Transarticular bony defects after trauma and sepsis: arthrodesis using vascularized fibular transfer

Ten male patients with previously infected bony defects involving both sides of an articulation underwent arthrodesis using a vascularized fibular transfer. The average age of these patients was 38 years (range, 20 to 60 years). The size of the bony defect averaged 9 cm (range, 3 to 21 cm). The ankle was involved in five patients, the knee in two patients, the wrist in two patients, and the elbow in one patient. Nine cases represented septic pseudarthroses (eight after trauma and one after attempted ankle arthrodesis). One patient had a defect across the wrist after debridement of a chronic infection. The patients were followed for an average of 71 months (range, 26 to 144 months). Nine patients healed after the index vascularized fibular transfer, and one patient (ankle arthrodesis) required a second cancellous bone-grafting procedure for delayed union at the junction of the fibula with the talus. Four of seven patients with lower limb involvement had residual leg length discrepancies averaging 5 cm (range, 3 to 8 cm), and one had a persistent 20-degree internal rotation deformity. Two of the patients with upper limb involvement had stiff digits. Five of the nine previously employed patients returned to their former occupation (including heavy labor in four cases). Complications included two wound separations, one case of instability of the donor ankle after removal of a large fibular graft (related in part to a prior injury), and one fracture at the junction of the fibular graft with the local bone 10 months after the index procedure, which united after plate fixation and application of autogenous cancellous bone graft. Arthrodesis using a transfer of vascularized fibular bone represents a viable option for limb salvage in the face of an infected transarticular bony defect.     

Osteogenesis in cranial defects: reassessment of the concept of critical size and the expression of TGF-beta isoforms

Transforming growth factor-betas (TGF-beta) have been demontstrated to be upregulated during osteoblast function in vitro and during cranial suture fusion in vivo. The authors hypothesized that spontaneous reossification of calvarial defects was also associated with upregulation of TGF-beta. The present study was designed to (1) evaluate the concept of a critical-size defect within the calvaria in an adult guinea pig model and (2) investigate the association between the ossification of calvarial defects and TGF-beta upregulation. Paired circular parietal defects with diameters of 3 and 5 mm and single parietal defects with diameters of 8 or 12 mm were made in 45 six-month-old skeletally mature guinea pigs. Three animals per defect size were killed after survival periods of 3 days, 1 week, 4 weeks, 8 weeks, or 12 weeks. New bone ingrowth was evaluated by assessing for linear closure by a traditional linear method and by a modified cross-sectional area method using an image analysis system in which the thickness of new bone was taken into account. Immunohistochemistry was performed using rabbit polyclonal antibodies to localize TGF-beta1, -beta2, and -beta3. All specimens were photographed, and the intensity of immunostaining was graded based on subjective photographic assessment by three independent reviewers. No defect demonstrated any measurable bone replacement after a survival period of 3 days. All 3- and 5-mm defects were completely reossified after 12 weeks based on the linear analysis of new bone, indicating these defects to be less than critical size. However, new bone formation in the 5-mm defects never exceeded a mean of 40 percent by cross-sectional area of new bone. Percent of new bone formation by cross-sectional area was significantly higher within 3-mm defects than in all larger defects 4 weeks after the craniotomy, reaching a mean of 89 percent new bone by 12 weeks. Persistent gaps were noted on linear analysis of the 8- and 12-mm wounds by 12 weeks, and mean percent new bone by cross-sectional area remained below 30 percent. Immunolocalization demonstrated osteogenic fronts at the advancing bone edge and the endocranial side, in which the osteoblasts stained strongly for all isoforms of TGF-beta. The intensity of osteoblast expression waned considerably after the majority of the defect had reossified. These data indicate that histometric analysis based on cross-sectional area more accurately reflects the osteogenic potential of a cranial defect than does linear inspection of defect closure. Although the interpretation of immunolocalization studies is highly subjective, independent assessment by three reviewers indicates that isoforms of TGF-beta were upregulated during a limited "window" of time corresponding to the period of active calvarial reossification, and expression of TGF-beta corresponded to osteoblast activity within osteogenic fronts.     

