Objective To explore the effect of botulinum toxin type A(BTXA)in the treatment of early hypertrophic scars(HTS).Methods BTXA was injected into and around the eady HTS,and then the modal and histological changes of the scars as well as the clinical reaction were observed in the patient.BTXA was also injected into muscle around the incision and effect on the cicatrization observed.Results Injection of BTXA could obviously alleviate ache and pruritus of eady HTS and could impel the atrophy and inteneration of eady HTS.Changes were found in paraffin-embedded tissue section by the hemetoxylin and eosin(HE)staining.Injection of BTXA into muscle around the cut could can reduce occurrence of HTS.Conclusion BTXA can help prevent the early HTS to a certain extent.The mechanism underlying this effect may be related to the reducing the tension around scars and proliferative activity,interfering with the signal transduction of small nerves,affecting the proliferation and apoptosis of fibroblasts and subsequently decreasing the collagen synthesis.

Objective To investigate the possible signaling mechanisms by which recombinant human platelet-derived growth factor (rhPDGF) accelerated healing of cutaneous wound in diabetic rats. Methods Four full-thickness skin wounds were incised in the back of 26 male Wistar diabetic rats. The wounded rats were divided into 3 groups (7 or 8 rats each group). One group without treatment was used as a control, and the other 2 groups were treated with rhPDGF at a dose of 7.0 μg/cm2 wound or vehicle ( DMSO/0.9%NaCl, vol/vol 1:1) from 1 to 14 days. The wound healing was evaluated by the measurements of the wound volume and area. Immunofluorescent and immunohistochemical staining were used to examine the phosphorylation of extracellular signal-regulated kinase 1/2(ERK1/2) and the expression of proliferative cell nuclear antigen (PCNA), respectively. Results Granulation tissue appeared in the bed of wound after injury. The number of blood capillary buds and fibroblasts was greater in the rhPDGF-treated group than that in the other 2 groups. A lot of inflammatory cells infiltration and collagen deposition were observed in the wound. The wound-volume in the rhPDGF-treated group was smaller than that in control group ( P ＜ 0.05). The reepithelialization rate in rhPDGF-treated group was higher than that in the other 2 groups at 7 days after injury ( P ＜ 0.05). The expression of PCNA in reparative cells was higher in rhPDGF-treated group than in control group or vehicle-treated group at 3,7 days after injury( P ＜ 0.05). The phosphorylation of ERK1/2 was stronger in rhPDGF-treated group than that in control group or vehicle group at 7 and 14 days after injury( P ＜ 0.05). Conclusion These results suggest that rhPDGF accelerates wound healing and improves healing quality by increasing the phosphorylation of ERK1/2.

Objective To investigate the dedifferentiation of epidermal cells into their progenitor stem cells induced by basic fibroblasts growth factors(bFGF) in vitro.Methods HEKa cells obtained from Cascade were found flattening and formation of cell-to-cell contacts after 6 to 7 passages,which resembled differentiated epidermal cells in vivo.To examine the effect of growth factors on the cell proliferative alterations,bFGF(100 ng/ml) was added into the culture medium for different periods(6,12,24,48,or 72 h),then the cell proliferation was measured by MTT assay.Phenotypic changes and the cell-fate determination of HEKa cells after bFGF treatment were detected by immunocytochemical assays,flow cytometry and RT-PCR analysis.HEK cells with no intervention treatment were used as a control.Results MTT assay proved that the optimal culture condition to induce the dedifferentiation of epidermal cells into their progenitors was to culture HEKa cells for 36 to 48 h when the addition of bFGF was 100 ng/ml.After treatment with bFGF for 48 h,clusters of round-shaped cells appeared around differentiated epidermal cells,and expanded progressively thereafter.These cells were smaller in shape and with larger nuclear/cytoplasm ratio,and had not only clonogenicity but also ability to form a cutaneous ridge-like structure.Immunohistochemical staining revealed that the expression levels of ?1 integrin,CK19 and CK14 were up-regulated,while the expression of CK10 was significant down-regulated after bFGF treatment.Flow cytometry indicated that there were more CK19-positive and CK14-positive cells in the treatment group than in the control(74.77% vs 15.74%,and 87.14% vs 67.26%respectively),but much lesser CK10-positive cells(4.56% vs 98.56%).Additionally,the mRNA expression levels of ?1 integrin,CK19 and CK14 were up-regulated after bFGF treatment,but that of CK10 was down-regulated.Conclusion bFGF can reverse the differentiated process of epidermal cells and induce them to produce immature,stem-like cells,which can proliferate and be used in the wound repair and regeneration of skin tissues.

