miRNAs were discovered less than a decade ago, and have emerged as important regulators of gene expression in mammals. A large number of miRNAs have been identified to be located within the intronic regions of protein-encoding genes(host genes) and called intronic miRNAs. The intronic miRNAs may play a key role in regulating the expression and function of their host genes due to the fact that most of them are co-expressed with the host genes. In this paper, the recent advances on the research on potential relationship between intronic miRNAs and their host genes are reviewed.

Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.

OBJECTIVETo study the effect of red peony root (RPR) on serum proteome in rat suffering from noxious heat with blood stasis Syndrome (NH-BS).

METHODSThe differences of serum proteome among rats in four groups, treated with lipopolysaccharide (LPS), RPR, LPS + RPR and saline respectively, were analyzed by bi-dimensional electrophoresis (2DE) assay. LPS was administered by intravenous injection and RPR by oral intake.

RESULTS(1) Serum of rats with LPS induced NH-BS showed significant changes in volume of serum protein (xPr) in 13 points on 2DE collagen, the volume of xPr 16 and 19 were significantly lower, volume of xPr 1, 2, 3, 4, 6, 7, 8, 9, 11, 12 and 23 were significantly higher respectively, as compared with those in the normal control group. (2) After being treated with RPR, the increased volume of xPr 1, 2, 3, 4 and 9 significantly decreased, and the decreased xPr 16 significantly increased, with xPr 2, 3 restored to normal level but the xPr16 still lower and xPr 1, 4, 9 higher than those in the normal group. RPR showed interaction with LPS on xPr 1, 3, 9, and 16. (3) For xPr 19, the interaction of RPR with LPS might be synergistic. (4) In the group treated with RPR, volumes of xPr 13 and 14 were significantly higher and those of 15, 17 were significantly lower than those in the normal group respectively, but the similar changes didn't found in the LPS group.

CONCLUSIONThe molecular basis of therapeutic effect of RPR on NH-BS might be through the regulation of xPr 1, 2, 3, 4, 9 and 16.

Animals ; Blood Proteins ; metabolism ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Endotoxemia ; blood ; chemically induced ; drug therapy ; Lipopolysaccharides ; Male ; Medicine, Chinese Traditional ; Paeonia ; Phytotherapy ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley

?AlM: To investigate the outcome and safety of Ahmed glaucoma valve implantation treatment in uncontrolled primary congenital glaucoma ( PCG) .
? METHODS: Twenty - two eyes in 22 children with uncontrolled PCG were reviewed retrospectively and underwent Ahmed glaucoma valve implantation treatment from January 2011 to December 2014. Main checking index included intraocular pressure ( lOP ) before and after operation, corneal diameter and complications.
?RESULTS: Preoperative mean age was 3. 74±2. 24y, and 2. 59 ± 1. 78y apart from the last operation. Postoperative average lOP was 35. 22 ± 6. 36mmHg. Average corneal diameter was 12. 79 ± 0. 75mm. Mitomycin C ( 0. 3 - 0. 5mg/mL ) was used in all operations for 3-5min. Glaucoma valves were implanted in the temporal or nose above the equator sclera. Postoperative lOP was 11. 4±4. 45mmHg at 1wk, and 16. 73± 7. 23mmHg after 12mo. As lOP< 21mmHg for success criteria, lOP of 16 eyes ( 73%) were controlled after 12mo. Preoperative 6 cases had shallow anterior chamber, recovered spontaneously. No serious complication was recorded, such as rejection of glaucoma valve, endoophthalmitis and corneal decompensation.
?CONCLUSlON:Ahmed glaucoma valve implantation in uncontrolled PCG is a safe and viable treatment.

