Objective To investigate the effects of apigenin on the migration of vascular smooth muscle cells (VSMC) induced by platelet derived growth factor (PDGF)-BB and the possible molecular mechanism. Methods VSMCs were isolated from thoracic aortas of male Sprague-Dawley rats using enzyme digestion method. Migration of VSMCs was determined by transwell assay. Western blotting was carried out to evaluate phosphorylation of c-jun N-terminal kinase (JNK). Results Treatment with PDGF-BB (20 ng·mL-1 ) significantly promote VSMC migration,the number of migrated cells was 2. 46 times than that of control group. However,after 12. 5 μmol·L-1 apigenin pretreatment,the number of migrated cells was 46. 5% of the PDGF-BB group. Various dose of apigenin can significantly inhibit VSMC migration induced by PDGF-BB,12. 5 μmol · L-1 apigenin treatment significantly inhibited PDGF-BB phosphorylation of JNK. Conclusion Apigenin can suppress the migration of VSMC induced by PDGF-BB. These beneficial effects on VSMC were at least partly mediated by the inhibition of activity of JNK.

Soil salinity is an important issue, as most crop plants are low in salt tolerance. Salt tolerance, a complex, multifactorial, and multigenic process, has been known to be a quantitative trait. The identification of the salt stress responsive genes or salt tolerance genes is essential for the breeding programs. Most recent efforts have been focused on the products of structural genes (transport proteins, ion channels, enzymes of solute synthesis) while little attention were paid to the regulatory aspects of these proteins. Since the first aquaporin gene from plants was cloned and functionally expressed in 1993, there has been a growing interest in the molecular biology of MIPs (membrane intrinsic proteins) and their bearing on the biophysics of water flow across plant membranes. In the last decades, studies on Mangroves, a special kind of wood plants, grow in high-salt and flooding conditions have been concentrated almost exclusively on their physiological and ecological characteristics. Kandelia candel, one of the dominant species of mangroves along the Chinese coast, lacks salt glands or salt hairs used for removal of excess salt in other mangroves. This makes K. candel a perfect model to study the molecular mechanism of salt tolerance in mangrove plants. Using cDNA RDA, a cDNA-specific modification of genomic representational difference analysis, a series of salt responsive genes of Kandelia candel were cloned. Among these gene fragments, a 183 bp fragment (termed as SRGKC1) encoding a tonoplast intrinsic protein (TIP) in Kandelia candel (KCTIP1) was identified. Based on the sequence of SRGKC1, two gene specific primers were designed, and the 3' and 5' end of the KCTIP1 gene were obtained using the SMART RACE cDNA Amplification Kit. RACE products were purified from low-melting agarose, and sequenced directly with GSPs as the sequencing primers. A 500-bp fragment corresponding to the 3'end of this gene was obtained using the GSP1 primer, and a 690 bp fragment corresponding to the 5' end of this gene was obtained using the GSP2 primer. Two primers that flank the putative open reading frame (ORF) were designed to obtain the cDNA containing the complete ORF by RACE PCR reaction. The full-length cDNA of KCTIP1, containing a 756 bp open reading frame (ORF), was approximately 1.1 kb; the start codon was located at the nucleotides of 99-101 and stop codon at the nucleotides of 855-857 followed by a poly (A) tail. The KCTIP1 cDNA sequence in this research was released in GenBank with accession number AF521135. Using ExPASy Proteomics tools provided by EMBL, the isoelectric point and MWt of KCTIP1 are estimated as 5.77 and 26.3 kD respectively. Transmembrane prediction analysis revealed the deduced KCTIP1 protein sequence contains six transmembrane regions at amino acid residues of 20 - 42, 57 - 79, 86 - 108, 113 - 135, 142 - 164 and 217 - 239. Two highly conserved asparagine-proline-alanine (NPA) motifs were located at 85 - 87 and 199 - 201 amino acid residues respectively. KCTIP1 is also predicted to contain the Cys residue (Cys 118) that are shown to confer Hg-sensitivity in Arabidopsis gamma-TIP and delta-TIP. Similarity analysis showed that KCTIP1 shared 77% - 79% amino acid sequence identity with the TIPs from Vitis berlandieri, Brassica oleracea and Arabidopsis thaliana. Expression analyses indicated that KCTIP1 had different expression among species of Mangroves. Expressions of KCTIP1 in Kandelia candel, Rhizophora apoculata and Ceriops tagal were suppressed by salt, and were insensitive to salt stress in unknown species of Mangroves. Previous studied showed that salt conditions might result in large and rapid changes in extracellular water potential and serious disturbance to the cytoplasm. In order to compensate for this imbalance, the relative contribution of water channels to flow across the root could thus vary. K. candel is a species that is native to intertial zone of tropical and subtropical coast and is well-adapted to salt conditions. The coordinated down-regulation of aquaporins in this plant may decrease membrane water permeability and thus increase the cellular water conserva- tion during periods of salt stress. The results reported here are consistent with the postulated roles for tonoplast water channels in regulating the hydraulic permeability of the vacuolar membranes and in adjusting the water homeostasis of the protoplasm under various physiological conditions. The identification of KCTIP1 as one of salt-responsive genes implies that intracellular osmotic equilibration is a part of salt-tolerant mechanisms in Mangroves.
Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; DNA, Complementary ; genetics ; Membrane Proteins ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Plant Proteins ; chemistry ; genetics ; Rhizophoraceae ; genetics ; Sequence Alignment

