Objective: To study the probiotic properties of 12 selected strains of lactobacilli.Methods: The probiotic properties of 12 selected strains of lactobacilli were investigated for their resistance to pH 2.5 and 0.3% oxgall,their adhesion to Caco-2 cells and antimicrobial activity against enteric pathogenic bacteria.Results: The 12 strains all showed relatively high resistance to acid and bile salt and varied antimicrobial activities against pathogenic bacteria of intestinal infection.Lactobacillus johnsonii JCM1022 exhibited a broad inhibitory effect on enteric pathogenic bacteria,especially on enteropathgenic E.coli and Salmonella enteritidis,while Lactobacillus reuteri JCM1081 possessed a high activity of adhesion to Caco-2 cells(495.07?80.03).Conclusion: Among the 12 strains,Lactobacillus johnsonii JCM1022 and reuteri JCM1081 are qualified candidates for clinical application.

Objective The goal of the study was to assess the lethal effect of cytotoxic lymphocyte against A549 cells induced by dendritic cells (DC) pulsed with A549 lysate and transfected with GM-CSF recombinant adenovirus. Methods The cytotoxic lymphocyte against A549 cells were induced by culturing with DCs, which pulsed with A549 antigens and transfected with GM-CSF recombinant adenovirus. The of effector/target ratio, the killing rates of N-DC group, A-DC group and G-A-DC group were (1.9±0.7) %,effector/target ratio, the killing rates of N-DC group, A-DC group and G-A-DC group were (5.3±0.2) %, (40.5±7.7) % and (72.5±4.7) %, respectively. We found that the killing rate of G-A-DC group was the highest by statistics. Conclusion The cytotoxic lymphocyte against A549 cells can be induced by DCs pulsed with A549 lysate ,and the lethal effect of CTLs can be enhanced when DCs were infected with GM-CSF recombinant adenovirus.

Objective To investigate the expression of CD24 and FAK in the invasive breast ductal carcinoma tissues and their clin-ical significance. Methods Tissues were obtained from 86 patients with breast carcinoma and 30 non-cancer specimens. The expression of CD24 and FAK was detected by immunohistochemical technique (ElivisionTMPlus) and their correlation with tumor clinicopathological char-acters was analyzed. Results The positive rate of CD24 and FAK expression in 86 cases of breast carcinoma was 87.2% (75/86) and 72.1% (62/86) respectively, which was significantly different from that in control group(P<0.01). Expression of CD24 was significantly correlated with axillary lymph node metastasis(P<0.01). Expression of FAK was correlated with histopathologic grade, axillary lymph node metastasis and clinical stage (P<0.05). There was a positive correlation between the expressions of CD24 and FAK in breast carci-noma tissues (rs = 0.222, P < 0.05). Conclusion Expression of CD24 and FAK are closely correlated with the development of breast carcinoma and there is a markedly positive correlation between CD24 and FAK expression. These indexes can be helpful in understanding the biological behaviors and predicting the prognosis of breast carcinoma.

To determine the effect of pre-operative chemotherapy on apoptosis in breast cancer and to evaluate its signif icance as a prcgnostic marker. MethodsPatients with breast cancer were divided into preoperative chemotherapy group (40 cases)and control group (42 cases). Two groups were analyzed for the appearance of apoptosis by using TUNEL method and electron mi croscope in tissue sections. ResultsApoptosis occurred in 92.5 % of preoperative chemotherapy group and in 78.5 % of control group. The apoptotic indexes were 19.37 + 6.49 and 9.26 + 5.04 ( P ＜ 0.01 ) respectively. Low apoptotic index was related to disease-free survival of patients with breast cancer (P ＜ 0.01 ). ConclusionThe preoperative chemotherapy can induce apoptcsis of breast cancer and improve disease-free survival.

