Objective The clinical effect of the lateral femoral cutaneous artery flap for soft tissue defects of leg wounds. Methods From October 2007 to January 2016, VSD was firstly used to promote the growth of granulation tissue. When the growth of granulation tissue became satisfactory, flaps were designed based on the anatomical characteristics of the lateral femoral cutaneous artery. We repaired 20 cases of wound defects by cutting flaps that coincide with the recipient vessels. Result 20 cases were followed up for 6 to 24 months, 12 months on average. All flaps were survived and only one case had small area of necrosis flap which was healed by replacing medicines. In all cases, wounds were healed and flaps showed good color and good texture. The strength of quadriceps muscle was good and the extension of knee flexion was 0° to 150°. Conclusion To The lateral femoral cutaneous artery flap is used for soft tissue defects of refractory wounds on leg , flap donor sites are sutured directly, the treatment period is shorten and the method is safe and effective. The lateral femoral cutaneous artery flap is one of ideal choices for wound tissue defects.

Objective To master the prevalence of plague and its trend in Guizhou Province,and to analyze the plague monitoring results from 2000 to 2012.Methods The report of infectious disease,the information of plague natural focus and the epizootic monitoring data of Xingyi City,Anlong County and Dingxiao Distract of Guizhou Province from 2000 to 2012 were collected and the status of the plague natural focus was analyzed.Results There were 137 cases of gland plague in Xingyi City and Arlong county from 2000-2003,1 death,and mt plague occurred in 66 villages.Fifty-four strains of Yersiniapestis were detected and 49 rats were plague antigen F1 positive(49/160).No human plague occurred between 2004-2012.A total of 4 plague antigen F1 positive rats were detected in Dingxiao District and Xingyi City in 2005 and 2006.There was no Yersinia pestis and F1 antibody in 2007-2012.The epidemic stage of plague was from 2000-2003; the active stage was from 2004-2006; and the quiescent stage was from 2007-2012.The dominant species of the plague natural focus was Rattus flavipestus (42.83％,7 966/18 597),but was replaced by Rattus norvegicus at the epidemic stage (47.22％,1 480/3 134) and the active stage(35.35％,2 071/5 196).The density of rodents was 5.34％ at the epidemic stage,which was higher than that of the active stage (3.27％) and the quiescent stage (1.71％,x2 =2 286.15,P ＜ 0.01).Xenopsylla cheopis(56.34％,10 034/17 811) was the dominant species,and the index was 1.537 9,which was greater than those of the active stage(0.959 6) and the quiescent stage(0.540 4,x2 =492.68,P ＜ 0.01).Conclusions The plague of Guizhou Province is at the quiescent stage.Both the density of rodents and the Xenopsylla cheopis index are lower than the national standard of controlling.

Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; mortality ; Humans ; Infant ; Male

BACKGROUND:Open fracture of lower limb with severe soft tissue and bone defects also accompanies anterior tibial soft tissue defects and exposure of sclerotin and steel plate, which can be crucial y treated with strong fixation and use of skin flap to block the wound.
OBJECTIVE:To explore the clinical efficacy of a large area of soft tissue defects in the anterior tibia using vacuum sealing drainage combined with groin free flap.
METHODS:A total of 24 patients with a large area of soft tissue defects in the anterior tibia were included in this study and then divided into two groups, with 12 cases in each group. In vacuum sealing drainage group, the scope of soft tissue defects was ranged from 10 cm×15 cm to 15 cm×20 cm. After the debridement, the fracture was fixed with external fixation scaffold and the wound was covered with the vacuum sealing drainage dressing. The blood clot was rinsed with normal saline via T-tube, and 7-10 days later the vacuum sealing drainage was given. According to the growth of granulation tissue, the wound was secondarily sutured, fol owed by groin free skin flap of superficial iliac circumflex artery with medial knee arteriovenous anastomosis transplantation. In the non-vacuum sealing drainage group, the wound size was ranged from 10 cm×5 cm to 30 cm×20 cm, the period from injury to admission was 1-24 hours. They were given conventional debridement and secondary fixation or skin flap transplantation.
RESULTS AND CONCLUSION:The length of preoperative hospital stay and the skin flap are in vacuum sealing drainage group were significantly better than those in non-vacuum sealing drainage group (P<0.05). There was no significant difference in the length of postoperative stay and total length of hospital stay between the two groups (P>0.05). The wound infection rate was 0 in vacuum sealing drainage group and 75%in non-vacuum sealing drainage group at 8-14 days after treatment. The wound and donor area incision were healed at I stage, the skin grafts survived. Al the involved patients in two groups were fol owed up, for 6-36 months. During the fol ow-up process, the fracture healing time in non-vacuum sealing drainage group was significantly longer than that in vacuum sealing drainage group. The skin flap in two groups was similar to surrounding skin in color and texture, the flap exhibited no vessels, no ulceration, and no clumsy. The vacuum sealing drainage combined with groin free flap can timely control a large area of soft tissue defects post-trauma, improve wound blood supply, shorten preoperative preparation time, early close the wound, significantly promote the healing of wound and fracture. The skin flap is soft, flexible, wel-looking, and active functional, it significantly shortens the course of treatment and maximizes the recovery of limb function.