OBJECTIVETo investigate the prevalence of positive thyroid antibodies in children with type 1 diabetes mellitus (T1DM) and its influencing factors.

METHODSThe clinical data of T1DM children who were treated in the Children's Hospital of Zhejiang University from May 2005 to April 2011 were retrospectively studied. The relationships of thyroid globulin antibody (TGAb) and thyroid peroxidase antibody (TPOAb) with cytokines IL-2, IL-4, IL-6, IL-10, TNF and IFN-γ were evaluated, and the percentages of CD3+, CD4+ and CD8+ T-lymphocytes in peripheral blood were examined.

RESULTSA total of 186 T1DM children with complete data of both TGAb and TPOAb were included in the study, among whom 143 with normal TGAb and TPOAb levels and 43 (23.1%) presented with positive thyroid antibody (including 21 cases with both positive TGAb and positive TPOAb). Eighteen cases (9.7%) were diagnosed as autoimmune polyglandular syndrome type 3 variant (APS3v). Significantly more patients in the positive thyroid antibody group had a family history of diabetes than in the negative thyroid antibody group (27.9% vs 14.7%; P<0.05). The average age of the positive thyroid antibody group was 10.1±3.2 years, which was significantly greater than that in the negative thyroid antibody group (8.1±4.0 years) (P<0.05). The IL-2 level (4.48 ±1.27 pg/mL vs 2.82 ±0.84 pg/mL, P<0.05) and the percentage of peripheral CD3+ T-lymphocyte[(61±11)% vs (66±11)%; P<0.05] were also different between the positive and negative thyroid antibody groups.

CONCLUSIONSGenetic background and abnormal function of T-lymphocytes (especially higher IL-2 level) may be involved in the elevated prevalence of positive thyroid antibody in T1DM children.

Adolescent ; Autoantibodies ; blood ; Child ; Cytokines ; blood ; Diabetes Mellitus, Type 1 ; immunology ; Female ; Humans ; Male ; Polyendocrinopathies, Autoimmune ; etiology ; T-Lymphocytes ; immunology ; Thyroid Gland ; immunology

Objective:To explore the method and effect of free thining anterolateral thigh perforator flap (ALTP) in repairing soft tissue defect of scar contracture deformity after hand trauma.Methods:From March, 2015 to August, 2019, 12 patients who suffered scar contracture after hand trauma were repaired with thin ALTP. First, completely resected the scar contracture tissue from the hands and restored the normal bone structure and force line of the hand. The area of hand wound defects were 5.0 cm×6.0 cm-8.0 cm×10.0 cm. The wound was repaired by free ALTP, and the flap was micro-thinned for the first time. The flaps did not carry broad fascia, and the donor sites were directly sutured. The wound healing, the flap appearance, texture, sensation, scarring of the donor area, and functional recovery of the affected hand were observed regularly after surgery. The patients were followed-up by outpatient review and WeChat.Results:All the flaps survived well after the operation. Two cases suffered crisis because hematoma entraps vein cause by bleeding from perforator branch. After surgical exploration, the flaps survived successfully. All 12 flaps were followed-up successfully, including 6 cases reviewed in outpatient clinic, 4 cases followed by WeChat video and 2 cases by telephone consultation. The follow-up time was 3-20 months, with an average of 11 months. The flaps were not bloated, soft, non-pigmented, and beautiful in appearance. Only linear scars remained in the donor sites. The gripp function, palm function, thumb opposition function and finger function of the affected hand were largely restored. According to the TAM method of Upper Limb Function Evaluation of the Chinese Medical Association: 7 cases were excellent, 4 cases were good, and 1 case was fair.Conclusion:The thinning ALTP can be used to repair the scar contracture deformity after hand trauma. It can carry different tissues for 3-dimensional repair. After operation, the flap has a beautiful appearance, the donor site can be closed directly, and the damage of donor site can be reduced. It is an effective method to repair the hand contracture deformity.

