BACKGROUND: Up to date, studies concerning capspase 3 inhibitor mainly focus on peptide/non-peptide compounds synthesis and detection. Few reports addressing inhibits chondrocytes apoptosis using silenced caspase 3 gene. OBJECTIVE: To inhibit apoptosis of chondrocytes by blocking the apoptotic cascade reaction, gene silencing of caspase 3, and transduction of caspase 3 siRNA into chondrocytes with lentivirus vector.DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Traumatic Surgery, Guangzhou Red Cross Hospital, Medical College of Jinan University from June 2008 to June 2009.MATERIALS: Chondrocytes were harvested from SD rats, and caspase 3 shRNA plasimid was constructed by our laboratory.METHODS: Rattus caspase 3 siRNA was synthesized and cloned into pSIH1-H1-copGFP plasmid. pSIH1-H1-copGFP-caspase 3 siRNA lentivirus was generated in 293TN cells by pPACKH1~(TM) Lentivector Packaging Kit and transducted into chondrocytes of rats.MAIN OUTCOME MEASURES: After the lentivirus was transducted into chondrocytes, the caspase 3 mRNA was tested by RT-PCR and the caspase 3 protein was tested by Western blot. Both the transducted cells and untransducted cells were induced apoptosis by tumor necrosis factor α (TNF-α). Cell apoptosis was assessed by flow cytometry, Annevin V/PI.RESULTS: The transduction rate of caspase 3 siRNA was about 90% by lentivirus vector. The expression of caspase 3 mRNA and caspase 3 protein in transducted chondrocytes was lower than the normal chondrocytes (P < 0.01). When the cells induced apoptosis by TNF-α, the apoptosis rate of the negative siRNA- chondrocytes was 7 times higher than that of caspase 3 siRNA-chondrocytes.CONCLUSION: The caspase 3 siRNA could inhibit caspase 3 expression and decrease drug-induced apoptosis of the chondrocytes.

Objective The purpose of the present study is to illustrate the effect of olfactory ensheathing cells(OECs) on the survival and neurite outgrowth of GABAergic neurons in vitro. Methods OECs were dissociated from olfactory bulb and neurons from spinal cord of E12 mouse. On the sixth day in vitro,the Millipore cultue blank with OECs was transferred to the neuron culture mediam and continue the co-culture for another 6 days.The cultured neurons were stained with anti-GABA antibody.The neurite of neurons was observed with an image system.The number of GABAergic positive neurons was counted under the microscope. Result The number of GABAergic neurons was 39^7?6^3 in co-culture groups,whereas the number of GABAergic neurons represented only 27^6?2^7 in control groups(CG),(P

BACKGROUND: Simple polyvinyl alcohol (PVA) has limited ability to cell adhesion. There are not generally accepted studies on improved effects of collagen protein modified polyvinyl alcohol on cell adhesion and proliferation.OBJECTIVE: To investigate the feasibility of PVA/type Ⅰ college (COL-Ⅰ) as anterior cruciate ligament (ACL) scaffolds in tissue engineering.DESIGN, TIME AND SETTING: The controlled observation experiment was performed at the Fourth Affiliated Hospital, Medical College. Ji'nan University, Guangzhou Red Cross Hospital, Guangzhou Institute of Trauma Surgery from August 2006 to October 2007.MATERIALS: COL-Ⅰ gel was produced by Guangzhou Institute of Trauma Surgery.METHODS: PVA filature was used to weave fascicular scaffolds. NIH-3T3 cell line and human ACL cells were in vitro incubated, amplified, and then implanted on the PVA/COL scaffolds.MAIN OUTCOME MEASURES: The growth of NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds and the secretion of extracellular matrix were observed using scanning electron microscope. Cell compatibility of PVA/COL scaffolds was assessed. Mechanics characteristic of PVA/COL scaffolds was measured by using the electric. tensile force apparatus. Mechanical property of PVA/COL scaffolds was analyzed using the SPSS 11.5 software package.RESULTS: NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds adhered, proliferated, and secreted extracellular matrix. NIH-3T3 cell line highly grew compared with human ACL cells on the PVA/COL scaffolds. The adhered number of NIH-3T3 cell line and human ACL cells was significantly increased on the PVA/COL scaffolds. NIH-3T3 cell line and human ACL cells presented well morphology on the PVA/COL scaffolds. COL-Ⅰ could promote the secretion of extracellular matrix from NIH-3T3 cells, but its effects on human ACL cells were not significant. Tensile force test showed that load-extension curve of the materials was identical to ACL of human and rabbits, and the scaffolds possessed strong flexibility. The maximal load, ultimate stress and elastic modulus were respectively 52.61 N, 14.96 MPa and 202.08 MPa.CONCLUSION: COL-Ⅰ accelerates the adhesion and proliferation of NIH-3T3 cell line and human ACL cells on the surface and in the pore of the PVA/COL scaffolds, promotes the secretion of extracellular matrix from NIH-3T3, and PVA filature material has mechanical property and good cell compatibility.

