BACKGROUND: There exists insufficient study with specific applicability regarding vascular distribution characteristics of sural nerve nutrient vessels of the distally based flap.OBJECTIVE: To investigate the distribution of sural nerve nutrient vessels of the distally based flap and provide an anatomical evidence for the design of operation on repair of foot injury.DESIGN: A single sample study.SETTING: Research Center of Clinical Anatomy, Fuzhou General Military Hospital of Nanjing Military Area Command of Chinese PLA, and Department of Orthopaedics, the 97 Hospital of Chinese PLA.PARTICIPANTS: Thirty-two samples of lower extremities whose blood vessels were perfused with red emulsion were provided by Research Center of Clinical Anatomy, Fuzhou General Military Hospital of Nanjing Military Area Command of Chinese PLA.METHODS: The origin of the blood vessels of distally based flap and deep communicating branches of the lesser saphenous vein in the samples were dissected and observed.MAIN OUTCOME MEASURES: ①The nutrient vessels of sural nerve of distally basde flap.②The nutrient vessels of lesser saphenous vein of distally based flap.③The superficial and deep communicating branches of lesser saphenous vein of distally based flap.RESULTS: There were 2 to 5 nutrient vessels in the distally based flap:one originating from the perforating branch of the lateral calcaneal artery was (0. 6 ± 0.2) mm in diameter and one from the terminal perforating branch of the peroneal artery was(0.8±0. 2) mm in diameter and they were (1.0 ± 1.3) and(2. 8 ± 1.0) cm, respectively, away from the lateral malleolus. The incidence of intermuscular septum perforating branches (0 to 3) was 96. 7%, 66.7% and 20. 0%, respectively, and their diameter was (0.9 ±0. 3), (1.0 ±0. 2) and (0. 8 ±0. 4) mm and their distance to lateral malleolus was(5.3 ±2. 1), (6. 8 ±2.8) and (7.0 ±4.0) cm, respectively. There were 2 types of nutrient vessels of the lesser saphenous vein of distally based flap, the nutrient vessels of nerve-vein and the ones of vein-nerve. The superficial and deep communicating branches of the lesser saphenous vein were(1.7 ±0. 5) mm in diameter and(3.4 ±0. 9) cm away from the lateral malleolus. They ended at the peroneal veins.CONCLUSION: The perforating branches of heel lateral artery, the terminal perforating branches and intermuscular septum perforating branches of the peroneal artery have sub-branches to deep fascia, skin, nerves and parenteral nutrient vessels. These sub-branches communicate and form vascular chain of lesser saphenous vein to sural nerve and vascular network of superficial and deep fascia. The superficial and deep communicating branches end at the peroneal veins.

Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.

BACKGROUND:Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cel s have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cel conditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cel s on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury.
METHODS:The bone marrow mesenchymal stem cel s were isolated from rat bone marrow. When cel s at passage 2 or 3 reached 90%confluence, the original culture medium was removed. Then the cel s were cultured in serum-free DMEM for 18 hours. After that, the culture solution was col ected as the conditioned medium of rat bone marrow mesenchymal stem cel s. Adult rats were subjected to 2 hours of right middle cerebral artery occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups:control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P>0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P<0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P<0.05), and there was also no difference between the latter two groups (P>0.05). These results suggest that rat bone marrow mesenchymal stem cel s-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.

Objective To study the mechanism of Wuzi Yanzong Wan (Five-Seed Progeny Pill) on improving heat stress resistance of sertoli cells in rats based on data from the whole genome microarray.Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan (WZYZW) group, normal group and model group.The cells in three groups were given WZYZW solution and equal volume of PBS respectively.After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 ℃ for 15 minutes, while rats in normal group were put in water bath at 37 ℃ for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technology.Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated.There were totally 473 differently expressed genes in the normal group, including 168 up-regulated genes and 305 down regulated genes.Among these down-regulated gene expressions, RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b were significant in the WZYZW group and normal group, while Dusp7 was significant among up-regulated gene expressions.The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regulation of actin cytoskeleton, MAPK signaling pathway and tight junction pathway, etc.Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of sertoli cells may involve maintenance of tight junctions between sertoli cells, inhibition of sertoli cells apoptosis and promotion of cell differentiation and maturation.

ObjectiveTo discuss the effect and mechanism of Wuzi Yanzong Wan (Five Seeds for Multiplying Pills) in the proliferation of Sertoli cells in infant rats.Methods The Sertoli cells were separated from male SD rats (20 d old) and cultured,and then randomly divided into normal group,lowdose Wuzi Yanzong Wan group (5 g/L,low-dose group),mid-dose Wuzi Yanzong Wan group (10 g/L,mid-dose group) and high-dose Wuzi Yanzong Wan group (15 g/L,high-dose group).The changes of optical density (OD) of Sertoli cells were detected by using methyl thiazolyl tetrazolium test (MTT') in all groups.The expression of Ki67 positive granules in Sertoli cells were detected by using immunohistochemistry technique in all groups.The expressions of Akt and p-Akt in Sertoli cells were detected by using Western blot method.Results Compared with normal group,the proliferation of Sertoli cells treated with Wuzi Yanzong Wan was promoted significantly (P ＜ 0.05 or P ＜ 0.01).The expression of Ki67 positive granules increased significantly (P ＜ 0.05 or P ＜ 0.01),and relative expression of p-Akt increased significantly (P ＜ 0.05 or P ＜ 0.01).Conclusion Wuzi Yanzong Wan can possibly promote the proliferation of Sertoli cells in infant rats through activating Akt signal transduction pathway and up-regulating p-Akt expression.