The polyclonal antibodies against VLDL receptor were prepared and identified. Rabbits were immunized with polypeptide fragment of VLDL receptor as antigen. The collected blood serum of the immunized rabbits was analyzed and identified by using ELISA and Western Blot. The results showed that the rabbit against mouse and human VLDL receptor antibodies were obtained with high titer and could recognize the natural VLDL receptors through Western blot. The prepared polyclonal antibodies against VLDL receptor provide a new tool to study the protein of VLDL receptor.
Antibodies/chemistry ; Antibodies/*immunology ; Enzyme-Linked Immunosorbent Assay ; Peptides/*immunology ; Receptors, LDL/*immunology

Objective To study histologic characteristics of bilayer skin substitute reconstructed by cells from human hair follicle and whether the skin appendage can be induced based on the bilayer skin substitute.Methods After composite chitosan bilayer skin substitute was reconstructed with dermal papilla cells and outer root sheath cells or dermal sheath cells and outer root sheath cells,its histologic characteristics was investigated in vitro and after transplanted onto SD albino rats.Results Composite chitosan bilayer skin substitute reconstructed by cells from hair follicle had closely arranged epithelium cells and outstanding cornification;Epithelial cords linked with epidermis could be seen in dermis.However,there was no certain hair follicle-like structure formation either in vitro or in vivo.Conclusion Hair follicle cells are good source for skin substitute reconstruction,but it can not induce skin appendage formation through skin substitute by now.

Objective T o prom ote the further research on body stature estim ation and the innovative ap-plications based on the distances betw een the anatom ical landm arks on body torso surface. Methods A specification for the collection of distances betw een the anatom ical landm arks on body torso surface w as established. T he data of 933 cases of adult population in Y angtze R iver D elta region w ere collected. M ultiple linear regression m ethod w as used to statistical analyse and establish the regression equation of stature estim ation. Results A regression equation about 5 variables including gender (x1), cervical verte-brae-coccyx line (x2), sterna-pubis line (x3), distance betw een acrom ion and iliospinale anterius (x4) and shoulder breadth (x5), and stature (y) w as established, y=105.406+5.414 x1+0.436 x2+0.286 x3+0.225 x4+0.193 x5. Conclusion T he m ethod is suitable for the rapid, sim ple and accurate estim ation of stature for the forensic experts.

OBJECTIVETO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions.

METHODA Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm.

RESULTThe calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%).

CONCLUSIONThe method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.

Astragalus membranaceus ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity

OBJECTIVETo develop a rapid and reproducible method for the culture of human fetal hair follicle bulge cells, and observe the plasticity of its differentiation into sebaceous gland in vitro.

METHODSThe bulge cells isolated from fetal human hair follicles by enzymatic digestion (digestion method) and manual microdissection (conventional method) were cultured and passaged respectively, the efficiency and biological features of cells were investigated , the clone forming efficiency was assayed by MTT, and the expression of K19 was further compared by immunocytochemistry (ABC). The morphological change and the expression of EMA of bulge cells were also observed after induction.

RESULTSBy conventional method, 8-10 bulges were harvested in one hour, 40%-50% of their cells were found to adhere to the culture plate after culturing for 48h, and they became confluent after 14 days. In comparison, about 100 bulges were harvested in one hour by digestion method, the adherence efficiency of their cells was 30% after cultivation for 12h and became confluent after 7 days. The cells grew larger with time, with irregular shape and droplets of lipid around the nucleus. The clone forming efficiency of bulge cells cultured by digestion method was (18.2 +/- 2.1) %, which was much higher than that of cells obtained by conventional method[ (12.7 +/- 3.4) %, P < 0.05]. Immunocytochemistry staining showed that positive staining of K19 was observed in most of the bulge cells, with a large amount of brown granules in the cytoplasm.

CONCLUSIONHuman hair follicle bulge cells can be efficiently cultured and multiplied in vitro, and they retained the characteristics of stem cells. And they have the potential to differentiate into sebaceous glands by induction in vitro.

Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Fetal Stem Cells ; cytology ; Hair Follicle ; cytology ; Humans ; Sebaceous Glands ; cytology

OBJECTIVETo explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (HFESCs) and the feasibility of the cultured cells as the target cells for gene transfection.

METHODSThe HFESCs were isolated by means of type IV collagen rapid adhering method. The culture medium for HFESCs was prepared according to that for human fetal fibroblasts. The cultured cells were identified by immunohistochemistry staining of keratin-19 and integrin-beta1, cell cycle analysis and clone forming rate determination. Then the cultured cells were gene transfected in vitro by liposome mediating method in which eukaryon expression vector pcDNA3.1/VEGF165 containing vascular endothelial growth factor 165 (VEGF165) were transfected into cultured cells, or by virus vector mediating method in which recombinant adenovirus accompanied vector (raav) containing green fluorescent protein (GFP) (raav/GFP) were transfected into the cultured cells, respectively. The results of in vitro gene transfection of HFESCs were observed by immunohistochemisty staining and fluorescence microscope.

RESULTSHFESCs grew well and formed large clones with higher cloning efficiency and higher ratio of G1 cells than keratinocytes. The cultured cells were strongly positive with immunohistochemistry staining of keratin-19 and integrin-beta1. After being gene-transfected by pcDNA3.1/VEGF165, the VEGF165 of HFESCs showed positive immunohistochemistry staining property, while the HFESCs transfected by raav/GFP exhibited strong fluorescence.

CONCLUSIONHFESCs could be isolated and cultured in vitro by means of rapid adherence to type IV collagen. It seemed feasible that HFESCs were gene transfected with liposome or adeno-associated virus as the vector.

Cell Adhesion ; Cell Cycle ; physiology ; Cells, Cultured ; Endothelial Growth Factors ; genetics ; metabolism ; Epidermis ; Fetus ; G1 Phase ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Keratinocytes ; cytology ; Keratins ; analysis ; Luminescent Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Microscopy, Fluorescence ; Plasmids ; genetics ; Stem Cells ; chemistry ; cytology ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

The polyclonal antibodies against VLDL receptor were prepared and identified. Rabbits were immunized with polypeptide fragment of VLDL receptor as antigen. The collected blood serum of the immunized rabbits was analyzed and identified by using ELISA and Western Blot. The results showed that the rabbit against mouse and human VLDL receptor antibodies were obtained with high titer and could recognize the natural VLDL receptors through Western blot. The prepared polyclonal antibodies against VLDL receptor provide a new tool to study the protein of VLDL receptor.
Animals ; Antibodies ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Peptides ; immunology ; Rabbits ; Receptors, LDL ; immunology

OBJECTIVETo investigate the feasibility of fabricating tissue engineering skin with human hair follicle bulge cells (HFBCs) to repair full-thickness skin wound.

METHODSHFBCs and dermal papilla cells (DPCs) isolated from human fetal hair follicles by collagenase digestion were cultured, purified and passaged. PGA-collagen scaffolds as bioengineered dermis were randomly divided into A and B groups. The HFBCs and DPCs (1 : 2) were seeded in scaffolds of group A and the equal amount of DPCs was seeded in scaffolds of group B as control. Then the keratinocyte sheets were seeded onto the surfaces of the scaffolds as bioengineered epidermis. The tissue engineering skins were then transplanted to repair the full-thickness wound on the back of nude mice. The wound healing process was observed and the plant histological changes of the transplanted engineered skin was observed with light microscope on 2, 4, 6 post-operation weeks (POW).

RESULTSThe full-thickness defect of nude mice in A and B groups could be effectively repaired by bioengineered skins. On 2 POW, integral epidermal and dermal structures were observed in the wounds in A and B groups, with thin epithelial layer and basement membrane. On 4 POW, epithelial layer became thickening and rete pegs formation was observed in basement membrane in A group, but only thickening of epithelial layer was observed in B group. On 6 POW, rete pegs structure was seen to descend and hair-follicle-like structure was formed, while only thickened epithelial layer with flat basement membrane were formed in B group.