Objective To evaluate the immuno-potentiating effects of CpG-ODN plus alum as a composite adjuvant on influenza split virion vaccine.Methods BALB/c mice were immunized with various amounts of 2009 H1N1 influenza split virion vaccine,alone or in combination with CpG-ODN,alum,or both (composite adjuvant).Antigen-specific humoral immune responses were evaluated by ELISA,hemagglutination inhibiting (HI) assay and neutralizing assay.Antigen-specific cellular immune responses were evaluated by ELISPOT assay,intracellular cytokine staining assay and in vivo CTL assay.Results Compared with the control group immunized with antigen alone,a single use of either adjuvant weakly enhanced the humoral immune responses,as indicated by the increase of antigen-specific IgG titers,HI titers and neutralizing titers by 3-6 folds,2-4 folds and 4-8 folds,respectively,after two immunizations.In contrast,the composite adjuvant induced more potent humoral immune responses; the antigen-specific IgG titers,HI titers and neutralizing titers were increased by 23-57 folds,9-20 folds and 16-64 folds,respectively.Consequently,the composite adjuvant achieved antigen-sparing by at least 16 folds.In addition,the composite adjuvant significantly enhanced the antigen-specific cellular immune responses,as revealed by the increase of IFN-γ-secreting CD4+ T cells and the enhancement of CTL activity in immunized mice.Conclusion CpG-ODN plus alum as a composite adjuvant can enhance the immunogenicity of influenza split virion vaccine and achieve the antigen-sparing effect.

Objective To conduct intensive clinical skill training for non-mainland medicine interns and to explore the effects of training.Methods Twenty-six new clinical interns in the first affiliated hospital of Jinan university from June 2019 to June 2011 were enrolled and the 96 hours of intensive training was conducted.Results The scores of theoretical examination,puncture operation,physical examination and complete medical records of the 26 non-mainland interns were improved after training ( P ＜ 0.05 ) and were higher than those of mainland interns ( P ＜ 0.05 ).The enhancement in complete medical recording skills was the greatest and in puncture operation was the smallest among the four techniques.Conclusions The intensive training can effectively improve the clinical skills of nonmainland medical interns and the improvement of clinical skills in non-mainland interns is greater than that in mainland students.