Bone reconstruction of the lower extremity: complications and outcomes

A study was performed to analyze the results and final outcomes of bone reconstruction of the lower extremity. Twenty-six patients presented with type IIIB open fractures, nine with type IIIC open fractures, and 15 with chronic osteomyelitis. Seven patients underwent primary amputation, and reconstruction was attempted for 43 patients. The mean bone defect size was 7.7 cm (range, 3 to 20 cm). Bone reconstruction was achieved with conventional bone grafts in 16 cases, in association with either local (13 cases) or free (three cases) flaps. Vascularized bone transfer was performed in 24 cases, with either osteocutaneous groin flaps (10 cases), soleus-fibula flaps (12 cases), or osteocutaneous lateral arm flaps (two cases). For three patients, bone reconstruction was performed with a technique that combines the induction of a membrane around a cement spacer with the use of an autologous cancellous bone graft. Infections were observed to be responsible for prolonged hospital stays and treatment failures. The cumulative rates of sepsis were 4.6 percent at 1 week after injury and 62.8 percent at 2 months. Vascular complications were also related to infections and were responsible for four secondary amputations. One patient asked for secondary amputation because of a painful nonfunctional lower limb. Bone healing occurred in 37 of 43 cases, and the average time to union was 9.5 months, with an average of 8.7 procedures. The mean lengths of stay were 49 days for conventional bone grafts and 62 days for vascularized bone grafts. All of the 50 patients were able to walk, with an average time of 14 months. All of the patients with amputations underwent prosthetic rehabilitation. Patients mostly complained about the reconstructed limb (62.8 percent). Joint stiffness was present in 40 percent of the cases. Other long-term complications were pain (nine cases), lack of sensation (five cases), infection (five cases), and pseudarthrosis (one case). However, all of the patients with successful reconstructions preferred their salvaged leg to an amputation. Of 41 patients who were working before the injury, 26 returned to work.     

The use of anterolateral thigh perforator flaps in chronic osteomyelitis of the lower extremity

From April of 2000 to May of 2003, 28 consecutive patients with chronic osteomyelitis of the lower extremity underwent surgical debridement and reconstruction with anterolateral thigh perforator flaps (six cases were combined with vastus lateralis muscle flaps). All wounds were open for a minimum period of 6 weeks (average, 24.7 months; range, 6 weeks to 52 months). The average patient age was 42.8 years (range, 18 to 71 years), there were 21 male and seven female patients, and the average follow-up period was 18.2 months (range, 5 to 41 months). The cause of injury was an open fracture in 10 cases, secondary wound complications after reduction in eight cases, and diabetic foot in 10 cases. The surface defects ranged from 50 to 153 cm. The wounds were debrided an average of 2.5 times and then reconstructed with flap and treated with antibiotics for 6 weeks. Antibiotic beads were used in six cases and secondary bone graft procedures were performed in seven cases 3 months after the flap coverage. All 28 flaps were successful without any signs of recurrences or persistent osteomyelitis, but partial wound dehiscence was observed during early rehabilitation in two cases suspected of delayed healing caused by diabetes. These wounds healed spontaneously. All patients achieved acceptable gait function after rehabilitation. No debulking procedure was necessary in any case. Although the muscle flap is known to provide superior vascular supply, the type of flap used for coverage seems to be less critical in the final outcome, provided that total debridement and obliteration of dead spaces are achieved. A well-vascularized anterolateral thigh perforator flap was successfully used to combat infection and bring stability to wounds with chronic osteomyelitis.     

Ex vivo gene therapy in autologous critical-size craniofacial bone regeneration

In therapeutic bone repairs, autologous bone grafts, conventional or vascularized allografts, and biocompatible artificial bone substitutes all have their shortcomings. The bone formed from peptides [recombinant human bone morphogenetic proteins (BMPs)], demineralized bone powder, or a combination of both is small in size. Tissue engineering may be an alternative for cranial bone repair. In this study, the authors developed an animal model to test the hypothesis that replication-defective, adenovirus-mediated human BMP-2 gene transfer to bone marrow stromal cells enhances the autologous bone formation for repairing a critical-size craniofacial defect. The mesenchymal stromal cells of miniature swine were separated from the iliac crest aspirate and expanded in monolayer culture 1 month before implantation. The cultured mesenchymal stromal cells were infected with recombinant, replication-defective human adenovirus BMP-2, 7 days before implantation. Bilateral 2 x 5-cm2 cranial defects were created, leaving no osteogenic periosteum and dura behind. Mesenchymal stromal cells at 5 x 10(7)/ml were mixed with collagen type I to form mesenchymal stromal cell/polymer constructs. Mesenchymal stromal cells used for the control site were infected with adenovirus beta-Gal under the same conditions. After 6 weeks and 3 months, 10 miniature swine were euthanized and the cranium repair was examined. Near-complete repair of the critical-size cranial defect by tissue-engineered mesenchymal stromal cell/collagen type I construct was observed. The new bone formation area (in square centimeters) measured by three-dimensional computed tomography demonstrated that the improvement from 6 weeks to 3 months was significantly greater on the experimental side than on the control side (2.15 cm2 versus 0.54 cm2, p < 0.001) and significantly greater at 3 months than at 6 weeks (2.13 cm2 versus 0.52 cm2, p < 0.001). The difference between the experimental and control groups was significant at 3 months (mean difference, 2.13 cm2; p < 0.001). The maximal compressive strength of the new bone was similar to that of the normal cranial bone when evaluated by biomechanical testing (cranium bone versus tissue-engineered bone, 88.646 +/- 5.121 MPa versus 80.536 +/- 19.302 MPa; p = 0.227). Adenovirus was absent from all constructs by immunochemical staining at 6 weeks and 3 months after implantation. The successful repair of cranial defects in this experiment demonstrates the efficacy of the integration of the autologous stem cell concept, gene medicine, and polymers in producing tissue-engineered bone.     