CONCLUSIONFrom the composite skin engineered with PGA-collagen hybrid scaffolds and keratinocytes, HFBCs and DPCs could effectively repair the full-thickness skin defect of nude mice. The hair follicle stem cells participate in the process of anatomic repair of wound, and might be able to induce the repair of skin structure and function.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Dermis ; cytology ; Fetus ; cytology ; Hair Follicle ; cytology ; Humans ; Male ; Mice ; Mice, Nude ; Skin Transplantation ; Skin, Artificial ; Tissue Engineering ; methods

OBJECTIVETo study the effects qufeng zhidong Simplified Recipe (QZSR) on the behavior of the tic disorder (TD) rats.

METHODSFifty male SD rats were randomly divided to the normal group, the model group, the QZSR-1 group, the QZSR-2 group, and the QZSR group, 10 in each group. Two mg/kg apomorphine (APO) was intraperitoneally injected to rats in the model group, the QZSR-1 group, the QZSR-2 group, and the QZSR group, while equal volume of normal saline was intraperitoneally injected to rats in the normal group, both once daily for 7 successive weeks. At the 4th week equal volume of normal saline was intraperitoneally injected to rats in the model group and the normal group, while corresponding medicinal liquid was intraperitoneally injected to those in the rest groups, both once daily for 7 successive weeks. At the 2nd and 4th week of intervention, rats' improvement degrees of stereotyped behavior and the open-field test were monitored, and their experimental results were analyzed.

RESULTSAt the 2nd and 4th week of intervention, when compared with those of the model group, the score of stereotyped behavior decreased, the numbers of passing-panel, straightening, and dejecta pill were reduced, and the number of grooming increased in the QZSR-1 group, the QZSR-2 group, and the QZSR group. But there was no difference among the three groups (P>0.05).

CONCLUSIONQZSR could significantly reduce APO induced stereotyped behavior scores of TD rats, improve their locomotor activities, and reinforce their adaptive faculty.

Animals ; Behavior, Animal ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Tic Disorders ; drug therapy ; psychology

Objective To observe the stereotypy and the movement of head in rat models of tic disorder (TD) induced by apomorphine (APO) or iminodipropionitrile (IDPN),and select the best method for making rat models fully reflecting the characteristic behavior change. Methods A total of 30 male SD rats weighing 100-110 g were randomly divided into APO inducement group, IDPN inducement group and control group (n=10).The rats in the APO inducement group were intraperitoneally injected 2 mg/kg APO, and those in the IDPN inducement group were intraperitoneally injected 150 mg/kg IDPN; equal volume of saline was injected into the rats of the control group; treatments were given daily for a consecutive 7 d at a dosing volume of 1 mL/100 g. The stereotypy scale scores were observed and recorded at 5-10 min,15-20 min,25-30 min,35-40 min,45-50 min and 55-60 min after the treatment.Movement of head was noted within 5 min of sucess model making. Results The head move number of the rats in the APO and IDPN inducement groups was significantly larger than that in the control group (P＜0.05); the head move number of the rats in the IDPN inducement group was obviously larger than that in the APO inducement group (P＜0.05).The stereotyped behavior scores of rats in the APO inducement group were significantly higher than those in the control group at 5-10 min,15-20 min,25-30 min,and 35-40 min after treatment (P＜0.05); those of rats in the IDPN inducement group were higher than those in the control group at every time interval (P＜0.05); those in the IDPN inducement group were higher than those in the APO inducement group at 45-50 min and 55-60 min after tre atment (P＜0.05).Conclusion The behavioral changes of TD models induced by IDPN are more obvious than those of models induced by APO; TD model induced by IDPN is an ideal animal model for observing tic disorder.