Objective:To explore the mechanisms by which DNA methylation regulates miR-126 and its host gene EGFL7 in CD4+T cells from patients with systemic lupus erythematosus (SLE).
Methods:We analyzed the expression and the DNA methylation status within promoter region of EGFL7 and miR-126 by real-time qPCR and bisulifte genomic sequencing analysis.
Results:miR-126 and EGFL7 mRNA expression was upregulated in CD4+T cells from SLE compared with that from healthy controls (P<0.01). EGFL7 mRNA level was positively correlated with miR-126 expression in CD4+T cells from SLE (r=0.538, P=0.015). The average methylation level of EGFL7 promoter in CD4+T cells from SLE was lower than that from healthy controls (P<0.05).
Conclusion:hTe upregulation of miR-126 and its host gene EGFL7 expression in CD4+T cells from SLE is associated with the hypomethylation of the EGFL7 promoter.

Objective To investigate the relationship between the steroid-sensitive nephrotic syndrome(SSNS) and interleukin-18(IL-18) and to approach the inhibitive role of dexamethasone(DEX) on expression of IL-18 of peripheral blood mononuclear cells(PBMC) in children with SSNS in vitro.Methods IL-18 levels of serum, urine and supernatants of PBMC cultured in vitro were measured by enzyme linked immunosorbent assay(ELISA) in 23 children with SSNS who were either before or after treatment. Fifteen age-matched healthy children served as normal control group, and another 18 children with respiratory infections as infectious control group.Results There were signi-ficant differences of IL-18 in serum and urine before and after treatment in children with SSNS (t=15.072,16.149 Pa

AIMTo investigate the effects of RLI on plasma nitric oxide (NO) and NO synthase (NOS) isoforms of healthy humans.

METHODS30 healthy human subjects (aged from 40 - 70 years old) were recruited. RLI was induced by five 5 min cycles of ischemia of non dominant arm (200 mmHg, 5 min interval). Blood pressure, heart rate, and the feelings of ischemic arm were continuously monitored. Venous plasma was collected in contralateral arm at Pre, Post-0 h, Post-4 h, and Post-24 h. Plasma level of NO was measured by Griess reaction, and NOS was measured by chemical method.

RESULTSBlood pressure and heart rate varied in normal range. The uncomfortable feeling was decreased with the increasing numbers of ischemic cycles. Plasma level of NO, and iNOS in plasma were significantly increased at Post-0 h, Post-4 h, and Post-24 h compared to Pre (P < 0.05). tNOS was also significantly increased at Post-0 h and Post-4 h compared to Pre (P < 0.05). No significant change in plasma cNOS was shown at following three time points than Pre.

CONCLUSIONThese findings suggest that RLI can elevate plasma level of NO, tNOS, and iNOS in healthy humans. RLI might be a safe method as a rIPC, and it would have important possibility to be performed in clinic.

Adult ; Aged ; Arm ; blood supply ; Female ; Humans ; Ischemia ; blood ; physiopathology ; Ischemic Preconditioning ; methods ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; metabolism ; Reperfusion Injury ; physiopathology ; prevention & control

OBJECTIVENephrotic syndrome (NS) is characterized by marked urinary excretion of albumin and other intermediated-size plasma proteins such as transferrin. The aim of this study was to determine the changes of serum iron and transferrin and the relationship between the serum and urinary transferrin.

METHODSThe indexes related to iron metabolism, including serum iron, ferritin, transferrin, total iron-binding capacity, transferrin saturation and hematological parameters (Hb, MCV, MCH), and urinary transferrin were measured in 37 children with NS before treatment and at the remission stage. Thirty-five age-matched healthy children served as controls.

RESULTSSerum iron levels (18.8 +/- 3.8 micromol/L) in NS patients before treatment were significantly lower than in the healthy controls (22.2 +/-3.8 micromol/L) and those measured at the remission stage (21.0 +/- 3.5 micromol/L) (P < 0.01). Serum transferrin levels in NS patients before therapy (1.9 +/- 0.3 g/L) also decreased compared with those in the healthy controls (3.1 +/- 0.5 g/L) and those measured at the remission stage (2.9 +/- 0.6 g/L) (P < 0.01). In contrast, serum total iron-binding capacity and transferrin saturation were noticeably higher in NS patients before treatment than those in the healthy controls (total iron-binding capacity 56.4 +/- 9.2 micromol/L vs 50.7 +/- 6.8 micromol, P < 0.01; transferrin saturation 55.7 +/- 9.2 % vs 46.4 +/- 8.2%, P < 0.01) and were also higher than those measured at the remission stage (51.9 +/-7.7 micromol/L and 47.4 +/- 13.3%) (P < 0.01). Serum transferrin positively correlated to serum albumin (r = 0.609, P < 0.01) and negatively correlated to urinary transferrin (r = -0.550, P < 0.01) in NS patients before treatment.