Objective To investigate the expression of CD24 and Ki67 in the invasive breast ductal car-cinoma tissues, so as to assess the role of CD24 protein in the carcinogenesis and progression of breast canc-er. Methods The expression of CD24 and Ki67, Bcl-2 was examined by immunohistochemically in 86 breast cancer specimens and 30 non-cancer specimens ( ElivisionTM Plus). The relationship between CD24 expression and Ki67, Bcl-2, patient age, tumor size, histopathologic grade, axillary lymphnode metastasis, and clinical stage of breast cancer was analyzed. Results In 86 eases of invasive breast ductal carcinoma, the positive incidences of CD24 was 87.2% (75/86), which distinguished apparently from those of the con-trol group( P <0.01 ). In breast carcinoma tissues, the expression of CD24 was positively related to axillary lymphnode metastasis(P <0.01 ), but was not related to patients age,tumor size, histopathologic grade and clinical stage(P>0.05). CD24 expression was positively related to Ki67 (P<0.01 ) and negatively relat-ed to Bcl-2(P>0.05). Conclusion CD24 expression was related to the proliferation, invasion and me-tastasis of breast cancer cells and it can serve as an important marker for predicting biological behavior and prognosis of tumors.

OBJECTIVETo explore the molecular mechanism for a boy suspected with 3-methylcrotonyl-CoA carboxylase deficiency by neonatal screening.

METHODSPCR and Sanger sequencing were used to identify potential mutations of MCCC1 and MCCC2 genes. SIFT and Polyphen-2 software was used to predict the effect of variant on the protein function and conservation of the variant across various species. Human Splicing Finder and Swiss-PdbViewer4.1.0 were applied to analyze the possible mechanism of the variant.

RESULTSFor the proband, a compound heterozygous mutation was discovered in the MCCC1 gene, namely c.539G>T (p.G180V) and c.704_711del (p.A235Vfs*4), which were inherited from his father and mother, respectively. The two mutations have disrupted the protein conformation, which in turn may impact the function of MCC protein.

CONCLUSIONThe compound heterozygous mutations of the MCCC1 gene may contribute to the 3-methylcrotonyl-CoA carboxylase deficiency manifested by the patient.

Amino Acid Sequence ; Base Sequence ; Carbon-Carbon Ligases ; chemistry ; deficiency ; genetics ; DNA Mutational Analysis ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Models, Molecular ; Mutation ; Neonatal Screening ; methods ; Protein Conformation ; Sequence Homology, Amino Acid ; Urea Cycle Disorders, Inborn ; diagnosis ; genetics

OBJECTIVETo explore the molecular etiology for a Chinese family affected with isolated methylmalonic acidemia (MMA).

METHODSPotential mutations of MUT, MMAA and MMAB genes in the proband were screened by PCR and Sanger sequencing. The pathogenicity of identified mutations was analyzed using Polyphen2, SIFT, HSF, DNAMAN 6.0 and Swiss-PdbViewer4.1.0 software.

RESULTSTwo novel mutations of the MUT gene, including c.581C>T (p.P194L) and c.1219A>T (p.N407Y), were discovered in the proband, which were inherited respectively from his mother and father. Bioinformatics analysis suggested that both mutations were damaging. The affected codons P194 and N407, both located in the (beta, alpha) 8 barrel domain and to which the substrate methylmalonyl-CoA is bound, are highly conserved across various species. Both mutations can disrupt the space conformation of its protein product, affecting the function of the MCM protein.

CONCLUSIONThe novel mutations of MUT gene probably underlie the isolated MMA in this family.

Adult ; Amino Acid Metabolism, Inborn Errors ; enzymology ; genetics ; Amino Acid Sequence ; Animals ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Female ; Humans ; Infant ; Male ; Methylmalonyl-CoA Mutase ; genetics ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; Pedigree ; Point Mutation ; Sequence Alignment

OBJECTIVETo explore the value of single nucleotide polymorphism array (SNP-array) for the analysis of pediatric patients with growth retardation.

