver > brain > large intestine > spleen > lungs > heart > kidneys. The order of AUC0-t in tissues was small intestine > stomach > liver > large intestine > spleen > brain > heart > kidneys > lungs. The order of average concentration of jujuboside A among tissues was lungs > large intestine > heart > spleen > liver > kidneys > small intestine > stomach > brain. The order of AUC0-t in tissues was lungs > spleen > liver > heart > large intestine > brain > stomach > kidneys > small intestine. The order of average concentration of jujuboside B among tissues was large intestine > small intestine > stomach. The order of AUC0-t in tissues was large intestine > small intestine > stomach. It was concluded that semen ziziphi spinosae can obviously improve yin deficiency symptoms with good anti-anxiety effects. Spinosin and jujuboside A in semen ziziphi spinosae were the drug substances of An-shen effect. They were also the material basis of sweet and sour taste. The spinosin and jujuboside A distribution in vivo of yin deficiency anxiety model rats were close to the meridian tropism of semen ziziphi spinosae.

Objective To identify the rale of NKX2-5 gene in cardiomyocyte differentiation and its mechanism.Methods P19 cells were divided into transfected and non-transfected groups.In the transfected group,P19 cells were with stable expression of NKX2-5 gene.The P19 cells were cultured in suspension for 4 days,and the formed aggregates were transferred to Petri dish for adherent culture.On days 4,8,12,and 16 of the adherent culture,the expressions of ct-saicomeric actin(α-SA)and cardiac troponin T(cTnT)were detected with double-labeling immunofluorescence and Western blot.The ultrastruetural changes were observed on day 16.Results In the transfected group,no expression of α-SA and cTnT was found on day 4,and the expression of these 2 proteins or co-expression existed on days 8,12,and 16.There were early cell junction and myofilament-like structure in the cytoplasm of some cells in the transfected group.In the non-transfected group,these 2 proteins were negative,and no differentiated cell was found.Conclusion Stable expression of NKX2-5 gene can induce cardiomyocyte differentiation from P19 cells,but the P19 cells with stable expression of JVKX2-5 gene is not suitable to be an in vitro model of cardiac development.

Purpose To study the clinicopathological characteristics of primary pancreatic neuroendocrine neoplasms. Method 60 cases of resected pancreatic neuroendocrine neoplasms according to the WHO (2010) classification of the digestive system of neuroen-docrine tumor to evaluate morphological standard, and combining with the literature to discuss the clinicopathological characteristics. Results Among the 60 cases, 23 cases were male patients, the rest were females, with male and female ratio of 1 ∶ 1. 61. The age of the patients were ranged from 19 to 69 years, with mean age of 49. 38 ± 11. 60 years. Tumor maximum diameter ranged from 0. 5 to 16 cm, and the mean diameter was 3. 29 ± 3. 53 cm. 30 cases located in the pancreatic head, 27 cases in the body and end of the pancre-as and 3 cases in the neck. Pathological examination showed the G1 (24 cases), G2 (25 cases), G3 (9 cases), and mixed adenon-euroendocrine carcinoma ( MANEC) in 2 cases. Immunohistochemical staining showed that NSE, CgA, Syn, and CD56 were diffusely positive expression. 45 patients were followed up for 4~80 months, 7 cases died, of which 1 case was G2, 4 cases were G3, and 2 ca-ses were MANEC. Conclusion Primary pancreatic neuroendocrine neoplasms is a relatively rare pancreatic malignant tumor, and the diagnosis is based primarily on histologic features and immunohistochemical examination. Accurate pathological assessment has impor-tant value to guide clinical treatment and prognosis.

Objective: o investigate the features of pathogenic bacteria for community-acquired bloodstream infection due to Gram-negative bacilli in patients with liver cirrhosis and optimal therapeutic strategy. Methods: A retrospective analysis was performed for the clinical data of patients with liver cirrhosis who were admitted to 302 Hospital of PLA due to community-acquired bloodstream infection from January 2010 to December 2015, and a statistical analysis was performed for their clinical features, pathogenic bacteria, and results of drug sensitivity test. The Pearson chi-square test was used for comparison of rates, and the Wilcoxon rank sum test was used for comparison of ranked data. Results: A total of 240 patients (including 178 male patients) with liver cirrhosis caused by various reasons were enrolled, with a mean age of 51.7 ± 11.1 years, an overall clinical remission rate of 80.42%, and an ineffective/mortality rate of 19.58%. The patients who used sensitive antibiotics within 12 hours after the onset of community-acquired bloodstream infection achieved a significantly higher improvement rate than those who used such drugs at more than 12 hours after onset (88.2% vs 58.1%, P < 0.001). The improvement rate achieved by the application of sensitive antibiotics at more than 12 hours after onset decreased with the increase in the Child-Pugh grade (P < 0.05). A total of 245 strains of Gram-negative bacilli were isolated, among which the six most common ones were 135 strains of Escherichia coli (55.1%), 62 strains of Klebsiella pneumoniae (25.3%), 16 strains of Aeromonas (6.5%), 4 strains of non-typhoidal Salmonella (1.6%), 3 strains of Enterobacter cloacae (1.2%), and 2 strains of Acinetobacter baumannii (0.8%). These Gram-negative bacilli had the highest sensitivity to meropenem (98.5%), followed by imipenem (97.9%), amikacin (97.5%), piperacillin/tazobactam (94.7%), cefmetazole (93.7%), and cefoperazone/sulbactam (93%). Different bacteria had different sensitivities to antibiotics. Conclusion Once community-acquired bloodstream infection occurs in patients with liver cirrhosis, highly sensitive antibiotics should be used as early as possible. Cefoperazone/sulbactam, piperacillin/tazobactam, imipenem, and meropenem can be used as first-line empirical antibiotics, and drug combination should be considered when necessary.