Objective To observe the effects of transplantation of autologous adipose-derived stem cells (ASCs) on osteoporosis (OP) in a rabbit ovariectomy (OVX) model.Methods A total of fifteen 6-month-old female New Zealand white rabbits were randomly divided into two groups:ovariectomy group (group A,n =12) and sham operation group (group B,n =3).All rabbits were subjected to bilateral ovariectomy in the group A.Six months later,bone mineral density (BMD) of group A and group B were measured by dual energy X-ray absorptiometry (DXA) to check the result of OVX-OP.ASCs harvested from adipose of OP rabbits were cultured to be expanded and differentiated in osteogenic medium in vitro.Osteogenesis was evaluated by alizarin red staining,alkaline phosphatase (ALP) staining and quantitative assays of osteocalcin (OCN).Autologous osteo-induced ASCs were mixed in calcium alginate hydrogel (CAH) and then transplanted in the left distal femurs,while CAH was transplanted in the right distal femurs of OP rabbits.At 12 weeks after implantation,BMD,micro-CT and histomorphological analyses were performed on these rabbits.Results The BMD of femurs in group A rabbits were obviously lower than that of group B rabbits (P ＜ 0.05) at 6 months after OVX.Compared with control group,ASCs cultured in osteo-induction medium had similar proliferation rate as the non-induced cells,but displayed positive ALP and alizarin red staining and OCN contents.At 12 weeks after implantation,the cell-treated femurs displayed higher BMD,bone trabecula number,trabecula thickness and separation than those of control group,while the structure model index and porosity were lower (P ＜ 0.05).Histological examination indicated that the trabecular thickness increased with complete CAH resorption in cell-treated group,while CAH remained in control group.Conclusions Transplantation of autologous ASCs can help strengthen osteoporotic bone in OVX-OP rabbits,providing a novel approach to OP treatment.

Objective To analyze the effects of Leptospira interrogans ( L. interrogans) infection on the activation of NLRP3 in THP-1 and J774A. 1 cells and to further understand the mechanism of inflam-mation caused by L. interrogans in different hosts. Methods Human mononuclear macrophage cell line (THP-1) and murine mononuclear macrophage cell line (J774A. 1) were infected with L. interrogans strain 56601. The expression of NLRP3 at mRNA and protein levels were measured by using real-time RT-PCR and flow cytometry analysis, respectively. The NLRP3-mediated secretion of IL-1β, IL-18 and IL-33 was detec-ted by ELISA combined with the NLRP3 inhibitory test. Results Compared with the normal cells, the ex-pression of NLRP3 at mRNA level in L. interrogans-infected THP-1 cells was respectively increased by 4. 05, 0. 34, 0. 33, 0. 06 and 1. 66 times at the time points of 1 h, 2 h, 4 h, 12 h and 24 h after infection ( P<0. 05), while that in L. interrogans-infected J774A. 1 cells was respectively increased by 12. 98, 16. 19, 10. 68, 5. 8 and 0. 57 times (P<0. 05). The expression rates of NLRP3 protein in THP-1 and J774A. 1 cells respectively increased from 9. 26% to 94. 01%, 89. 24%, 31. 80%, 19. 74%, 11. 28% and from 18. 71%to 58. 78%, 43. 64%, 36. 42%, 76. 46%, 85. 21% at the time points of 1 h, 2 h, 4 h, 12 h and 24 h af-ter L. interrogans infection (P<0. 05). The level of IL-1β in L. interrogans-infected THP-1 cells was 73. 07 pg/ml, 939. 24 pg/ml, 939. 24 pg/ml, 843. 22 pg/ml and 851. 06 pg/ml at the time points of 1 h, 2 h, 4 h, 12 h and 24 h, respectively (P<0. 05), while the level of IL-1β in L. interrogans-infected J774A. 1 cells began to rise at the time point of 12 h from 191. 17 pg/ml to 254. 4 pg/mL at the time point of 24 h (P<0. 05). The level of IL-18 in L. interrogans-infected THP-1 cells was 913. 89 pg/ml, 808. 19 pg/ml, 483. 54 pg/ml, 204. 19 pg/ml and 189. 09 pg/ml at the time points of 1 h, 2 h, 4 h, 12 h and 24 h, re-spectively (P<0. 05), while the level of IL-18 in L. interrogans-infected J774A. 1 cells increased at the time point of 24 h, which was 113. 37 pg/ml (P<0. 05). A slight increase in the level of IL-33 was detected in L. interrogans-infected J774A. 1 cells at the time points of 12 h and 24 h to 201. 14 pg/ml and 155. 68 pg/ml, respectively (P<0. 05), but no significant change was detected in L. interrogans-infected THP-1 cells (P>0. 05). Results of the inhibitory test showed that the up-regulation of IL-1β , IL-18 and IL-33 in THP-1 and J774A. 1 cells were effectively inhibited by the specific inhibitor of NLRP3. Conclusion NL-RP3 inflammasome was activated and involved in the production of specific inflammatory cytokines IL-1βand IL-18 in both THP-1 and J774A. 1 cells after L. interrogans infection, but the inflammatory cytokines induced by L. interrogans infection varied in different cells. L. interrogans induced earlier and higher level of IL-1βand IL-18 production in human macrophages than in murine macrophages.