BACKGROUND: The repair of articular cartilage defect is always a problem that is dedicatedly solved by doctors of orthopaedics. Autologous perichondrium, periosteum or allografting of cartilage have been used previously; however, the source of the donor is limited, the fixation is difficult as well as the occurrence of endochondrial ossification and delamination between the inferior cartilage and reparative cartilage, etc. Type Ⅱ collagen, the main component of cartilage matrix, has certain effects in the repair of articular cartilage defect.OBJECTIVE: To investigate the effects of type Ⅱ collagen sponge filling on the repair of articular cartilage defect.DESIGN: A randomized controlled trial.SETTING and MATERIALS: The study was conducted in the Guangzhou Institute of Traumatic Surgery. Materials were 24 adult male purebred New Zealand Rabbits(48 knees), ordinary grade, with a body mass of (2.29 ±0. 25) kg. Animals were fed with standard feeding in separate cage.INTERVENTIONS: A full-thickness defect in articular cartilage was made on the femoral trochlear surface by a drill of 5 mm in diameter and 3 mm in depth. Rabbits were allocated into filling group(type Ⅱ collagen sponge was grafted into left keen joint defect) and control group(right knee joint defect site was set as control) according to random number table.MAIN OUTCOME MEASURES: Gross morphological and histological observation of the defect repair in each dual week within 12 weeks after operation.RESULTS: During 10 - 12 weeks, in cuntrol group: The defect area was repaired by white and soft tissue that had no resistance to press. The repaired tissue was still lower than the surrounding articular surface with clear boundary. By histological observation, it was found that the defect was repaired by the mechanism similar to inflammatory reaction and the defect is ultimately filled by the hyperplasia of hyaline degenerative fibrous tissues. In filling group: the defect was repaired by semi-transparent, smooth, textured tissues with polish that had resistance to press as well as elasticity. The repaired tissue was almost similar to the shape of the surrounding cartilage,difficult to be distinguished. After histological observation, it was found that there was no inflammatory reaction, but active hyperplasia of inner bonetissue and cartilage tissues; a lot of osteoid tissues and trabeculation were found. Newlborn cartilage was fused with surrounding cartilage tissue and connected with surrounding tissues. Type Ⅱ collagen had significantly promoting effects in the repair of articular cartilage defect, and the repaired cartilage was close to normal cartilage. CONCLUSION: The self-made high purity type Ⅱ collagen sponge has favorable promoting effects in the repair of articular cartilage defect with good histocompatibility but without obvious toxic side effect.

Objective To improve the teaching methods of graduate students, and provide the theoretical basis for other teaching hospitals to extend the preclinical training mode of clinical medicine professional degree students. Methods 90 clinical medical postgraduate students of Grade 2012 were divided into four groups according to their specialized field and 54 hours of pre-job clinical intensive training were conducted at the same time. The training included four modules lectured by teachers with physician-in-charge above title, such as communication between doctors and patients and medical history collection, physical examination, medical record writing as well as theory of knowl-edge. Before and after the implementation of intensive training, these four skills and knowledge of the students were tested and assessed by professional teachers. Relevant data were paired t test, ANOVA and non-parametric tests. Results The test results of patient-doctor communication and patient history collection, physical examination, complete medical records and theoretical examination results of each group after training were improved to be higher than before (P<0.05). And the effect of patient-doctor communication and patient history collection was the most significant. The doctor-patient communica-tion and history acquisition performance of different groups of graduate students had statistically sig-nificant difference (P=0.001). Conclusion Strengthening the preclinical comprehensive training for medical postgraduate can make the students master the basic clinical skills better and more effectively and the corresponding training methods are worthy of popularization to the other teaching hospitals.

Objective To compare the growth property of the stem cells taken from different brain regions at the same developmental stage. Methods Mice embryos at the same development stage were isolated under sterile conditions, cortex, striatum, diencephalon, mid-hind brain and spinal cord were collected and pooled separately, after single-cell suspension obtained, different regions' cell suspensions were seeded in FGF supplemented serum-free culture medium. Followed the neural stem cell clone(neurospheres) fromation, immuno-cytochemistry method was utilized to identify the cell characteristics, all these clones were passaged under same conditions, clone formation and cell migration were observed under phase-contrast microscope. Results In the FGF added serum-free medium, neural cells experienced a large scale death within 48h after being seeded, then few single cells began to proliferate and formed the floating cell clones in the medium. These clones (neurospheres) could form new clones when seeded as single cell suspension. If these clones were seeded on poly-orithine, they could differentiate into neurons and glia cells. Compare the clone formation and cell migration, we found that: cortex, striatum, diencephalon all could form floating clones with different rate, the cortex formed clones at the highest rate, striatum and diencephalon at lower rate; few neurospheres formed from cortex adhered to the culture plate substrate and few cells were found migrating out from the adhered clones, striatum and diencephalon derived neurospheres adhered the plate more easily, and there were apparent cell migration. Mid-hind brain and spinal cord formed clones at the lowest rate, floating clones were scarce, and the clones adhered to the substrate readily, there were large amount of cell migrating out from these adhered clones. Conclusion Neural stem cells could proliferate and be passaged in vitro in serum-free medium, and they could be induced to differentiate under certain conditions into major cells types of CNS, there were differences in clone forming rate and cell migration between neural stem cells derived from different CNC regions, nonetheless they were at the same development stage, this may reflect that, in some degree, these cells can keep some of their region-specified developmental intrinsic property in vitro.