HEV is classified into H (human) group and Z (zoonosis) group according to its compatible host. H group contains genotype 1 and genotype 2 HEV isolates which infect human only; Z group contains genotype 3 and genotype 4 HEV isolates which infect both human and animals. After analysis of amino acid sequences between ORF2 aa368 and aa606, four group-conserved sites that were all located in the neutralization region of ORF2 were identified. They are aa483, aa492, aa497 and aa599. Mutation analysis and capture PCR were then performed on these sites with a group of monoclonal antibodies. Results showed that the difference of the aa497 between H and Z groups was responsible for the maintenance of their group-specific immunodominant epitopes, probably through confirmation-dependent epitope changes. Thus, aa497 and its related change on the surface structure of HEV may play important roles in host selection by H and Z groups of HEV.
Antibodies, Monoclonal ; immunology ; Base Sequence ; Genotype ; Hepatitis E virus ; classification ; genetics ; immunology ; Humans ; Immunodominant Epitopes ; Molecular Sequence Data ; Mutation ; Neutralization Tests ; Open Reading Frames

OBJECTIVETo investigate the changes in the expression level of sRNA SpR19 and its potential target protein GroEL in clinical isolates of Streptococcus mutans with different cariogenicity exposed to different pH conditions and explore the possibility of using these molecules as biomarkers for assessing the cariogenicity of the bacteria.

METHODSThe total RNAs were extracted from the clinical isolates of Streptococcus mutans with high (strain 17) and low cariogenicity (strain 5) for high-throughput sequencing for profiling of the differentially expressed sRNAs. The candidate sRNA, SpR19, was selected for further study on the basis of bioinformatics analysis considering the role of its potential target in the cariogenic process. The differential expression levels of SpR19 in the strains exposed to both pH5.5 and pH7 culture conditions were verified by quantitative real-time PCR. The expression of the potential target of SpR19, GroEL, was also investigated at both the protein and mRNA level using Western blotting and quantitative real-time PCR.

RESULTSBioinformatic analysis suggested multiple potential target sites of SpR19 both in GroEL mRNA and in the upstream and downstream inter-genic regions. Under different pH conditions, the highly cariogenic strain 17 expressed consistently low levels of SpR19 as compared with the strain 5 with a low cariogenicity; GroEL showed a reverse expression pattern in the 2 strains. An inverse correlation was found between the expressions of SpR19 and GroEL.

CONCLUSIONThe highly cariogenic strain 17 expressed low levels of SpR19 and high levels of GroEL in both acidic and neutral culture conditions. SpR19 may negatively regulate the cariogenicity of Streptococcus mutants by targeting at GroEL.

A fragment of hepatitis E virus open reading frame-2(ORF2), located from amino acid residues 394 to 604, was expressed in E. coli. The recombinant protein NE2 was found to form homodimer mostly in SDS-PAGE, which can be dissociated to monomers when treated with urea, and it was recognized more strongly in its dimeric form than the monomer by HEV reactive human serum in Western blotting. Besides, many aggregated form of NE2 from dimer to at least hexamer can be seen in MALDI-TOF-MS. And when the hydrated dynamic semidiameter of NE2 moleculars in PBS was measured as about 4 nm by Dynamic Light Scattering (DLS), being equal to tetramer, but with high polydispersity, which suggested that the NE2 moleculars were existed in PBS in many different sizes. These results suggested that the recombinant NE2 can aggregate into several oligomer forms, the association in the dimer is most strong, and dimers can assemble further to form some super-structure.
Dimerization ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Protein Conformation ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate the feasibility of 9Tc(m)-HL91 imaging in prediction of 34 radiotherapy sensitivity of naqsopharyngeal cancer( NPC) and its relationship with prognosis.

METHODSpatients with NPC confirmed by pathology, staging from II-IVa, underwent 99Tc(m)-HL91 SPECT imaging one week before radiotherapy. 18 of them received adjuvant chemotherapy. The hypoxia in primary nasopharyngeal lesions and cervical lymph node metastases were calculated semi-quantitatively, and compared with clinical findings in medium-term therapy at 4 months and 1 year post therapy.