Objective To investigate the expression of inducible nitric oxide synthase ( iNOS) and the levels of nitric oxide (NO) in THP-1 and J774A. 1 cells during Leptospira interrogans (L. interrogans) infection for a better understanding of the mechanism of macrophages involved defense against L. interrogans strains in different hosts. Methods The human mononuclear macrophages (THP-1) and the murine mono-nuclear macrophages (J774A. 1) were infected with L. interrogans strain 56601. The expression of iNOS at mRNA and protein levels were determined by using real-time RT-PCR and flow cytometry analysis. The lev-els of NO were detected with Griess test. Results The expression of iNOS at mRNA level in J774A. 1 and THP-1 cells infected with L. interrogans strains for 2, 4, 12 and 24 hours were respectively 1. 37, 2. 82, 25. 76, 27. 47 times and 1. 59, 3. 98, 3. 89, 8. 81 times than that of cells without infection (P<0. 05). The expression rates of iNOS protein in J774A. 1 cells were increased from 34. 16% to 85. 85%, 93. 82%, 91. 77% and 93. 65% along with the increased time of infection time (P<0. 05). The expression rates of iNOS protein in THP-1 cells were up-regulated from 22. 08% to 72. 64%, 81. 33%, 80. 03% and 65. 72%after 2, 4, 12 and 24 hours of infection (P<0. 05), respectively. Results of the Griess test indicated that the levels of NO in J774A. 1 and THP-1 cells were respectively increased from 0. 1588 μmol/L to 0. 2208μmol/L, 0. 2668μmol/L, 0. 3808μmol/L, 0. 3828μmol/L and from 0. 0988μmol/L to 0. 2848μmol/L,0. 3228 μmol/L, 0. 2608μmol/L and 0. 3308μmol/L after infection with L. interrogans strains for 2, 4, 12 and 24 hours (P<0. 05). Conclusion The expression of iNOS and the levels of NO in J774A. 1 and THP-1 cells were significantly increased during L. interrogans infection. This study might help to explain the bactericidal mechanism of macrophages derived from different hosts against L. interrogans infection.