CONCLUSIONSSerum iron and transferrin levels markedly decreased in NS patients, which may be partially related to the urinary loss of transferrin.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Iron ; blood ; Male ; Nephrotic Syndrome ; blood ; Transferrin ; analysis ; urine

Objective To summarize the workflow,strategy and experience of prenatal genetic test for deafness based on the 6-year clinical practice.Methods There were 213 famihes who received prenatal test from 2005 to 2011.Among the 213 families,205 families had had one deaf child,including 204 couples with normal hearing and one couple of the deaf husband and normal wife,8 families including 6 couples with normal hearing and 2 deaf couples,had no child before test. Genomic and mitochondrial DNA of each subject was extracted from whole blood.The etiology and recurrent risks in 212 families were confirmed by means of the genetic test of GJB2,SLC26A4 and mtDNA 12sRNA,but one family carried POU3F4 c.647G ＞ A heterozygous mutation causing X-linked hereditary hearing impairment confirmed by pedigree study.The prenatal test was carried out during the pregnancy of all mothers from 11 to 30 weeks,and the following genetic information and counseling were supplied based on the results.Results The recurrent risk was 25％in 209 families,including 204 families with one deaf child and 5 families without child,among which all couples were GJB2 or SLC26A4 mutation carriers and deaf children were caused by homozygous or compound GJB2/SLC26A4 mutations; The recurrent risk was 50％ in 3 families,the father and his child in one family had compound SLC26A4 mutations and the mother with heterozygous SLC26A4 mutation,the wife had POU3F4 c.647G ＞ A heterozygous mutation in another one family,and the husband with compound SLC26A4 mutations and the wife with mtDNA A1555G mutation and heterozygous SLC26A4 mutation simultaneously happened in the rest one family; The recurrent risk was 100％ in one family of the deaf couple who were both found to carry homozygous or compound GJB2 mutations,and the deaf wife got pregnant by artificial insemination with the sperm from the local Human Sperm Bank.226 times of prenatal test were applied in all 213 families that 11 families of them received prenatal test twice,and one family received three times. 46 times of prenatal testing showed that the fetuses carried parental mutations simultaneously or the same mutations with probands ; while 180 times of prenatal test showed that the fetuses carried only one parental mutation or did not carry any mutation from parents.The following visit showed that all of these 180 families had given birth to babies who were all revealed to have normal hearing by new born hearing screening test.Conclusions Prenatal diagnosis for deafness assisted by genetic test can provide efficient information about offspring's hearing condition,and the normative workflow and precise strategy highy guarantee the safe and favorable implementation of prental diagnosis.

AIMTo identify genes related to the human testis development by substrate hybridization technique.

METHODSA human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions. The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database. Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to determine the tissue expression profile of cul-3b.

RESULTSCul-3b, a novel CUL-3 transcript variant, was identified. The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones. Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites IRESes in the 5'-UTR. These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances. Additionally, cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR.

CONCLUSIONCul-3b is a novel transcript variant of CUL-3, which may be important not only for the development of human testis but also for that of other organs.

Base Sequence ; Cell Cycle Proteins ; genetics ; Cullin Proteins ; genetics ; Humans ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThis study aims to survey respiratory infectious disease related health literacy (RIDHL) and health behavior (RIDHB) among residents in Fengtai district, Beijing, analyze impact factors of RIDHL , explore the association between RIDHL and RIDHB.

METHODSMultistage sampling was employed and 1100 respondents were surveyed by self-designed questionnaires, which including social-demographic characteristics and evaluation of RIDHL and RIDHB. The survey results were described, the impact factors of RIDHL and the association between RIDHL and RIDHB were analyzed by analysis of variance or covariance.

RESULTSA total of 998 qualified questionnaires were recollected with the effective rate of 90.7%. The respondents aged from 15 to 65, scored (71.3 +/- 19.0) points in RIDHL test. Of those respondents, 25.7% (256/998), 43.2% (432/998) and 31.1% (310/998) were evaluated as low( <60 points), medium (60 - 85 points), and high level ( > 85 points) of RIDHL, respectively. There were significant difference in RIDHL scores between registered and non-registered residents, who scored (74.1 +/- 18.9) and (68.4 +/- 18.8) points, respectively (P < 0.01). RIDHL sections were ranked as audiovisual (77.6%, 4647/5988), internet using (75.2%, 2251/2994), reading (74.6%, 3724/4990), map using (68.3%, 4090/5988) and quantitative (65.5%, 5230/7984) according to the accurate rates from high to low. Analysis of variance or covariance showed that RIDHL scores were significantly different among respondents with different ages, nationalities, educational levels, occupations, and incomes (P < 0.01), yet no significant differences among those with different genders and marital status (P > 0.05). Respondents scored (69.7 +/- 15.5) points in RIDHB test. The RIDHB scores ((64.5 +/- 15.0), (70.4 +/- 15.6), (72.5 +/- 14.9) points, respectively) increased among residents with low, medium and high level of RIDHL (P < 0.01).