RESULTS(1) There was no significant relationship between the total preliminary curative effect of adjuvant chemotherapy and the degree of nasopharyngeal lesion hypoxia (T/Mu, gamma = -0.394, P = 0.145; T/ Ce gamma = -0.510, P = 0.052). But there was a significant difference between the partial curative effect group and significant curative effect group. (2) The degree of NPC regression in the medium-term radiotherapy group was negatively correlated with the degree of hypoxia (T/Mu, gamma = -0.602; T/Ce, gamma = -0.643, P < 0.01). (3) 23 patients had good local control except one case with lung and bone metastasis 4 months post-therapy. The lesions disappeared or not developed in 6 patients (T/Mu 1.30 +/- 0.23, T/Ce 3.61 +/- 0.84). Two patients at stage III and IVa relapsed (T/Mu were 1.40 and 1.27, respectively; T/Ce were 4.10 and 3.85, respectively), there was no significant difference. (4) The degree of lymph node hypoxia had no correlation with the curative effect on medium-term radiotherapy.

CONCLUSION99 Tc(m)-HL91 hypoxia imaging may predict sensitivity to radiotherapy in patients with NPC, with a potential help to carry out individual therapy. However, further investigation is needed to ascertain whether it could predict the long-time curative effect on NPC radiotherapy.

Adult ; Aged ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Humans ; Hypoxia ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; drug therapy ; radiotherapy ; Neoplasm Staging ; Organotechnetium Compounds ; Oximes ; Particle Accelerators ; Preoperative Care ; methods ; Prognosis ; Radiotherapy, High-Energy ; Remission Induction ; Reproducibility of Results ; Tomography, Emission-Computed, Single-Photon

In this study, a new combined enzyme immunoassay(NRAg ELISA) for detection of HBV PreS1 and core antigens which was highly consistent with serum HBV DNA test was established. The serial serum dilution test indicated that the average sensitivity of the assay was 10(3.2) genome copies/mL (95% CI: 10(2.2-4.2) genome copies/mL), which was notably higher than the test performed on Pre S1 or core antigen alone. The test with sera from 994 blood donors whose HBsAg were negative demonstrated that the specificity of this assay was 99.7% (95% CI: 99.1%-99.9%). 271 serum samples from chronic hepatitis patients were also examined and the result showed that the total consistent rate between NRAg ELISA and HBV DNA was 96.3% (95% CI: 93.3%-98.2%). The NRAg ELISA S/CO(signal/cutoff) was closely correlated with HBV genome copies (R = 0.9158, n=231). Furthermore,by using this assay,we found a patient whose HBsAg was negative but HBV DNA was positive. Sequencing result showed that HBV genome from this patient had a point mutation in the "a"epitope of S gene. Our results indicate that HBV NRAg ELISA has a high relativity with HBV DNA test, and can effectively detect the mutation of HBsAg,it is expected to be a potent tool for screening HBsAg mutant and is a convenient method for substituting HBV DNA test.
DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Hepatitis B Antigens ; blood ; Hepatitis B Core Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results

OBJECTIVETo identify prognostic factors in patients with gastrointestinal stromal tumors (GIST).

METHODSHematoxylin and eosin (H&E) stained histopathological slides of tumors from patients with mesenchymal neoplasms growing in the gastrointestinal tract and abdomen were reviewed. Two histologically representative areas were identified and chosen for tissue microarray. Immunohistochemical staining was performed to demonstrate c-kit protein (CD117), CD34, smooth muscle actin, desmin and S-100 protein. The relations of various clinicopathologic features to outcome were analyzed.

RESULTSThe overall disease-specific survival of 194 patients was 93.5% at 1 year, 72.1% at 3 years and 63.2% at 5 years. Univariate analysis indicated that the tumor size, mitotic count, primary location, necrosis, high cellularity, mucosal invasion, mixed cell type, hemorrhage, direct tumor invasion of surrounding tissue, male sex, incompleteness of resection, cytologic atypia were significant predictors of survival. Multivariate analysis showed that tumor size, mitotic count, necrosis, direct tumor invasion of surrounding tissue and male sex were poor prognostic signs.

CONCLUSIONTumor size and mitotic count are important prognostic factors. However, to evaluate the prognosis of these tumors, a surgical pathologist should incorporate multiple parameters into their histologic evaluation in attempt to reach an appropriate opinion on the aggressiveness of GIST.

Aged ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; diagnosis ; mortality ; pathology ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Survival Rate