Inflammation-mediated age-dependent effects of casein kinase 2-interacting protein-1 on osteogenesis in mesenchymal stem cells / 中华医学杂志(英文版)

BACKGROUND@#The casein kinase 2-interacting protein-1 (CKIP-1) is important in the development of osteoblasts and cardiomyocytes. However, the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells (MSCs) remain unclear. This study aimed to determine whether CKIP-1 affects osteogenic differentiation in MSCs and explore the relationship of CKIP-1 and inflammation.@*METHODS@#Bone marrow MSCs of CKIP-1 wild type (WT) and knockout (KO) mice were cultivated in vitro. Cell phenotype was analyzed by flow cytometry, colony formation was detected to study the proliferative ability. Osteogenic and adipogenic induction were performed. The osteogenic ability was explored by alizarin red staining, alkaline phosphatase (ALP) staining and ALP activity detection. Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the mRNA expression levels of osteoblast marker genes. The adipogenic ability was detected by oil red O staining. Content of the bone was analyzed to observe the differences of bone imaging parameters including trabecular bone volume/tissue volume (BV/TV), bone surface area fraction/trabecular BV, trabecular number (Tb.N), and trabecular spacing (Tb.sp). Interleukin (IL)-1β was injected on WT mice of 2 months old and 18 months old, respectively. Difference in CKIP-1 expression was detected by RT-PCR and western blot. The relationship between CKIP-1 and inflammation was explored by RT-PCR and western blot.@*RESULTS@#ALP assays, alizarin red staining, and qRT-PCR showed that MSCs derived from CKIP-1 KO mice exhibited a stronger capability for osteogenesis. Micro-computed tomography detection showed that among 18-month-old mice, CKIP-1 KO mice presented significantly higher bone mass compared with WT mice (P = 0.02). No significant difference was observed in 2-month-old mice. In vivo data showed that expression of CKIP-1 was higher in the bone marrow of aging mice than in young mice (4.3-fold increase at the mRNA level, P = 0.04). Finally, the expression levels of CKIP-1 in bone marrow (3.2-fold increase at the mRNA level, P = 0.03) and cultured MSCs were up-regulated on chronic inflammatory stimulation by IL-1β.@*CONCLUSIONS@#CKIP-1 is responsible for negative regulation of MSC osteogenesis with age-dependent effects. Increasing levels of inflammation with aging may be the primary factor responsible for higher expression levels of CKIP-1 but may not necessarily affect MSC aging.

A modified preauricular-temporal approach for fixing comminuted and redisplaced zygomatic arch fractures with the resorbable bone plate / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the effectiveness of the rigid internal fixation for comminuted and redisplaced zygomatic arch fractures by modified preauricular-temporal approach with the resorbable bone fixation.</p><p><b>METHODS</b>Totally twenty patients aged from 14 to 68 years and admitted to our hospital between September 2006 and June 2011 were reviewed, of whom seventeen had a unilateral comminuted zygomatic arch fracture and three re-displaced arch fracture after failed closed reduction. The fracture segments were aligned to restore the preinjury form of the arch by rigid fixation with resorbable plates and screws through a modified preauricular-temporal incision.</p><p><b>RESULTS</b>The fractures were well reduced, preauricular-temporal scar and lateral facial contour were aesthetically satisfying, and no case had limited mouth opening as well as facial palsy. The resorbable plates were not palpated one year after the operation.</p><p><b>CONCLUSION</b>The rigid internal fixation through the preauricular-temporal approach with the resorbable bone is an effective method for the comminuted and redisplaced zygomatic arch fractures.</p>

The preliminary study of the use of MRI navigation in identifying the safe surgical margin of the maxillofacial malignancy / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility of MRI navigation in identifing the safe surgical margin of the maxillofacial malignancy.</p><p><b>METHODS</b>The pathology results of the surgical margin identified by the technique of MRI navigation form 20 patients with maxillofacial malignancy were compared with those of 45 patients with maxillofacial malignancy who underwent the routine operation without MRI navigation.</p><p><b>RESULTS</b>There was no difference between the two groups of patients in age, sex, size of tumor, tumor stages, pathologic diagnosis (P > 0.05). The negative rate of the surgical margin of the lesions treated by surgery with the technique of MRI navigation was significantly lower than that of the lesions treated without MRI navigation (P = 0.007) and highly correspondent with the pathology results.</p><p><b>CONCLUSIONS</b>MRI navigation was helpful in identifying the safe surgical margin of the maxillofacial malignancy.</p>

Experiment on inducing human periodontal ligament stem cells into adipose cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the capability of human periodontal ligament stem cells (PDLSCs) differentiating into adipose cells in vitro and to determine their changes in cell morphology, structure and function during differentiation.</p><p><b>METHODS</b>PDLSCs isolated by magnetic-activated cell selection were treated continuously with adipogenic medium for 21 d. Then the cell morphology, ultrastructure, adipose specific markers of low density lipoprotein (LPL) and peroxisome proliferator activated receptor-gamma (PPAR-gamma) were analyzed by inverted contrast microscope, trans mission electron microscope (TEM), flow cytometry, immunofluorescence, RT-PCR and Western blot, respectively. These adipose-like cells were also identified by oil red O staining to determine the formation of lipid droplet, and the non-induced cells were used as control.</p><p><b>RESULTS</b>After continuous induction, the treated cells differentiated into adipose-like cells with round shape, and large amount of lipid drop in cytoplasm. 96.54% of the PDLSCs were found to differentiate into adipose cells as showed by flow cytometry, the specific markers of LPL mRNA and PPAR-gamma mRNA, and oil red O staining, respectively. Further, PPAR-gamma protein was detected in the induced cells in a time-dependent manner.</p><p><b>CONCLUSION</b>Human PDLSCs have the potential of differentiating into adipose cells under appropriate condition, and the differentiated cells exhibited characteristics of adipose cells both from cell morphology and from their functions.</p>

Three-dimensional finite element analysis for different directions distraction at midface / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the biomechanical changes with different directions distraction at midface.</p><p><b>METHODS</b>An anteriorly directed 500 g force was applied to the floor of apertura piriforms in different directions to the occlusal plane. Three-dimensional finite element analysis was used to evaluate the biomechanical change of craniofacial complex.</p><p><b>RESULTS</b>As the force direction was moved downward, the sagittal distraction length of the craniofacial complex decreased and vertical movement changed from upward to downward. The craniofacial complex was moved anteriorly when the downward force was applied about 20-30 degrees to the occlusal plane. The forces could generate the uniform stress distribution in the craniofacial sutures and avoid counterclockwise rotation of the maxilla.</p><p><b>CONCLUSIONS</b>The craniofacial complex can be effectively distracted anteriorly when the downward force is applied to the floor of aperture piriforms in direction of 20-30 degrees to the occlusal plane.</p>

Preparation of the immunotoxin 2E8-norcantharidin and its targeting killing effect in vitro / 中华儿科杂志

<p><b>OBJECTIVE</b>Monoclonal antibody (mAb) conjugated with certain toxin to generate immunotoxin bears an important and promising effect as a new therapy for patients with hematopoietic malignancies. However, most toxic moieties conjugated with antibody proteins reported in the literature were toxic proteins which presented immunogenicity to patients capable of inducing anti-toxin antibody. Norcantharidin (NCTD) is a small molecule toxin. It does not have the immunogenicity to human body so that it bears a promising potential for development of new targeting drug. In this study, a new clone of self-made anti-CD19 mAb named ZCH-4-2E8 conjugated with NCTD was used to investigate its targeting efficacy against CD19+ lymphoid malignant Nalm-6 cells in vitro to provide the experimental data for the further development of this new targeting agent.</p><p><b>METHODS</b>A monoclonal antibody named 2E8 was prepared from mouse ascites and purified by gel chromatography. The purity of the antibody protein was checked by SDS-PAGE assay. Immunotoxin 2E8-NCTD was successfully generated through conjugating CD19 mAb protein and Norcantharidin by the activated ester method. The binding activity of the immunoconjugate (2E8-NCTD) against CD19 antigens on cell surface and the expression levels of CD19 antigens on Nalm-6 and K562 cells were examined by flow cytometry. Comparisons of the inhibitory effects among PBS, purified 2E8 antibody, norcantharidin and immunotoxin 2E8-NCTD groups on cell growth of either Nalm-6 cells or K562 cells were made.</p><p><b>RESULTS</b>The purity of the purified 2E8 antibody was higher than 99.00% demonstrated by SDS-PAGE assay. 2E8 antibody in the supernatant reacted with 99.34% of Nalm-6 cells, while only 0.98% of K562 cells reacted with this antibody. The newly generated immunotoxin (2E8-NCTD) had a positive rate of 99.90% on Nalm-6 cells with little reduction of binding activity. From the in vitro study, both 2E8-NCTD and norcantharidin were shown to have significant inhibitory effects on the growth of CD19+ Nalm-6 cells (P < 0.001), while the purified 2E8 antibody did not show any significant influences on the growth of Nalm-6 cells. Since no significant inhibitory effects were identified among immunotoxin 2E8-NCTD, 2E8 antibody and control groups on CD19(-) K562 cells, a significant targeting effect of the 2E8-NCTD against Nalm-6 cells was confirmed.</p><p><b>CONCLUSIONS</b>The immunotoxin 2E8-NCTD was successfully synthesized by activated ester method with an excellent targeting killing effect on CD19+ Nalm-6 leukemia cells in vitro, which provides some experimental data for the further development of this new targeting agent.</p>