Objective To investigate the effects of Leptospira interrogans ( L. interrogans) infec-tion on the activities of NADPH oxidase ( nicotinamide adenine dinucleotide phosphate-oxidase) and the lev-els of reactive oxygen species (ROS) in THP-1 and J774A. 1 cells and to understand the bactericidal mecha-nisms of macrophages in different hosts against L. interrogans. Methods Human mononuclear macrophage cell line (THP-1 cells) and murine mononuclear macrophage cell line (J774A. 1 cells) were infected with L. interrogans strain 56601. The activities of NADPH oxidase and the levels of superoxide ion ( O-2 ) were measured with spectrophotography. Changes in the levels of ROS were detected with immunofluorescence as-say. Results Compared with the normal cells, the activities of NADPH oxidase in L. interrogans-infected J774A. 1 cells changed from 0. 619 0 μmol · min-1 · mg-1 to 0. 305 5 μmol · min-1 · mg-1 , 6. 141 5μmol·min-1 ·mg-1 , 1. 487 1μmol·min-1 ·mg-1 and 0. 964 6μmol·min-1 ·mg-1 after 2, 4, 12 and 24 hours of infection, respectively (P<0. 05), while the activities of NADPH oxidase in L. interrogans-infected THP-1 cells were up-regulated from 0. 723 5μmol·min-1 ·mg-1 to 0. 884 2μmol·min-1 ·mg-1 , 1. 897 1μmol·min-1 ·mg-1 , 1. 125 4 μmol·min-1 ·mg-1 and 0. 562 7 μmol·min-1 ·mg-1 , respectively ( P<0. 05). The levels of O-2 in L. interrogans-infected J774A. 1 cells at the time points of 2 h, 4 h, 12 h and 24 h after infection increased from 0. 189 0μmol/L to 0. 236 3μmol/L, 0. 297 7μmol/L, 0. 324 0μmol/L and 0. 305 7 μmol/L, respectively (P<0. 05), while the levels of O-2 in L. interrogans-infected THP-1 cells rose from 0. 123 7 μmol/L to 0. 149 3 μmol/ L, 0. 249 0 μmol/ L, 0. 270 0 μmol/ L and 0. 272 7μmol/L, respectively (P<0. 05). The fluorescence intensity of ROS in THP-1 and J774A. 1 cells increased gradually after infection with L. interrogans for 2 h and decreased after reaching the peak at 24 h. Conclu-sion Both the activities of NADPH oxidase and the levels of O-2 in J774A. 1 and THP-1 cells were signifi-cantly upregulated after infected with L. interrogans, especially in J774A. 1 cells. The results of this study provided references for further elucidating the bactericidal mechanisms of macrophages in different hosts against L. interrogans.

BACKGROUND: Cell-based therapies for treating bone defects require a source of stem cells with osteogenic potential. There is evidence from pathologic ossification within muscles that human skeletal muscles contain osteogenic progenitor cells. However, muscle samples are usually acquired through a traumatic biopsy procedure which causes pain and morbidity to the donor. Herein, we identified a new alternative source of skeletal muscle stem cells (SMSCs) without conferring morbidity to donors. METHODS: Adherent cells isolated from human orbicularis oculi muscle (OOM) fragments, which are currently discarded during ophthalmic cosmetic surgeries, were obtained using a two-step plating method. The cell growth kinetics, immunophenotype and capabilities of in vitro multilineage differentiation were evaluated respectively. Moreover, the osteogenically-induced cells were transduced with GFP gene, loaded onto the porous β-tricalcium phosphate (β-TCP) bioceramics, and transplanted into the subcutaneous site of athymic mice. Ectopic bone formation was assessed and the cell fate in vivo was detected. RESULTS: OOM-derived cells were fibroblastic in shape, clonogenic in growth, and displayed phenotypic and behavioral characteristics similar to SMSCs. In particular, these cells could be induced into osteoblasts in vitro evidenced by the extracellular matrix calcification and enhanced alkaline phosphatase (ALP) activity and osteocalcin (OCN) production. New bone formation was found in the cell-loaded bioceramics 6 weeks after implantation. By using the GFP-labeling technique, these muscle cells were detected to participate in the process of ectopic osteogenesis in vivo. CONCLUSION: Our data suggest that human OOM tissue is a valuable and noninvasive resource for osteoprogenitor cells to be used in bone repair and regeneration.
Alkaline Phosphatase ; Animals ; Biopsy ; Extracellular Matrix ; Fibroblasts ; Humans* ; In Vitro Techniques* ; Kinetics ; Methods ; Mice ; Mice, Nude ; Muscle Cells ; Muscle, Skeletal ; Muscles ; Ossification, Heterotopic ; Osteoblasts ; Osteocalcin ; Osteogenesis* ; Regeneration ; Stem Cells* ; Tissue Donors

