OBJECTIVETo observe the hair follicle regeneration after subcutaneous implantation of hair follicle cell mixture in nude mice.

METHODSThe hair papilla cells, dermal sheath cells, outer root sheath and fibroblasts of human scalp were mixed with the hair follicle epithelial cells and implanted subcutaneously in nude mice to observe the regeneration of the hair follicle.

RESULTS AND CONCLUSIONFormation of intact hair follicle-like structures was observed in the skin sections of the recipient nude mice, suggesting the feasibility of this approach for hair follicle regeneration in vivo.

Animals ; Cell Transplantation ; Hair Follicle ; cytology ; metabolism ; physiology ; transplantation ; Humans ; Injections, Subcutaneous ; Mice ; Mice, Nude ; Regeneration ; Skin Pigmentation ; Time Factors

Background:Andersson lesions (ALs),also known as spondylodiscities,destructive vertebral lesions and spinal pseudarthrosis,usually occur in patients with ankylosing spondylitis (AS).Inflammatory and traumatic causes have been proposed to define this lesion.Different surgical approaches including anterior,posterior,and combined anterior and posterior procedure have been used to address the complications,consisting of mechanical pain,kyphotic deformity,and neurologic deficits.However,the preferred surgical procedure remains controversial.The aim of this study was to illustrate the safety,efficacy,and feasibility of a modified posterior wedge osteotomy for the ALs with kyphotic deformity in AS.Methods:From June 2008 to January 2013,23 patients (18 males,5 females) at an average age of 44.8 years (range 25-69 years) were surgically treated for thoracolumbar kyphosis with ALs in AS via a modified posterior wedge osteotomy in our department.All sagittal balance parameters were assessed by standing lateral radiography of the whole spine before surgery and during the follow-up period.Assessment of radiologic fusion at follow-up was based on the Bridwell interbody fusion grading system.Ankylosing spondylitis quality of life (ASQoL) and visual analog scale (VAS) scores were performed to evaluate improvements in daily life function and back pain pre-operatively and post-operatively.Paired t tests were used to compare clinical data change in parametric values before and after surgery and the Mann-Whitney U test was employed for non-parametric comparisons.The radiographic data change was evaluated by repeated measure analysis of variance.Results:The mean operative duration was 205.4 min (range 115-375 min),with an average blood loss of 488.5 mL (range 215-880 mL).Radiographical and clinical outcomes were assessed after a mean of 61.4 months of follow-up.The VAS back pain and ASQoL scores improved significantly in all patients (7.52 ± 1.31 vs.1.70 ± 0.70,t=18.30,P ＜ 0.001;13.87 ± 1.89 vs.7.22 ± 1.24,t=18.53,P＜0.001,respectively).The thoracolumbar kyphosis (TLK) changed from 40.03±17.61° pre-operatively to 13.86 ± 6.65° post-operatively,and 28.45 ± 6.63° at final follow-up (F =57.54,P ＜ 0.001),the thoracic kyphosis (TK) changed from 52.30 ± 17.62° pre-operatively to 27.76 ± 6.50° post-operatively,and 28.45 ± 6.63° at final follow-up (F =57.29,P ＜ 0.001),and lumbar lordosis (LL) changed from-29.56 ± 9.73° pre-operatively to-20.58 ± 9.71° post-operatively,and-20.73 ± 10.27° at final follow-up (F=42.50,P ＜ 0.001).Mean sagirtal vertical axis (SVA) was improved from 11.82 ± 4.55 cm pre-operatively to 5.12 ± 2.42 cm post-operatively,and 5.03 ± 2.29 cm at final follow-up (F=79.36,P ＜ 0.001).No obvious loss of correction occurred,according to the lack of significant differences in the sagittal balance parameters between post-operatively and the final follow-up in all patients (TK:27.76 ± 6.50° vs.28.45 ± 6.63°,TLK:13.86 ± 6.65° vs.14.42 ± 6.7°,LL:-20.58 ± 9.71° vs.-20.73 ± 10.27°,and SVA:5.12 ± 2.42 cm vs.5.03 ± 2.29 cm,all P ＞ 0.05,respectively).Conclusions:The modified posterior wedge osteotomy is an accepted surgical procedure for treating thoracolumbar kyphosis with ALs in AS and results in satisfactory local kyphosis correction,solid fusion,and good clinical outcomes.

OBJECTIVETo acquire sufficient PDGF-BB protein and provide the basis for the further studies of its role on the fracture healing and trauma reconstruction and its clinical applications.

METHODSConstructed the prokaryotic expression vector pQE-PDGF-B with the gene rearrangement technique, and the monomeric form of recombinant PDGF-B expressed in E. coli M15.