Three-dimensional finite element analysis of craniofacial skeleton protraction at medium position / 中华整形外科杂志

<p><b>OBJECTIVE</b>The purpose of this study was to investigate the biomechanical changes of midface skeleton protraction at its medium position in the craniofacial complex, using the three-dimensional finite element method (FEM).</p><p><b>METHODS</b>A three-dimensional FEM model was developed from the CT scan images by the technologies of three-dimensional reconstruction, image processing and meshing. The protraction forces were applied to the following locations: the first molar, full maxillary arch, and the floor of aperture piriforms. Biomechanical changes from different position protraction were investigated by means of finite element analyses.</p><p><b>RESULTS</b>Protraction forces at the level of the floor of aperture piriforms produced a more forward movement of the upper maxilla in sagittal direction. Vertical and lateral displacements were less than those in loading with teeth or denture. Compressive stress on the radix nasi decreased obviously in midface skeleton protraction at its medium position.</p><p><b>CONCLUSIONS</b>Compared with traditional orthopedic protraction, midface skeleton protraction at its medium position could advance maxilla en bloc, decrease the counterclockwise rotation of the maxilla, and reduce the constriction of the anterior part of the palate.</p>

Effects of testosterone and cortisol in sports fatigued organism by Chinese material medica of shixiang yaofa / 中国中药杂志

<p><b>OBJECTIVE</b>To study the endogenous hormone of testosterone and cortisol that secretes volume and rhythm in sports fatigued human bodies and animals. To determine secretory volume and rhythm in sports fatigued human bodies and animals when Shixiang yaofa's drug are used.</p><p><b>METHOD</b>Radio-immunity method was used to determine secretory volume and rhythm in sports fatigued human bodies and animals and to analyze secretory volume and rhythm. According to the secretory volume and rhythm of testosterone and cortisol, the Shixiang Yaofa's drugs were used. Doses: wenyang jihuo bead 10 g/sack 2 sack, ziyin xiuyang capsule 0.5 g/pill 8 pill pd in human bodies for 28 days. Wenyang jihuo bead 10 g x kg(-1) of crude drug, ziyin xiuyang capsule 4 g x kg(-1) of crude drug pd for hight doses in animals. Other groups for low doses 5 g x kg(-1) and 2 g x kg(-1) of crude drug pd for 15 days. The blood samples were collected for determination.</p><p><b>RESULT</b>(1) In human bodies the peak value of testosterone was appeared in 8:00 and valley value was appeared in 18:00, ranges: 176.93-281.73 x 10(-5) mg x L(-1). The peak value of cortisol was appeared in 8:00 and valley value was appeared in 22:00, ranges: 1.31-16.13 x 10(-3) mg x L(-1). In the same condition, the mouse peak value of testosterone was appeared in 20:00 and valley value was appeared in 0:00, ranges: (0.56 x 10(-5) - 124.0 x 10(-5)) mg x L(-1). The rhythm in animals was compatible with human bodies, howerer, the peak value was delayed for 12 hours. (2) The testosterone was step up and the cortisol was cut down in sports fatigued human bodies and animals when shixiang yaofa's drug were used (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>The secretion of testosterone and cortisol in sports fatigued human bodies and animals are existed conclusive biologic rhythm. The secretory volume can be available accommodation by shixiang yaofa's drugs.</p>

Establishment of the database of female skull by the aesthetics / 中华整形外科杂志

<p><b>OBJECTIVE</b>The three-dimensional (3D) craniofacial measurements were studied through the quantitative computed tomography (CT). The dynamic database of quantitative measurement of three-dimensional craniofacial bone was established as mandible in physiological position.</p><p><b>METHODS</b>170 aesthetics female were examined by spiral volumetric CT (GE SR-7000). 3D craniofacial bone images were reformatted and 3D measurements were performed in SUN Workstation respectively. 33 points were defined in the 3-d craniofacial structure in screen, 14 distances and 11 angles were measured, and 12 ratios were calculated in each case. All data were transferred into the database based on the SPSS software. There is all information of one case (such as number, sex, age, distances, angers) in one row; each column is a measurement item. The mean, standard deviation, standard error, medium, coefficient of variation and 95% confidence interval of data can be calculated and the correlation, regression between several groups of measurement item can be proceeded by computer automatically in the dynamic database.</p><p><b>RESULTS</b>3D craniofacial bone imagings were displayed in arbitrary views without disturbing superposition by using cutting, rotating and 3D measurement procedures. The large data volume provides more information of special relationship of skull base, zygomatic bone, maxilla, mandible and vertebra. The coefficient of variation of skull base is less than them of maxilla and mandible. The standard deviation of ratios is further smaller than the standard deviation of distances and angles. With stepwise regression, the equation is (Go - Go) Y = 0.578X1 + 0.754X2 + 0.228X3 - 0.579X4 - 14.672; (Tz- Tz) : Y = 0.775X1 + 0.161X2 + 0.348X3 + 0.201X4 + 27.730.</p><p><b>CONCLUSIONS</b>The database offers reference of the studying of growth rule of craniofacial bone of aesthetics female. It will help improve diagnostic accuracy, staging of reconstruction, precision of corrective surgery, and follow-up patients.</p>

Reactivity of a novel monoclonal antibody ZCH-2B8a on normal hematopoietic cells and malignant cell lines and its significance / 中国实验血液学杂志

ZCH-2B8a (IgG2a) is a novel monoclonal antibody (McAb) generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KG1a as immunogen. This antibody has been submitted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA8) and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells, natural killers (NK), neutrophils, monocytes, dendritic cells (DC), red blood cells (RBC), platelets (Plt), hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34(+) cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells > or = 20% was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29% and 2/3 specimens of monocytes with an average positive rate of 59.84%. 2B8a was weakly reactive to neutrophils (23.72%) and negative for T cells, NK, DC, RBC and Plt. The antibody reacted to all 3 marrow CD34(+) cells with an average positive rate of 39.33% while it was negative for G-CSF-mobilized CD34(+) peripheral blood stem/progenitor cells (PBSC, 1.25%). Cell line analysis showed that the antibody notably reacted to three out of 4 cell lines (Raji, SMS-SB, Nalm-6 and Nall-1) with the positive rates of 98.78%, 98.61%, 94.93% respectively and weakly to one of them with 5.68% in B lineage cell lines and monoblastic cell line (U937, 67.78%) while it was only weakly positive or negative for other myeloid leukemia cell lines including Meg01 (33.40%), HL-60 (29.70%), K562 (28.19%), KG1a (16.23%) and HEL92.1.7 (8.02%). Among 4 T lineage leukemia, 5 neuroblastoma and 1 colon cancer cell lines tested, only Molt-3 was found weakly positive (31.40%) for 2B8a, while the remaining 3 T cell lines (Molt4, JM and CCRF-CEM), 5 neuroblastoma cell lines (LA-N1, KCNR, BE, SK-N-SH, SK-N-AS) and the colon cancer cell line (HR8348) tested were negative. An amniotic epithelium cell line (FL) was showed positive for the antibody (45.03%). It is concluded that 2B8a antibody primarily reacts to B lineage and monocytic lineage cells which may bear the diagnostic and therapeutic applications among different types of hematopoietic malignancies.

Reactivity of a novel monoclonal antibody ZCH-7-2D3 on leukemia cells and its clinical significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To analyze the reactive pattern and its clinical significance of ZCH-7-2D3 monoclonal antibody on leukemia cells.</p><p><b>METHODS</b>Bone marrow samples from 100 leukemia patients (male 59: female 41, 34 cases of children and 66 adults, aged 11 months - 77 year) were collected. A CD45 gating strategy and multi-parameter flow cytometry were used to analyze the leukemia cells.</p><p><b>RESULT</b>The positive rate (10/31) of 2D3 antibody on acute lymphocytic leukemia (ALL) patients was significantly lower than that (39/55) of acute myelogeneous leukemia (AML) (P <0.01). 2D3 was positive for all three cases of B/myeloid mixed lineage leukemia, but negative in 6 cases of chronic myelogeneous leukemia (CML), significantly lower than that in AML (39/55, P<0.01) patients. There was no difference between the positive rates of chronic lymphocytic leukemia (CLL, 3/5) and B lineage ALL (9/28, P = 0.2389). The antibody was not found to recognize the differentiation stages of either ALL and AML subtypes.</p><p><b>CONCLUSION</b>2D3 monoclonal antibody primarily recognizes AML cases. Further studies on cloning of gene encoding the antigen protein and its biological functions are warranted.</p>

Suppositional sutural construction and application in the three-dimensional finite element model of craniofacial complex / 中华整形外科杂志