Heparinase III is an enzyme that specifically cleaves certain sequences of heparan sulfate. Previous reports showed that this enzyme expressed in Escherichia coli was highly prone to aggregation in inclusion bodies and lacks detectable biological activity. In this paper, we fused a glutathione-S-transferase (GST) tag to the N-terminus of heparinase III gene and expressed the fusion protein in Escherichia coli to develop an expression system of soluble heparinase III. As a result, approximately 80% of the fusion protein was soluble. The protein was then purified to near homogeneity via one-step affinity chromatography. A 199.4-fold purification was achieved and the purified enzyme had a specific activity of 101.7 IU/mg protein. This represented 32.3% recovery of the total activity of recombinant GST-heparinase III. The maximum enzyme production was achieved when bacteria were induced with 0.5 mmol/L isopropyl-beta-D-thiogalactoside at 15 degrees C for 12 h. The enzyme showed maximum activity at 30 degrees C and pH 7.5. And the enzyme activity was stimulated by 1 mmol/L Ca2+ and 150 mmol/L NaCl.
Escherichia coli ; genetics ; metabolism ; Flavobacterium ; enzymology ; genetics ; growth & development ; Glutathione Transferase ; biosynthesis ; genetics ; Heparin Lyase ; biosynthesis ; genetics ; isolation & purification ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Objective To develop a tetra-primer amplification refractory mutation system PCR(T-ARMS-PCR)assay for detecting four single nucleotide polymorphisms(SNPs): rs1801133,rs1801131, rs1805087 and rs1801394 associated with folate metabolism in a single reaction tube.Methods Methodology was developed.Then, it applied the established method to analyze 150 physical examination children′s anticoagulant blood samples admitted to the Department of Clinical Laboratory, Children′s Hospital of Hebei Province from January 2017 to April 2017.Four sets of chimeric primers consisting of a universal Tag sequence and targeting rs 1801133,rs1801131,rs1805087 and rs1801394 fused to the specific sequence were designed according to the T-ARMS-PCR principle and the multiplex chimeric primers strategy.A single tube PCR was then conducted after optimizing the primer concentrations and the reaction conditions.The amplified products were analyzed by QIAxcel capillary electrophoresis, and the corresponding SNP genotypes of 150 samples were identified.Furthermore,all samples were verified by direct sequencing.And the Hardy-Weinberg Equilibrium(HWE)testing of four SNPs from 150 samples were conducted by SPSS17.0 Chi-square test.Results The improved T-ARMS-PCR combined with capillary electrophoresis can accurately verify eight different alleles of the four SNPs associated with folate metabolism at one time in 3 hours.Four SNPs of 150 whole blood samples were accurately classified and the results were completely consistent with direct sequencing.All the genotype frequencies of these four SNPs were in HWE (χ2rs1801133=0.69, Prs1801133=0.40; χ2rs1801131=0.21, Prs1801131=0.64; χ2rs1805087=3.32, Prs1805087=0.07;χ2rs1801394=1.91, Prs1801394=0.17).Conclusions The proposed improved T-ARMS-PCR combined with capillary electrophoresis in this study can accurately verify four SNPs associated with folate metabolism in a single reaction tube.This method might be a valuable tool to specifically guide the folate supplement in general population.

Objective To explore whether the norepinephrine transporter(NET) gene T-182C and G1287A polymorphisms are involved in the etiology of schizophrenics among the Chinese Han population.Methods 249 schizophrenics(case group) and 309 healthy physical examiners(control group) of the Chinese Han population were collected.Ligase detection reaction was used to detect the genotype distributions and allele frequencies of NET rs2242446 and rs5569 polymorphisms in the two groups.The genotype distributions and allele frequencies of these two single nucleotide polymorphisms (SNPs) were also compared in the case and control groups.Results In the case group,the TF genotype of the NET gene loci rs2242446 was the most,48.6％,and the CC genotype of which was the least,8.4％.In the control group,the CT genotype was the most,47.9％,and the C C genotype was the least,11.3％,the minimum allele frequency was 29.9％.In the case group,the GG genotype of the NET gene loci rs5569 was the most,46.2％,and the CC genotype was the least,9.6％.In the control group,GG genotype was the most,51.5％,and the AA genotype was the least,10.3％,and the minimum allele frequency was 29.4％.No significant differences in the genotype and allele distribution of NET rs2242446 and rs5569 were found in Chinese-Han patients with schizophrenia and control participants (all P＞ 0.05).In gender-specific analyses,similarly,no significant differences were found for rs2242446 and rs5569 genotype and allele distributions in either the male or the female case-control comparisons (all P＞ 0.05).Conclusion The NET rs2242446 and rs5569 polymorphisms are not associated with schizophrenia in Chinese Han population.