OBJECTIVETo summarize lessons learned from an outbreak of severe acute respiratory syndrome (SARS) in China during the spring of 2004.

METHODSData of SARS cases were officially reported by Beijing Municipal Center for Disease Control and Prevention (BCDC) and Anhui Provincial Center for Disease Control and Prevention (APCDC) and results of epidemiological investigations were collected and analyzed.

RESULTSThree generations of 11 cases of SARS were identified during the outbreak. Initial two cases were most likely to be infected in Diarrhea Virus Laboratory of National Institute of Virology, China Centers for Disease Control and Prevention and main mode of transmission was direct contact with SARS patients. Delay in detecting initial case resulted in spread of the illness at hospitals and communities with two generations of secondary cases.

CONCLUSIONSSARS outbreak in 2004 has yielded following lessons for public health globally. (1) Lab bio-safety programs should be made and should be strictly abided by. Studies in highly pathogenic viruses such as SARS coronavirus should be utmost cautious. (2) Management systems of occupational exposure to virus and disease surveillance need to be strengthened to take all risk factors into account so as to detect potential patients with infectious disease as early as possible.

China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Occupational Exposure ; prevention & control ; Occupational Health ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; transmission

RecQ is a highly conserved helicase necessary for maintaining genome stability in all organisms. Genome comparison showed that a homologue of RecQ in Deinococcus radiodurans designated as DR1289 is a member of RecQ family with unusual domain arrangement: a helicase domain, an RecQ C-terminal domain, and surprisingly three HRDC domain repeats, whose function, however, remains obscure currently. Using an insertion deletion, we discovered that the DRRecQ mutation causes an increase in gamma radiation, hydroxyurea and mitomycine C and UV sensitivity. Using the shuttle plasmid pRADK, we complemented various domains of the D. radiodurans RecQ (DRRecQ) to the mutant in vivo. Results suggested that both the helicase and helicase-and-RNase-D-C-terminal (HRDC) domains are essential for complementing several phenotypes. The complementation and biochemical function of DRRecQ variants with different domains truncated in vitro suggested that both the helicase and three HRDC domains are necessary for RecQ functions in D. radiodurans, while three HRDC domains have a synergistic effect on the whole function. Our finding leads to the hypothesis that the RecF recombination pathway is likely a primary path of double strand break repair in this well-known radioresistant organism.
Amino Acid Sequence ; Deinococcus ; enzymology ; genetics ; Molecular Sequence Data ; Mutation ; genetics ; Phenotype ; Protein Structure, Tertiary ; RecQ Helicases ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid

OBJECTIVETo explore the effects of obesity on the peak level of luteinizing hormone (LH) in the gonadotropin-releasing hormone (GnRH) agonist test and obesity-related hormones in girls with central precocious puberty (CPP).

METHODSThree hundred and thirty-three girls with CPP who underwent the GnRH agonist test between 2012 and 2014 were classified into three groups: normal weight (n=123), overweight (n=108), and obesity (n=102), according to body mass index (BMI). The sexual development indices were compared between the three groups. Twenty girls were randomly selected from each group for evaluation of the serum levels of leptin, sex hormone binding globulin (SHBG), neurokinin B, and kisspeptin. The correlation of BMI with the levels of various hormones was assessed using Pearson correlation analysis.

RESULTSThere was no significant difference in mean age at diagnosis between the three groups; however, the bone age was significantly higher in the overweight and obesity groups than in the normal weight group (P<0.05). The peak level of LH in the GnRH agonist test and SHBG level in the normal weight group were significantly higher than those in the overweight and the obesity groups, while the serum levels of leptin and neurokinin B were significantly lower in the normal weight group than in the overweight and the obesity groups (P<0.05). BMI was negatively correlated with the peak level of LH in the GnRH agonist test and SHBG level (P<0.05), and positively correlated with the levels of leptin and neurokinin B (P<0.05).

CONCLUSIONSThe effects of BMI on the result of the GnRH agonist test and levels of obesity-related hormones should be taken into account in girls with precocious puberty.

