0 05). The serum levels of both IL-8 and TNF (P

Objective: To search for a simple and rapid cell culture method for human gingival epithelial cells with a high success rate. Methods:Culture medium containing serum has been proved to have the ability of accelerating the early adhesion of human gingival tissue blocks, and the migration of gingival epithelial cells from the rim of the blocks. By means of this, we introduced the serum containing DMEM to the cell culture medium within the first 7-10 days, and changed with serum free cell culture medium to accelerate the mitosis, proliferation, and migration of the gingival epithelial cells, which had moved out from the tissue blocks. Using the combined method, cells were identified by the method of morphology, immunohistochemistry and analysis of the cell growth curve, as well as SEM. Compared study of the effects on the cell growth between combined method and serum containing DMEM or serum free EpiLife culture method was conducted. Results: Cells were harvested within 17-22 days. Primary culture success rate was 90.6%. Cultured cells were slabstone shaped. Immunohistochemistry analysis of keratin antibody showed a positive result. The cells had stronger ability of migration and proliferation in the serum free cell culture medium compared with that in the serum medium. Conclusion: This method can culture the gingival epithelium cells conveniently with accelerated speed.

Objective: To evaluate the possibility of self-assembly oligopeptide(T2) for dental enamel biomimetics, especially for the prism’s crystal texture since it could prompt calcium phosphate precipitated in gel carrier. Methods:SEM (Scaning electron microscope) and TEM (Transmission electron microscope) were used to observe the morphologic presentation and ED(Electron diffraction) to crystal texture comparing with the human molar enamel powder. Results: (a) Flake-like and needle-like octacalcium phosphate precipitated in the gel carrier with self-assemble oligopeptide(T2). They transformed into rod-like hydroxyapatite crystals gradually in the following 2-4 weeks. (b) The rod-like hydroxyapatite may arrange or grow into bundles which are similar to the human enamel prisms in both appearance and size. (c) The rod-like hydroxyapatite showed polycrystal while the enamel prisms showed monocrystal under examination of ED. Conclusion:The self-assemble oligopeptide(T2) could regulate the speed of nucleation and crystallization of hydroxyapatite in morphology and crystalline size. Thus, the self-assembly oligopeptide and the gel carrier mineralization system could be primarily applied in biomimetic use for the crystallization of hydroxyaptite in dental prism in vitro.

OBJECTIVETo establish a new method for evaluating quality stability of Shudihuang decoction pieces with near-infrared spectrum.

METHODClustering analysis was used to distinguish different Shudihuang samples. And comparisons between NIR and HPLC fringerprint.

RESULTResults of clustering reflected the similar degree among these samples. Compared with HPLC fingerprint, NIR analytical process fast and convenient with exact results.

CONCLUSIONNIR is a feasible and effective approach to examine the quality stability of Shudihuang decoction pieces with unique virtues.

Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; standards

Aged ; Aged, 80 and over ; Antibiotics, Antitubercular ; pharmacology ; Coal Mining ; DNA Gyrase ; genetics ; Drug Resistance, Bacterial ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; Pneumoconiosis ; complications ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Quinolones ; pharmacology ; Tuberculosis, Pulmonary ; complications ; microbiology

HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.

Objective·To construct C-shaped cartilage rings by rabbit auricular cartilage-derived chondrocytes combing with both electrospun gelatin/ polycaprolactone(GT/PCL) nanofibrous membranes and 3D printed supporters for repairing tracheal cartilage defects.Methods·Primary chondrocytes were isolated from rabbit auricular cartilage with methods of trypsin enzyme digestion and collagenase enzyme digestion.After proliferation in vitro,the chondrocytes of passage 2 were harvested for further experiments.Ultrafine composite fibers of GT/PCL were fabricated via electrospinning.The electrospun GT/PCL membranes were tailored into rectangle shape,the length of which is 12 cm and the width is 2.5 cm.Chondrocytes were seeded on membrane at a density of 1 × 108 cells/mL.Then the membrane were rolled onto a 3D printed supporter of poly(DL-lactide-ε-caprolactone) (PLCL) material to construct a C-shaped cartilage-like complex.After 8 weeks of subcutaneous incubation in vivo,gross inspection and paraffin section staining were applied for evaluation.Results·After 8 weeks of culture in vivo,mature cartilage-like tissue were formed with open-cylindrical bellow appearance and pecific mechanical property.C-shaped rings arranged at regular intervals on the inner surface of tissue,which were similar to the normal structure of tracheal cartilages.Histological and immunohistological staining showed a large number of typical lacunar structures and extracellular matrix secretions.Conclusion·It is feasible to construct tissue engineered C-shaped cartilage tissue by combing chondrocytes with GT/PCL membrane and 3D printed PLCL supporter for tracheal cartilage repair.

Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; Glycyrrhetinic Acid/pharmacology* ; Humans

OBJECTIVE: To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle mechanical injury in rats. METHODS: The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury) and control group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. RESULTS: At post-injury 6-12h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN decreased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14d, the percentage of FBC reached peak. CONCLUSION The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.
Animals ; Fibroblasts ; Muscle, Skeletal/injuries* ; Neutrophils ; Rats ; Time Factors

OBJECTIVETo investigate the effect of pulsed electromagnetic fields (PEMs) on inducing osteoclastic like cell (OLC) formation changes and apoptosis in ovariectomized (OVX) rats bone marrow culture in vitro.

METHODSThirty healthy three-month-old female Wistar rats were either sham-operated (Sham) or ovariectomized (OVX) and randomly divided into three groups: group A (OVX + PEMs, 18 rats), group B (OVX, 6 rats) and group C (Sham, 6 rats); group A was again randomly divided into three groups: A1, A2, A3. The frequencies adopted were 1.5, 2, 75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months, then the bone marrow of all rats were cultured, 2 days later, group A cells (including group A1, A2, A3) were collected and exposed to different frequencies PEMs for 2 weeks (30 min/day). In order to observe the changes of osteoclasts and count their numbers, cells were taken for Wright Giemsa staining, tartrate-resistance acid phosphatase (TRAP) staining and Hoechst 33258 staining.

RESULTSTRAP staining results indicated the number of OLC in group C was the least, then was group A2, A3, A1, B. The number of OLC in group B was remarkably increased (P < 0.01; vs group C, A2). The number of OLC in group B was significantly increased (P < 0.05; vs group A1, A3). Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups, which of group C, A2 was significantly increased (P < 0.05; vs group B).

CONCLUSIONPEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized (OVX) rats bone marrow culture in vitro, the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy.

Acid Phosphatase ; metabolism ; Animals ; Apoptosis ; radiation effects ; Bone Marrow Cells ; cytology ; enzymology ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Female ; Isoenzymes ; metabolism ; Osteoclasts ; cytology ; enzymology ; radiation effects ; Ovariectomy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase