Objective To explore the isolation,amplification methods and adipogenic differentiation under specific culture medium of human keloid-derived precursor cell (KPC) in vitro,in order to study their possibility of being new seed cells of tissue engineering fat.Methods KPCs were isolated from human keloid tissue of 4 different patients in our hospital and were cultured in the modified L-DMEM culture medium.Their cloning efficiency and growth curve were tested.The subcultured cells were tested of the mesenchymal stem cell (MSC)-related gene expression by flow cytometry.In addition,they were cultured in H-DMEM medium (containing 1 μmol/L dexamethasone,0.5 mmol/L 3-isobutyl-1-methyl-xanthine 10 mg/L of bovine insulin,100 mmol/L indomethacin,and 10 ％ FBS)and were later observed in oil red O staining under phase contrast microscope to determine whether lipid droplets generation was formed,using skin-derived precursors (SKP) as control.Results More than 95 ％ KPC expressed many antigens of MSC,such as CD29,CD44,CD90 and CD105 while few of them expressed CD34,CD45(1.0 ％-2.5 ％).And the cells increased in size gradually after inducted the same time,changing from spindle into round or polygonal in shape.The lipid droplets were seen in 72 hours and expressed a positive rate of 78.6 ％ in Day 19 in oil red O staining while the same rate was 54.6 ％ in SKP.Conclusions Human keloid-derived precursor cells can express a variety of MSC-related surface markers without expressing hematopoietic stem cell (HSC) related markers.Furthermore,they can be differentiated into fat cells under certain conditions,which may make them as a new source of seed cells for tissue engineering fat.

OBJECTIVEDehydroandrographolide (DP) from Andrographis paniculata (Burm. F.) Nees is a potential anticancer agent. This study aimed to investigate the oral bioavailability and intestinal disposition of DP to provide useful information for the development of DP as a new candidate anticancer drug.

METHODSThe pharmacokinetics of DP was evaluated in rats, and its intestinal disposition was determined using cultured Caco-2 cells and a single-pass rat intestinal perfusion model.

RESULTSThe oral bioavailability of DP was 11.92% in rats. The apparent permeability coefficient (P(app)) of DP from the basolateral side (B) to the apical side (A) (5.37×10(-5) cm/s) of the Caco-2 model was roughly equal to that from A to B (4.56×10(-5) cm/s), suggesting no involvement of the efflux transporter(s). In the perfusion model, no significant difference was found in the effective permeability (P*(eff)) of DP between the 4 segments of the intestine. No significant metabolism of DP was detected in the intestinal perfusates. The amount of DP found in the bile was only about 0.1% of the absorbed amount. The P*(eff) and bile amounts of DP were not significantly increased by P-glycoprotein (P-gp) inhibitor or breast cancer resistant protein (BCRP) inhibitor (P>0.05).

CONCLUSIONThe bioavailability of DP was 11.92% in rats. DP has good absorption and metabolism stability in the intestine. The efflux transporters such as P-gp and BCRP do not participate in DP transport.

Administration, Oral ; Animals ; Biological Availability ; Biological Transport ; Caco-2 Cells ; Diterpenes ; administration & dosage ; pharmacokinetics ; Humans ; Intestinal Absorption ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To evaluate the clinical efficacy of the bridge pedicled transplantation of medial leg fascial flap combined with medial hemisoleus muscle flap for contralateral leg soft tissue defect.Methods:Between January of 2012 and January of 2018, 12 patients with soft tissue defect of the leg were treated with bridge pedicled transplantation of contralateral medial leg fascial flap combined with medial hemisoleus muscle flap by posterior tibial artery. There were 9 males and 3 female, aged from 19 to 53 years (mean, 35 years). The size of the soft-tissue defects ranged from 12 cm×8 cm to 18 cm×9 cm. The immediate coverage of the fascial and muscle flaps and vessel pedicle were repaired by a meshed split-thickness skin graft. The donor site was closed directly. After the transplantation of the one pedicle and two flaps survived, vascular pedicle was cut off.Results:All the fascial and muscle flaps survived completely. No clinical vascular deficiency was found on the fascial and muscle flaps postoperatively. One case developed distal muscle flap small skin graft necrosis, and spontaneous healed after 2 weeks of dressing change. Follow-up period ranged from 2.5 to 4.5 years (mean 3.8 years). A good contour was confirmed both at the recipient and donor sites. Satisfactory clinical results were obtained in this series.Conclusion:This method is suitable for the treatment of soft tissue defects of the leg with only one major blood vessel, which reduces the damage to the donor site.

