Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Objective To evaluate the reliability and validity of hepatocellular carcinoma (HCC)using the liver imaging reporting and data system (LI-RADS).Methods By using the LI-RADS v2013.1,two radiologists evaluated 1 62 patients with cirrhosis or with a risk of HCC according to an inclusion criteria.The Kappa value was used to evaluate the consistency between two different diagnoses and was compared with pathological and follow-up results.The accuracy of the LI-RADS was assessed including sensitivi-ty,specificity,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio and accuracy rating.Results In all 1 62 patients,there were HCCs in 97 patients including 7 patients belonging to the LR3,benign diseases in 50,and other kinds of malignancy in 1 5.The Kappa value was 0.882 (P =0.000)between two observers on LI-RADS grading. The sensitivity,specificity and accuracy rating of LI-RADS grading in diagnosing HCC was 100.00%,91.30%,and 97.06%,re-spectively.Conclusion The LI-RADS has high consistency and stability in evaluation and diagnosis of HCC by enhanced CT.LR3 di-agnosis should be cautious because of a susceptible development to HCC,which can be improved through the combination of clinic and laboratory examination.

Objective To investigate the effect and mechanism of boron neutron capture therapy(BNCT) in C6 glioma cell line.Methods C6 cells in exponential phase were divided into 6 groups: untreated control,(~(60)Co?)(4 Gy),~(60)Co? 8 Gy,nuclear reactor exposure without boronophenylalanine(BPA) 3 Gy,BNCT(4 Gy) and BNCT 8 Gy.Cellular morphological change was observed by an inverted microscope,light microscope,fluorescence microscope and electronic microscope.Flow cytometry was used to determine the percentage of apoptosis,necrosis and normal cells 48h after irradiation.Colony forming assay was used to calculate cell surviving fraction.Results Typical morphological changes of apoptosis were observed early after irradiation in BNCT group,with a significant increase in apoptotic rates was observed 48 h after irradiation with 63.2% and 88.3% for BNCT(4 Gy) and 8 Gy group,respectively(P

The domestic pharmaceutical management profession started relatively late, and professionals in this field are urgently needed. It is especially important to cultivate and reserve pharmaceutical management professionals in traditional Chinese medicine (TCM) industry. Based on the background of pharmaceutical management discipline combed with the characteristics of TCM, this paper takes Beijing University of Traditional Chinese Medicine as an example to further explore the quality of pharmaceutical management professionals in TCM institutions. It is suggested that in the education of pharmaceutical management students in TCM colleges, students should understand and inherit our TCM culture, focus on philosophical thinking, cultivate their people-oriented management ability combined with Golden Mean, master the knowledge of TCM theory and general management from characteristic professional curriculums, and possess comprehensive qualities that are able to inherit ancient TCM culture with innovation.

Objective:To investigate the expressions of transforming growth factor-β1 (TGF-β1), aquaporin (AQP) and osteopontin (OPN) in renal injury induced by fluorosis in rats.Methods:According to body weight (80 - 100 g), forty-eight SD rats were divided into control group (normal saline), low fluoride group (10 mg/kg) and high fluoride group (20 mg/kg) by random number table, 16 rats in each group (half males and half females). The rats were exposed to fluoride by intraperitoneal injection of sodium fluoride. After the rats were treated with fluoride for 12 weeks, the rats were killed by femoral artery bloodletting and the renal tissue was taken. The contents of TGF-β1, AQP and OPN in serum of rats with fluorosis were analyzed by enzyme-linked immunosorbent assay (ELISA). The expressions of TGF-β1, AQP and OPN in renal tissue of rats with fluorosis were analyzed by quantitative real-time PCR (qRT-PCR) and immunocytochemistry.Results:The serum levels of TGF-β1, AQP and OPN in high fluoride group [(26.42 ± 6.09), (378.60 ± 36.84) μg/L, (603.45 ± 64.32) pg/ml] were higher than those in control group [(2.41 ± 0.42), (157.41 ± 15.26) μg/L, (182.45 ± 30.63) pg/ml] and low fluoride group [(13.15 ± 3.26), (245.65 ± 23.21) μg/L, (359.47 ± 55.26) pg/ml, P < 0.05]. The levels of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). The expressions of TGF-β1, AQP and OPN mRNA in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the expressions of TGF-β1, AQP and OPN mRNA in low fluoride group were higher than those in control group ( P < 0.05). The positive expression scores of TGF-β1, AQP and OPN in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the positive expression scores of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). Conclusion:TGF-β1, AQP and OPN are highly expressed in the renal tissue of rats with fluorosis, which can be used as indicators to judge the damage of renal aggregation system caused by fluorosis.