<p><b>OBJECTIVE</b>The purpose of this study was to explore suppositional sutural construction and protraction in the three-dimensional finite element model of craniofacial complex.</p><p><b>METHODS</b>Combining spiral CT scanning technology with the three-dimensional finite element method, three-dimensional FEM model of craniofacial complex was developed for analysis. In the model, craniofacial sutures were constructed through the MSC. Patran program and different mechanical properties of sutures were applied. Under the same loading condition, bone displacements of the maxilla were calculated.</p><p><b>RESULTS</b>A three-dimensional FE model of craniofacial complex, comprising teeth and craniofacial sutures, was developed. The displacement of the skeletal structures with sutures was significantly larger than that without sutures.</p><p><b>CONCLUSIONS</b>The construction of craniofacial sutures in the three-dimensional finite element model of craniofacial complex could improve the accuracy of finite element model, which set up good foundation for biomechanical studies of craniofacial complex.</p>

Trans-sutural distraction osteogenesis for early correction of midfacial hypoplasia in children: a primary clinical report / 中华整形外科杂志

<p><b>OBJECTIVE</b>To probe the possibility of trans-sutural distraction osteogenesis for correction of children midfacial hypoplasia.</p><p><b>METHODS</b>The trans-sutural distraction system of mid-facial skeleton consisted of the bone-borne traction hooks of titanium, the face-bow, and the elastic loops. Nine children with midfacial hypoplasia were treated at their 6 - 12 years of age. No osteotomy was made in them. Bone holes were drilled with a dental bur at each side of the lateral-inferior rim of the aperture, or at the anterior part of the hard palate. The traction device was hatched to the holes through the nostrils. Protraction began 3 days postoperatively, with the forces adjusted dependently upon the rate of progress. When the skeleton reached to the planed position, it was retained with a minor force for 8 weeks.</p><p><b>RESULTS</b>The mid-facial skeleton of the nine children showed a balanced advancement. Their facial profile and cross-bite were corrected satisfactory.</p><p><b>CONCLUSIONS</b>Patients with severe mid-facial hypoplasia could be corrected ideally by the new technique, with minor trauma, easy manipulation. The design of protraction system was reasonable.</p>

Responses of dogs at different ages and long-term effects for midface skeleton protraction / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the factor of age related to protraction response.</p><p><b>METHODS</b>Four 12-week mongrel dogs in the same nest were randomly assigned into two groups: the control (n = 1) and the experimental groups (n = 3). Four pairs of titanium bone markers were fixed on either side of the bone sutures of all animals. Distraction device was fitted in the experimental group, A forward elastic force was exerted through the device for 1 month. Midface skeleton protraction was applied to experimental group at different age. Dog 2 was started at 12 weeks of age. Dog 3 was started at 16 weeks of age. Dog 4 was started at 20 weeks of age. The protraction force was 600g. All the animals were sacrificed at their age of 9 months. The results were evaluated clinically, radiographically, and cephalometrically.</p><p><b>RESULTS</b>All the animals in the experimental group showed progressively forward movement of the maxilla at the end of the experiment. In the same condition, the younger age appeared to have more effective in treatment than the older age. After 3 approximately 4 weeks stagnation, the maxilla gradually recovered normal growth.</p><p><b>CONCLUSION</b>The younger age lead more effective protraction than the older age. Persistent elastic distraction at the medium position of midface brought stable effects and no influence on facial growth.</p>

Le Fort III osteotomy and medium position distraction for mid-facial hypoplasia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To probe the possibility of distraction osteogenesis for correction of mid-facial hypplasia using a new technique.</p><p><b>METHODS</b>The distraction system of mid-facial skeleton consisted of a face-bow, the elastic loops and a device that hitched to bone holes made in the aperture rim. Flllowing Le Fort III osteotomy, a bone hole was drilled at each side of the external-inferior rim of the aperture by a dental bur. The traction device was hitched to the holes through the nostrils. Distraction began three days postoperatively, with the force adjusted dependently upon the rate of progress. When the skeleton reached to the planed position, distraction was retained with a minor force for 8 weeks. Three adults with mid-facial hypoplasia underwent this treatment.</p><p><b>RESULTS</b>The mid-facial skeleton showed a balanced advancement. The patients' facial contour and occlusal relationship recovered completely.</p><p><b>CONCLUSIONS</b>Patients with severe mid-facial hypoplasia could be corrected ideally by the new technique, with balanced advancement of mid-facial skeleton, minor trauma. The design of distraction system was reasonable. The manipulation of the procedure was easy.</p>