Body Mass Index ; Child ; Female ; Gonadotropin-Releasing Hormone ; agonists ; Humans ; Leptin ; blood ; Luteinizing Hormone ; blood ; Neurokinin B ; blood ; Obesity ; blood ; Puberty, Precocious ; blood ; Sex Hormone-Binding Globulin ; analysis

OBJECTIVETo study the expression of transforming growth factor-β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) and its significance in premature infants with bronchopulmonary dysplasia (BPD).

METHODSA retrospective analysis was performed on the clinical data of 96 very low birth weight infants (gestational age of ≤ 32 weeks) who survived for more than 28 days and were admitted to the Neonatal Intensive Care Unit between January 2010 and December 2012. These subjects were divided into BPD group (n=21) and non-BPD group (n=75). The expression of TGF-β1 and PAI-1 in blood was measured by ELISA.

RESULTSThe levels of TGF-β1 and PAI-1 in the BPD group increased gradually from the 7th day to the 14th day and then to the 21st day after birth, and were significantly higher than in the non-BPD group at all time points (P<0.01). The TGF-β1 and PAI-1 levels in the non-BPD group on the 7th, 14th, and 21st days after birth were not significantly different from each other (P>0.05).

CONCLUSIONSThe expression of TGF-β1 and PAI-1 in blood is elevated in premature infants with BPD, which may be associated with the development of BPD.

Bronchopulmonary Dysplasia ; etiology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Plasminogen Activator Inhibitor 1 ; blood ; Retrospective Studies ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo investigate the drug resistance of pathogenic bacteria of nosocomial infections in the surgical intensive care unit.

METHODSThe drug resistance of pathogenic bacteria of nosocomial infections in the SICU in our hospital from January 2001 to December 2004 were analyzed.

RESULTSThe average nosocomial infections rate was 11.3%. The major sites of nosocomial infections were respiratory tract (30.9%), abdominal cavity (29.0%), bloodstream (9.7%) and biliary ducts (7.2%). The most common pathogens were pseudomonas aeruginosa (11.6%), methicillin-resistant coagulase negative staphylococci (11.1%) and candida albicans (9.7%). ESBLs-producing strains accounted for 66.2% and 58.5% of escherichia coli and klebsiella spp. respectively. Methicillin-resistant staphylococcus aureus accounted for 94.7% and methicillin-resistant coagulase negative staphylococci accounted for 88.2% in staphylococcus aureus and coagulase negative staphylococci. Carbapenems were the most powerful antibiotics against enterobacteriaceae. The non-fermenters were high resistant to antimicrobial agents. Vancomycin was the most potent antimicrobial against gram positive cocci. Amphotericin B was the most active antibiotic against fungi.

CONCLUSIONSMost strains of pathogens were antibiotic resistant in SICU. The main pathogenic bacteria of each infection site were different. So it is essential to establish nosocomial infections surveillance system in order to prevent, control and treat nosocomial infections effectively.

Bacterial Infections ; microbiology ; prevention & control ; Cross Infection ; microbiology ; prevention & control ; Drug Resistance, Bacterial ; Humans ; Intensive Care Units ; Microbial Sensitivity Tests

OBJECTIVETo develop a novel dual-modified vaccine, the superantigen-linked intestine-carcinoma cells expressing membrane-bound heat shock protein 70 (HSP70), and further examine its anticancer therapeutic effect.

METHODSThe pre-established intestine carcinoma CT26 line expressing membrane-bound heat shock protein 70 (HSP70) was amplified and incubated with superantigen fusion protein, staphylococcal enterotoxin A (SEA) fused with transmembrane sequence (SEA-TM), thereby the dual-modified vaccine was prepared after inactivation. The anticancer efficacy of the vaccine was examined.

RESULTSThe laser confocal microscopy and flow cytometry showed that there co-existed much HSP70 and SEA on the vaccine membrane surface. Both of the single-modified vaccines, the SEA-linked vaccine and membrane-bound-HSP70-expressing one, displayed marked tumor suppression, a prolonged survival period, augmented lymphocyte proliferation and higher NK and CTL activity in the vaccinated mice when compared with its counterpart. Furthermore, the dually modified vaccine induced lymphocyte proliferation most intensively, generated the highest NK and CTL activity as well as the strongest tumor rejection in the vaccinated mice. The survival period of the mice was further prolonged.

CONCLUSIONA new vaccine, SEA-linked and membrane-bound-HSP70-expressing intestine-carcinoma cells can induce more potent anticancer immunity and produce better therapeutic efficacy.

Animals ; Cancer Vaccines ; therapeutic use ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Enterotoxins ; immunology ; Gene Expression ; Genetic Vectors ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Superantigens ; immunology ; Transfection