RESULTSPDGF-B mature peptide gene was inserted into prokaryotic expression vector pQE30, which was confirmed by PCR, enzyme digestion and sequencing identification; the expressed products of pQE-PDGF-B in E. coli showed a single protein on SDS-PAGE, and their expression level was about 15% of the total bacterial protein. The molecular weight of the purified PDGF-B protein was about 15 KDs on SDS-PAGE.

CONCLUSIONSThe construction of recombinant plasmid and preparation of the monomeric protein of PDGF-B provides a solid foundation for further studying the function of PDGF-BB and producing biologically PDGF-BB protein.

Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; In Vitro Techniques ; Platelet-Derived Growth Factor ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-sis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

This article reported the clinical manifestations, steroid profiles and adrenal ultrasound findings in two unrelated Chinese girls with lipoid congenital adrenal hyperplasia (LCAH). Direct DNA sequencing and restriction fragment length polymorphism (RFLP) analysis were used to identify the mutations of steroidogenic acute regulatory protein (StAR) gene. The two patients with 46,XX karyotype, presented hyperpigmentation, growth retardation, and hyponatremia. Steroid profiles analysis revealed elevated plasma adrenocorticotrophic hormone levels, decreased or normal serum cortisol levels and low levels of androgens. Ultrasound examinations revealed that enlarged adrenals in patient 1 and normal adrenals in patient 2. Direct DNA sequencing of StAR gene showed a reported homozygous for c.772C>T(p.Q258X) in patient 1. Compound heterozygous for c.367G>A(p.E123K) and IVS4+2T>A (both novel mutations) were found in patient 2, inherited from her mother and father respectively. The amino acid of mutant position of the novel p.E123K was highly conserved in ten different species and was predicted to have impacts on the structure and function of StAR protein by the PolyPhen-2 prediction software. RFLP analysis revealed three bands (670, 423 and 247 bp) in patient 2 and her father and two bands (423 and 247 bp) in her mother and 50 controls. It is concluded that LCAH should be considered in girls with early onset of adrenal insufficiency and that steroid profiles, karyotype analysis, adrenal ultrasound and StAR gene analysis may be helpful for the definite diagnosis of LCAH.
46, XY Disorders of Sex Development ; diagnosis ; genetics ; Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Amino Acid Sequence ; Child ; Female ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Mutation ; Phosphoproteins ; genetics ; Polymorphism, Restriction Fragment Length

In order to differently diagnose avian influenza virus (AIV) subtypes, the HA gene of AIV H9 subtype was cloned, expressed and utilized in an enzyme-linked immunoad sorbent assay (ELISA). HA gene (1683bp) of H9N2 AIV was amplified by RT-PCR from a strain of field isolated H9N2 AIV, and its identity was confirmed by sequencing. The HA gene was subcloned into prokaryotic expression vector pGEX-KG with its secretion signal sequence removed. The expressed HA-GST fusion protein in E. coli BL21 was characterized by SDS-PAGE and western blotting analysis as a 90kD protein with immunogenicity. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation. The HA-GST fusion protein was used to establish an indirect ELISA for the detection of antibodies to H9 subtypes of AIV. The assay has 91.57% specificity to H9 AIV, 92.31% sensitivity and excellent reduplication. It could be used to differently detect antibodies to H9 AIV.
Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; Humans ; Influenza A Virus, H9N2 Subtype ; genetics ; Influenza, Human ; diagnosis ; virology ; Recombinant Proteins ; biosynthesis ; genetics

AIM:To investigate the effects of kaempferol-3-O-rutinoside(KR)on the proliferation,migration of vascular smooth muscle cells(VSMC)and the activation of transforming growth factor βreceptor 1(TGFBR1)signaling pathway in the cells.METHODS: The viability of VSMC was detected by MTT assay.The proliferation of VSMC was measured by EdU staining.The migration ability of VSMC was examined by Transwell assay.The protein levels of the mi-gration-associated proteins matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)were detected by Western blot.Molecular docking study was conducted to explore the interaction between KR and TGFBR 1.The protein le-vels of the phosphorylated TGFBR1,Smad2 and Smad3 were determined by Western blot.RESULTS: KR inhibited the viability of VSMC in a dose-and time-dependent manner.KR reduced the ratio of EdU-positive cells in a dose-dependent manner.KR dose-dependently suppressed the migration ability of VSMC and decreased the protein levels of MMP 2 and MMP9(P<0.05).KR docked into TGFBR1 with the binding energy of -9.804 kcal/mol by forming hydrogen bonds with SER-280,ARG-215,ASP-290 and LYS-335 of TGBFR1.KR dose-dependently suppressed the activation of TGFBR 1 and its downstream proteins Smad2 and Smad3(P<0.05).CONCLUSION: KR inhibits the proliferation and migration of VSMC,possibly via blocking the TGFBR1 signaling pathway.