Objective To investigate the optimal monochromatic parameters of CT angiography (CTA) on small feeding arteries of abdominal tumors using single source dual-energy CT with gemstone spectral imaging (GSI) technique.Methods The clinical and medical imaging data of 32 patients with abdominal malignant tumor were analyzed retrospectively during January to April 2012.Three phase-enhanced CT scans (Discovery CT750 HD,GE Heahhcare,Milwaukee,USA) of the abdomen were recorded using the GSI technique on 32 patients.The minor feeding arteries of tumor with diameter between 1.5 mm to 3.0 mm were reconstructed by 140 kVp mixed energy,66 keV,and optimal monochromatic mode respectively.After CT scanning,the original data were processed with layer and interval of 0.625 mm,the 140 kVp mixed energy images and 70 keV monochromatic images were obtained with standard algorithm.The original images were conveyed to AW4.5 work-station to process furthermore,then the data of the optimal monochromatic group and 66 keV group were reconstructed.Comparative parameters include contrast-to-noise ratio (CNR),signal-to-noise ratio (SNR) and subjective scores of the small feeding arteries CTA quality.Subjective scores were evaluated by two radiologists according to the sharpness and resolution of the small feeding arteries.One-way ANOVA was used to for statistical analysis.Results CNR of the optimal monochromatic group,the 66 keY group,and the 140 kVp mixed energy group were 21.70 ±9.74,16.63 ±7.60,and 9.85± 6.76,respectively.SNR were 35.05 ± 17.75,26.77 ± 11.51,and 16.32 ± 9.5,respectively.Subjective scores were 4.58 ± 0.40,3.55 ± 0.57,and 2.75 ± 0.46,respectively.CNR,SNR and subjective scores had significant difference among groups (F =17.11,15.73 and 116.01,P ＜ 0.01).The optimal monochromatic group was superior to the 66 keV group and the 140 kVp mixed energy group.Conclusion The optimal monochromatic mode can improve CTA quality of small feeding arteries of abdomen malignant tumors with GSI technique using single source dual-energy CT.

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.

BACKGROUND:There is no literature about the treatment according to the mechanism of cervical spine injury classification, especial y for the treatment of extension type cervical fracture/dislocation with merger cases of posterior composite structure damage, whether simple anterior approach can meet the needs of the treatment has no detailed elaboration. This article may analyze from the aspect of cervical spine injury mechanism. OBJECTIVE:To observe the short-term effect of anterior cervical Cage-assisted fusion combined with locking titanium plate internal fixation for the treatment of extension type cervical fracture. METHODS:A retrospective analysis was performed in 15 extension type cervical spine fracture dislocation patients treated with decompression anterior cervical intervertebral disc resection plus bone graft with cage-fusion locking titanium plate internal fixation from June 2006 to March 2011 in the Department of Orthopedics, Xianning Central Hospital, including 10 cases of single segment injury and treatment, and five cases of multiple segment injury and treatment. Japanese Orthopaedic Association score and the neck disability index were compared before and after treatment;the cervical flexion and height were measured according to the antersposterior X-ray film taken before fixation, 1 week after fixation and final fol ow-up.RESULTS AND CONCLUSION:The patients were fol owed-up for 8-37 months. One case had Cage mild sinking and shift, and there was no internal fixation breakage or loosening in al the patients. Transient pharyngeal discomfort was observed in 11 patients. Compared with the preoperation, the Japanese Orthopaedic Association score, neck dysfunction index, fusion segment cervical flexion and fusion segment intervertebral disc height were significantly improved at 1 week after fixation and final fol ow-up (P<0.05). There were no significant differences between 1 week after fixation and final fol ow-up (P>0.05). The short-term effect of decompression anterior cervical intervertebral disc resection plus bone graft with cage-fusion locking titanium plate internal fixation for the treatment of extension-type cervical fracture is good.

AIM: To investigate the mechanism of proliferation and differentiation of human epidermal stem cells cultured in vitro under the influence of compressive stress.METHODS: Epidermal stem cells were isolated by adhering to type IV collagen and were cultured with conditioned medium,then were detected by Powervision~(TM) two-step immunohistochemical method with keratin 19 and cell cycle analysis.The cultured epidermal stem cells transplanted on silica gel membranes,which were put in a new apparatus,was designed to offer cell culture and intermittent compressive stress(4 kPa,6 kPa,8 kPa,10 kPa,12 kPa) for 2 h,3 times a day simultaneously.A week later,cells on silica gel membranes were identified with keratin 19 and 10 by Powervision~(TM) two-step immunohistochemical method.RESULTS: The new apparatus offered cell culture and intermittent compressive stress simultaneously.The isolated and cultured epidermal stem cells were identified with keratin 19 positive and 84.80 percent of them were showed in G_1 period with cell cycle analysis.Cells on silica gel membranes had been subjected intermittent compressive stress above 8 kPa for a week.The number of the cells was increased,which was more than that in control group.However,some cells identified by immunohistochemical staining with keratin 10 positive were detected among the disposed epidermal stem cells.CONCLUSION: The intermittent compressive stress above 8 kPa induces and promotes epidermal stem cells to proliferate and differentiate,indicating that epidermal stem cells respond to mechanical stress,probably is one of their major biological features.

BACKGROUND:Chinese herb Cape Jasmine Fruit can activate the chondrocytes and geniposide is an important component in this herb.
OBJECTIVE:To investigate the effect of geniposide on col agen II synthesis in rat chondrocytes cultured in vitro. METHODS:Rat chondrocytes were separated and cultured in vitro. The chondrocytes were then interfered with 25, 50 and 100 mmol/L geniposide. Normal control group was also set. Col agen II mRNA and protein expression was detected with semi-quantitative RT-PCR and western blot analysis, respectively.
RESULTS AND CONCLUSION:RT-PCR and western blot analysis results showed that, geniposide at 25, 50 and 100 mmol/L increased the col agen II mRNA and protein expression (P<0.01). Geniposide can promote the synthesis of col agen II in rat chondrocytes cultured in vitro.