Objective:To evaluate the efficacy of the repairing anterior tibial adjacent double wounds by transposition with single-pedicle two flaps of the medial head of gastrocnemius.Methods:Between January of 2012 and January of 2018, 10 patients with the anterior tibial adjacent double wounds (7 males and 3 female, aged from 21 to 45 years) were treated by transposition with single-pedicle two flaps of the medial head of gastrocnemius. The size of the soft-tissue defects ranged from 2.0 cm×2.5 cm to 4.5 cm×4.0 cm. The medial head of the gastrocnemius was divided into two flaps with a single pedicle to repair two adjacent wounds of the anterior tibial. The muscle flaps were immediately covered by a meshed split-thickness skin graft, and the wound in donor site was closed directly.Results:All the muscle flaps survived completely. No clinical vascular deficiency was found on the muscle flaps postoperatively. Small wound dehiscence was developed in one patient and spontaneously healed 2 weeks after dressing change. Patients were followed up for 2.0 to 4.5 years. A good contour was confirmed at the recipient and donor sites. Satisfactory clinical results were obtained in this series.Conclusion:This method is suitable for the repair of two adjacent small wounds of the anterior tibial which can reduce the damage to the donor site.

Objective:To evaluate the pedicled bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap for the treatment of soft tissue defects at the contrallateral leg.Methods:Between January of 2012 and January of 2016, 8 patients with soft tissue defects at the leg were treated at Department of Orthopedic Surgery, Hand and Foot Surgery Hospital of Lanzhou. They were 5 men and 3 women, aged from 19 to 50 years (mean, 35 years). All of them were treated by bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap pedicled with posterior tibial artery. The size of the defects ranged from 10 cm×9 cm to 13 cm×8 cm. The immediate coverage of the muscle flaps and vessel pedicle was repaired by a meshed split-thickness skin graft. The donor site was closed directly. The therapeutic efficacy was assessed at the final follow-up according to the criteria by Iowa for tibial fractures.Results:All the skin flaps and muscle flaps survived without any vascular crisis. One case developed necrosis of small skin graft at the distal muscle flap which spontaneously healed after dressing change for 2 weeks. Their follow-up ranged from 2.5 to 4.5 years (mean, 3.8 years). A good contour was confirmed at the recipient area. By the Iowa criteria at the final follow-up, 3 cases were excellent, 4 good and one fair.Conclusion:Pedicled bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap is a good treatment for soft tissue defects at the contrallateral leg which has only one major blood vessel, reducing damage to the donor site.

Objective The contents of 11 nucleosides and base components in 10 batches of samples from 5 provinces(cities)including Chongqing,Yunnan and Shaanxi were determined,and the differences in nucleosides and base components in Fritillaria taipaiensis were compared by chemometric analysis,and the quality was comprehensively evaluated,so as to provide a reference for the cultivation of excellent varieties and the selection of medicinal materials.Methods Nucleoside and base components were extracted from Fritillaria taipaiensis by ultrasonication in aqueous solutions,and the content of each component was determined by HPLC-DAD method.The origin was classified by principal component analysis(PCA)and hierarchical cluster analysis(HCA).Partial least squares discriminant analysis(PLS-DA)was used to determine the differentiated index components in Fritillaria taipaiensis.Then the differences in the contents of the index components among samples from different origins were compared.Results It was found that 11 nucleoside and base components differed significantly among different origins of Fritillaria taipaiensis.Principal component analysis and hierarchical cluster analysis indicated that all samples could be clustered into 4 categories.Five characteristic components,including uracil,cytosine,uridine,inosine,and adenosine,were identified by PLS-DA.The nucleosides and bases in samples from Chongqing and Hubei were relatively high,and the quality of the samples was comparatively superior.Conclusion This method is simple,reproducible,accurate and reliable.It has screened out the index nucleoside and base components in the identification of Fritillaria taipaiensis of different origins,which can be used to initially elucidate the differences of samples between different origins.Additionally,it can better reflect the quality of Fritillaria taipaiensis,and can provide reference for the selection of procurement origin and the quality control for Fritillaria taipaiensis.