Objective To evaluate the efficacy of 1-stage urethroplasty using pedicle circular fascioctaneous preputial flap for the treatment of complex anterior urthral strictures.Methods Between January 2006 and January 2013, 37 patients with complex anterior urethral stricture were treated by 1-stage urethroplasty using pedicle circular fascioctaneous preputial flap.The mean age was 41 years ( 22 -71 years) .The etiology of stricture included trauma of 13 cases, iatrogenieity of 13 cases, gonorrhea infection of 2 cases, unknown reason of 9 cases.The penile urethral stricture was found in 22 cases, the bulbourethral stricture in 9 cases, and stricture extending from penile to posterior urethra in 6 cases.The mean length of anterior urethral stricture was 8.1 cm (range 5.0-14.0 cm).A circumferential island of the preputial/distal penile skin was mobilized by the technique of preserving penile fasciocutaneous wide vascular pedicle. The pedicle is composed of two layers of the dartos and the superficial lamella of Buck′s fascia, and the flap was divided in the midventral/middorsal plane back to the penoscrotal junction to convert the circular configuration to a longitudinal trip for urethral reconstruction.The dorsal and ventral inlaid flap urethroplasty was performed in 27 cases and tubularized flap urethroplasty was performed in 10 cases.Results The mean operative duration was 3.1 h (2.5-3.5 h).The mean length of the circular fascioctaneous preputial flap was 10.4 cm (range 9.0 -14.0 cm).All the patients were followed up for mean 22 months (3 -51 months).Thirty-two cases voided well and the mean peak urinary flow rate was 22.3 ml/s (15.0-29.0 ml/s).One-stage healing achieved in 32 cases (86.5%).Recurrent stenosis was noted in 4 cases, and meatal stenosis occurred in 1 patient, who required re-operation.Re-repair succeeded in 4 cases and total success rate was 97.3% (36/37).Conclusions The pedicle circular fascioctaneous preputial flap has advantage of good blood supply and autograft for new meatus.It could be a reliable and durable method for the treatment of complex anterior urthral strictures(≥5 cm) in 1-stage urethroplasty.

The preparation of collagen sponges was studied in order to develop tissue engineering scaffolds. Collagen solutions with varying concentrations were obtained by condensing the initial collagen with polyethylene glycol (PEG) at 4 degrees C for different periods of time, and then were freeze-dried to make collagen scaffolds. The porous characteristics of the prepared scaffolds were characterized by use of different methods, including laser scanning confocal microscopy (LSCM), scanning electron microscopy (SEM) and tensile tests. All collagen sponges were shown to have similar interconnected porous structures but were found to have different pore size, porosity, water capacity and the mechanical property, depending on the concentration of collagen solutions. These findings indicate that the way of controlling the concentration of collagen solutions with PEG permits the freeze-drying fabrication of collagen sponges with varying porous features suitable for different tissue engineering purposes.
Collagen ; chemistry ; ultrastructure ; Freeze Drying ; Polyethylene Glycols ; chemistry ; Porosity ; Tissue Engineering ; Tissue Scaffolds