Objective To construct a prediction model for the risk of ischemic stroke (IS) by classification tree model, and evaluate its application value. Methods By cluster sampling, 858 IS patients with perfect clinical data from January to December 2017 in the Affiliated Hospital of Guilin Medical College (IS group) were enrolled, and 844 health checkups matched with the gender and age of IS patients in the same period were enrolled as controls (healthy control group). The metabolic characteristics of the two groups were compared and analyzed. The classification tree model was used to construct the prediction model of the risk of IS, and the gain diagram, index chart, risk value of misclassification probability and receiver operating characteristic curve (ROC) were used to evaluate the application value of the model. Results Compared with the healthy control group, body mass index (BMI), fasting blood glucose (FPG), triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) in IS group were significantly increased [BMI (kg/m2): 25.34±3.70 vs. 24.24±3.10, FPG (mmol/L): 6.79±2.89 vs. 5.73±1.17, TG (mmol/L):1.62±1.06 vs. 1.44±1.06, TC (mmol/L): 4.70±2.73 vs. 4.35±0.79, LDL-C (mmol/L): 3.18±0.94 vs. 2.73±0.73, all P < 0.01], high density lipoprotein cholesterol (HDL-C) was significantly decreased (mmol/L: 1.12±0.33 vs. 1.35±0.36, P < 0.01), and the proportion of hypertension, smoking and drinking were significantly increased (69.0% vs. 41.9%, 23.1% vs. 16.8%, 19.2% vs. 13.4%, all P < 0.01). By assigning values to each factor [IS: No = 0,Yes = 1; BMI: < 24.0 kg/m2=0, ≥ 24.0 kg/m2= 1; FPG : < 7.0 mmol/L = 0, ≥7.0 mmol/L = 1; TG: < 2.26 mmol/L = 0, ≥2.26 mmol/L = 1; TC: <6.22 mmol/L = 0, ≥6.22 mmol/L = 1; LDL-C: < 4.14 mmol/L = 0, ≥4.14 mmol/L = 1; HDL-C: < 1.04 mmol/L = 0, ≥1.04 mmol/L = 1; hypertension: No = 0,Yes = 1; smoking: No = 0,Yes = 1; drinking: No = 0,Yes = 1], a classification tree model was established to analyze the risk factors of IS. The classification tree model consisted of 4 layers and 17 nodes: the first layer was hypertension, the second layer was FPG and HDL-C, the third layer was HDL-C and FPG, and the fourth layer was LDL-C and smoking. There were five explanatory variables screened out in the model, including hypertension, FPG, HDL-C, LDL-C and smoking. The first layer of the tree showed that the incidence of IS in hypertensive population (62.6%) was significantly higher than that in non-hypertensive population (35.2%). The second layer of the tree showed that the incidence of IS in people with hypertension with HDL-C≥1.04 mmol/L (53.6%) was lower than that in people with HDL-C < 1.04 mmol/L (78.5%). However, in the population without hypertension, the probability of IS occurrence in the population with FPG ≥ 7.0 mmol/L (71.1%) was significantly higher than that in the population with FPG < 7.0 mmol/L (28.3%). The third layer of the tree showed that the IS incidence of HDL-C ≥1.04 mmol/L (21.8%) was lower than that of HDL-C < 1.04 mmol/L (48.7%) in the population without hypertension and FPG < 7.0 mmol/L. However, in the population with hypertension and HDL-C ≥ 1.04 mmol/L, the probability of IS occurrence in the population with FPG ≥ 7.0 mmol/L (78.6%) was significantly higher than that in the population with FPG < 7.0 mmol/L (46.7%). The fourth layer of the tree showed that the IS incidence of people with LDL-C ≥4.14 mmol/L (53.8%) was higher than that of people with LDL-C < 4.14 mmol/L (19.0%) in the population without hypertension, FPG < 7.0 mmol/L and HDL-C ≥ 1.04 mmol/L. In the population without hypertension, the incidence of IS in smokers (76.9%) was higher than that in non-smokers (39.1%) of people with FPG < 7.0 mmol/L and HDL-C <1.04 mmol/L. In the population with hypertension, the probability of IS occurrence in the population with LDL-C ≥4.14 mmol/L (72.5%) was higher than that in the population with LDL-C < 4.14 mmol/L (44.4 %) of people with HDL-C ≥ 1.04 mmol/L and FPG < 7.0 mmol/L. The gain diagram of IS classification tree model shown that the gain value increased rapidly from 0% to 100% and then tended to be stable. The index chart shown that the index value kept stable in the moving direction from above 100% and then dropped rapidly to 100%, indicating the model was very well. The risk value of misclassification probability of the classification tree model was 0.291, and the correct rate of risk factor for IS patients was 70.90%. The area under ROC curve (AUC) was 78.0% [95% confidence interval (95%CI) =75.9%-79.9%, P < 0.001], the sensitivity was 62.5% (95%CI = 59.1%-65.7%) and the specificity was 79.4% (95%CI =76.5%-82.1%). Conclusion Classification tree model can properly predict the risk factor of IS, and the most important risk factors are hypertension, hyperglycemia, high LDL-C and smoking.