METHODSOne hundred eighty one children with growth retardation were enrolled. DNA was extracted from peripheral samples from the patients, and whole genome copy number variations (CNVs) were detected using Illumina Human Cyto SNP-12. All identified CNVs were further analyzed with reference to databases including ClinGen, ClinVar, DECIPHER, OMIM and DGV as well as comprehensive review of literature from PubMed to determine their pathogenicity.

RESULTSForty seven patients (26%) with abnormal CNVs were detected, which included 12 known microdeletions/microduplications syndrome (26%), 10 pathogenic non-syndromic CNVs (21%), 3 numerical chromosome aberrations (6%), 3 unbalanced translocations (6%), 4 pathogenic mosaicisms (9%) and 15 cases with unknown clinical significance (32%). After excluding obvious numerical and/or structural chromosomal abnormalities, this study has detected 15 pathogenic microdeletions/microduplications sized 5 Mb or less, which may be missed by routine chromosomal karyotyping. In addition, there were 3 cases with loss of heterozygoisty (LOH) containing known or predicted imprinting genes as well as 2 cases with suspected parental consanguinity.

CONCLUSIONSNP-array technology is a powerful tool for the genetic diagnosis of children with growth disorders with advantages of high resolution and improved accuracy.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; DNA Copy Number Variations ; Developmental Disabilities ; diagnosis ; genetics ; Female ; Humans ; Infant ; Karyotyping ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Polymorphism, Single Nucleotide

OBJECTIVETo identify the genetic mutation in ASS1 gene in a Chinese family with citrullinemia typeI, which may provide a basis for the diagnosis and genetic counseling.

METHODGenomic DNA was isolated from peripheral blood samples of the family members. Mutation analysis of ASS1 gene was carried out by PCR and Sanger sequencing. Biostructural analysis of the mutated ASS1 was completed by Phyre server.

RESULTDouble heterozygous mutations in the proband were identified: c.951delT (F317LfsX375) and c.1087C>T (R363W), which were confirmed in the proband's father and mother, respectively. It was found that the c.951delT mutation might change the formation of a dimer or a tetramer and the function of ASS1 protein.

CONCLUSIONDouble heterozygous mutations for c.951delT and c.1087C>T have been found in a proband with citrullinemia typeI. The c.951delT is a novel mutation in citrullinemia typeI, which may change the configuration of ASS1 protein and result in ASS1 dysfunction.

Argininosuccinate Synthase ; genetics ; Asian Continental Ancestry Group ; genetics ; Citrullinemia ; genetics ; DNA Mutational Analysis ; Humans ; Infant, Newborn ; Mutation

The emission of hydrogen sulfide in the waste gas from slaughter plant, fishmeal feed processing and some other food industrial processing could cause serious air pollution to the surrounding environment. The purpose of this study was to screen heterotrophic bacterium strains for the removal of hydrogen sulfide odor. One heterotrophic bacterial mutant ZJNB-B3 was derived from the sulfide degrader Bacillus cereus XJ-2 and its sulfide removal efficiency was 97%. Based on the morphology studies, biochemical tests and 16S rRNA gene analysis, the strain was identified as Bacillus cereus ZJNB-B3. The NCBI GenBank accession number is MF679650. Batch tests showed that the strain tolerated up to 300 mg/L of toxic S²⁻ concentration. Response surface methodology was applied to optimize the conditions of degradation of sulfide. The optimal parameters were as follows: initial sulfide concentration 211.8 mg/L, initial pH 6.72, inoculum volume 5.04%, and incubation temperature 30 ℃. The accumulated sulfate concentration was 63.8 mg/L and the sulfide removal efficiency was 97.3% after 48 h incubation. No sulfuric acid was generated during sulfide oxidation by the strain. Sulfide could be removed effectively by this strain under mild pH conditions. The results suggested that the strain may have great industrial application potential. This study provides the fundamentals for the removal of hydrogen sulfide gas.