CONCLUSIONResidents in Fengtai district, Beijing possessed medium level of RIDHL. The non-registered residents showed lower RIDHL than registered residents. Ages, nationalities, educational levels, occupations, and incomes were impact factors of RIDHL. People with higher level of RIDHL also showed a higher level of RIDHB.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Communicable Diseases ; epidemiology ; Female ; Health Behavior ; Health Literacy ; Humans ; Male ; Middle Aged ; Respiratory Tract Infections ; epidemiology ; prevention & control ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo explore the role of reversal multidrug resistance (MDR) using short hairpin RNA (shRNA) expression vectors in multidrug resistance human leukemia cell line K562/ADM.

METHODSThe oligonucleotides with 19-mer hairpin structure were synthesized. The shRNA expression vectors were constructed and introduced into K562/ADM cells. Expression of mdr1 mRNA was assessed by RT-PCR, and P-gp expression was determined by Western blot. The apoptosis and sensitivity of the K562/ADM cells to doxorubicin were quantified by flow cytometry and methyl thiazolyl tetrazolium (MTT) assays, respectively. Cellular daunorubicin accumulation was assayed by laser confocal scanning microscope (LCSM).

RESULTSIn positive clones of K562/ADM cells stably transfected with pSilencer 3.1-HI neo mdr1-A and mdr1-B shRNA expression vectors, RT-PCR showed that mdr1 mRNA expression was significantly reduced to 35.9% (P < 0.05), 27.5% (P < 0.01), respectively. Western blot showed that P-gp expression was significantly and specifically inhibited. Resistance against doxorubicin was decreased from 79-fold to 38-fold (P < 0.05), 30-fold (P < 0.01) respectively. Furthermore, the fluorescence intensity of K562/ADM cells was increased significantly compared with the control. shRNA vectors significantly enhanced the cellular daunorubicin accumulation. The percent of the apoptosis cell was significantly enhanced to 18.1% (P < 0.05) , 54.4% (P < 0.01) respectively.

CONCLUSIONSshRNA expression vectors can effectively reverse MDR, and restore the sensitivity of drug-resistance K562/ADM cells to conventional chemotherapeutic agents.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

OBJECTIVEThis study sought to isolate and identify the proteins that interact with ataxin-3, to confirm the interacted domain, and to provide new clues for exploring the function of ataxin-3 and the pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD).

METHODSYeast two-hybrid screen (MATCHMAKER GAL4 Two-Hybrid System 3) and regular molecular biologic techniques were undertaken to screen human brain cDNA library with mutant ataxin-3 bait. Two baits from both normal and mutant C-terminus of ataxin-3 were created by subcloned methods to determine which domain of ataxin-3 interacts with the putative associated proteins and to find out optimal candidate proteins that interact with C-terminus of ataxin-3. Confocal microscope was used to observe whether ataxin-3 co-localized with the obtained interacting proteins in mammalian cells.

RESULTSFive novel ataxin-3 interacting proteins were obtained, among which were three known proteins, namely human rhodopsin guanosine diphosphate dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2; the other two were unknown. Interacting domain analysis revealed that an unknown protein interacted with the C-terminus near the polyglutamine tract of ataxin-3, the other four all interacted with the N-terminus. In the nucleus of SH-SY5Y cell, small ubiquitin-like modifier 1 co-localized with the wild-type ataxin-3 and with the intranuclear aggregates formed by the mutant ataxin-3.

CONCLUSIONAn unknown protein probably interacting with C-terminus of ataxin-3 is firstly discovered, and the initiative findings suggest first that the interaction of small ubiquitin-like modifier 1 with N-terminus of ataxin-3 and the relevant sumoylation probably participate in the post-translation modifying of ataxin-3 and in the pathogenesis of SCA3/MJD.

Acid Sensing Ion Channels ; Ataxin-3 ; Cell Line, Tumor ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Confocal ; Mutation ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; SUMO-1 Protein ; genetics ; metabolism ; Sodium Channels ; genetics ; metabolism ; Transfection ; Two-Hybrid System Techniques

Adolescent ; Adult ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use ; Severity of Illness Index ; Treatment Outcome