Objective To construct the LyP-1 targeted MR fluorescence dual-modality molecular probe for pancreatic cancer,and to observe its features and MRI charicteristics.Methods The 50 nm MR-fluorescent dual-modality molecular probe with surface modified with cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly Cys) was rationally designed.Whether the molecular probe could specifically recognize the pancreatic cancer cells were validated by the combination of fluorescent imaging and MR T2WI.Results The new MR-fluorescent dual-modality molecular probe anchored with LyP-1 could be used for the fluorescent imaging and MR T2WI of pancreatic cancer in mouse.And the molecular probe was demonstrated to be effective in conjugating with pancreatic cancer cells on fluorescent images and caused obvious MR signal reduction under T2 relaxometry in vitro.In vivo experiment,the molecular probe could be used for fluorescent labeling tumor tissue and detecting orthotopic pancreatic cancer in C57BL/6 mouse as MR contrast agent.Conclusion The LyP1 immobilized MR-fluorescent dual-modality molecular probe can actively target to mouse orthotopic xenograft of pancreatic cancer,which is hopeful to the application in early probing and diagnosis of pancreatic cancer by multimodal imaging.

Irritable bowel syndrome (IBS) is characterized by abdominal pain associated with changes in defecation frequency and blood folate level. FODMAP stands for fermentable oligosaccharides, disaccharides, monosaccharides and polyols. High-FODMAP diet could elicit or exacerbate IBS-associated bowel symptoms, which is inadequately recognized among gastroenterologists in China. Here we report an IBS case, focusing on the association between high-FODMAP diet and bowel symptoms and the intervention of low-FODMAP diet.

OBJECTIVE: To analyze the clinical phenotype and genetic basis for a child with acute form of tyrosinemia type I (TYRSN1). METHODS: A child with TYRSN1 who presented at the Gansu Provincial Maternal and Child Health Care Hospital in October 2020 was selected as the subject. The child was subjected to tandem mass spectrometry (MS-MS) and urine gas chromatography-mass spectrometry (GC-MS) for the detection of inherited metabolic disorders, in addition with whole exome sequencing (WES). Candidate variants were validated by Sanger sequencing. RESULTS: The child's clinical features included abdominal distension, hepatomegaly, anemia and tendency of bleeding. By mass spectrometry analysis, her serum and urine tyrosine and succinylacetone levels have both exceeded the normal ranges. WES and Sanger sequencing revealed that she has harbored c.1062+5G>A and c.943T>C (p.Cys315Arg) compound heterozygous variants of the FAH gene, which were inherited from her father and mother, respectively. Among these, the c.943T>C was unreported previously. CONCLUSION Considering her clinical phenotype and result of genetic testing, the child was diagnosed with TYRSN1 (acute type). The compound heterozygous variants of the FAH gene probably underlay the disease in this child. Above finding has further expanded the spectrum of FAH gene variants, and provided a basis for accurate treatment, genetic counseling and prenatal diagnosis for her family.
Female ; Humans ; Gas Chromatography-Mass Spectrometry ; Genetic Testing ; Mutation ; Phenotype ; Prenatal Diagnosis ; Tyrosinemias/genetics* ; Child

Based on previous bioinformational analysis results, two Aspergillus niger lipase (ANL) mutants, ANL-Ser84Gly and ANL-Asp99Pro were constructed to screen ANL mutants with oil-water interface independence. ANL-Ser84Gly still displayed a pronounced interfacial activation, while ANL-Asp99Pro displayed no interfacial activation. The specific activity of ANL-Ser84Gly towards p-nitrophenyl palmitate (-myristate, -laurate and -decanoate) decreased by 29.8% (53.1, 60.1 and 77.1, respectively) than that of ANL, while the specific activity of ANL-Asp99Pro towards p-nitrophenyl palmitate increased by 2.2-fold. The mutation in the hinge region at both sides of the lid domain also destabilized various secondary structure factors of ANL-S84G and ANL-D99P, which resulted in a substantial decrease in thermostability. The achievement to construct oil-water interface-independent ANL mutants would help to further understand lipase interfacial activation mechanism.
Aspergillus niger ; enzymology ; genetics ; Base Sequence ; Enzyme Stability ; Fungal Proteins ; genetics ; metabolism ; Lipase ; genetics ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Oils ; Substrate Specificity ; Water