Evaluating the bone regeneration in calvarial defect using osteoblasts differentiated from adipose-derived mesenchymal stem cells on three different scaffolds: an animal study

The aim of this study was to investigate the effect of three different scaffolds on the viability and differentiation of adipose-derived mesenchymal stem cells (ADMSCs) to osteoblast for bone regeneration of calvarial defect in rabbit model. Adipose was harvested from the nape of 12 rabbits by direct surgery or hollow-tip cannula. Two standardized circular calvarial defects (case and control), 8 mm in diameter each, were created in all the animals. The animals were divided into 3 different groups. In group 1 (G1), the defect was filled with polyamide + ADMSC. In group 2, poly lactic-co-glycolic acid + ADMSC was used. In group 3, decellularized amniotic membrane + ADMSC was applied. In the control defect, the non-seeded scaffolds were applied for filling the defect. Decellularized pericardial scaffolds were used as a membrane on the scaffolds. The animals were euthanized 2, 4, and 8 weeks of operation and new bone formation was assessed by different analyses. Immunohistochemical (IHC) staining with osteopontin and osteocalcin antibodies was also performed. After 2 weeks of wound healing, minimal bone regeneration was detected in all groups. Almost complete defect closure was observed in all experimental groups after 8 weeks of operation, with the greatest defect closure in the animals treated with polyamide scaffolds as compared to biopsies obtained from control defects and other experimental groups. The maximal tensile load was higher in G1, 4 and 8 weeks postoperatively, suggesting the usefulness of polyamide + ADMSC for bone regeneration in calvarial defects. Results of the IHC staining demonstrated a significant difference between seeded and non-seeded scaffold in both short- and long-term follow-ups (P < 0.05). In addition, a significant difference was observed in enhancement of IHC staining of both markers in polyamide group (seeded or non-seeded) 4 and 8 weeks postoperatively in comparison with other scaffolds. It was concluded that bone regeneration in critical calvarial defect was more successful in seeded polyamide.     

Interdental distraction osteogenesis and rapid orthodontic tooth movement: a novel approach to approximate a wide alveolar cleft or bony defect

The closure of a wide alveolar cleft and fistula in cleft patients and the reconstruction of a maxillary dentoalveolar defect in traumatic patients are challenging for both orthodontists and surgeons. This is due to the difficulty in achieving complete closure by using local attached gingiva and the great volume of bone required for the graft. In this article, the authors propose using interdental distraction osteogenesis to create a segment of new alveolar bone and attached gingiva for the complete approximation of a wide alveolar cleft/fistula and the reconstruction of a maxillary dentoalveolar defect. They performed this procedure on one patient with a traumatic maxillary dentoalveolar defect and 10 patients with unilateral or bilateral cleft lips and palates who had varied dentoalveolar clefts/fistulas. Interdental and maxillary osteotomies were performed on one side of the dental arch by the cleft or defect. After a latency period of 3 days, the osteotomized distal segment of the dental arch was then distracted and transported toward the cleft or defect by using a toothborne intraoral distraction device. The alveoli and gingivae on both ends of the cleft or defect were approximated after distraction osteogenesis. The need for extensive alveolar bone grafting was eliminated. A segment of new edentulous alveolus and attached gingiva was created interdentally at a site distant to the cleft or defect. In the cleft patients, teeth were moved orthodontically into the regenerate (newly formed alveolar bone) dental crowding 1 week after distraction. The orthodontic tooth movement was rapidly completed in 3 months, and the edentulous space was eliminated. Interdental distraction osteogenesis minimizes an alveolar cleft/fistula and helps reconstruct a maxillary dentoalveolar defect by approximating the native alveoli and gingivae; it also creates new alveolar bone and gingiva for rapid orthodontic tooth movement.     

Small intestinal submucosa in abdominal wall repair after TRAM flap harvesting in a rat model

The strength of porcine small intestinal submucosa in abdominal wall repair after transverse rectus abdominis myocutaneous flap harvesting was examined in a rat model. Changes in the levels of selected molecular markers of inflammation after small intestinal submucosa implantation were also studied. Eighty-three rats were divided into three groups. In experimental group I, an abdominal wall defect created by removal of the rectus abdominis muscle was repaired with placement of a 1.5 x 5-cm2 patch of small intestinal submucosa. In experimental group II, the muscle defect was repaired with a combination of small intestinal submucosa patch placement and fascial closure. In the control group, the defect was repaired with direct fascial closure. At postoperative times of 3 days, 2 weeks, 1 month, and 2 months, the muscle tissues adjacent to the abdominal wall repair site were subjected to biopsies for assessment of inflammation markers. Full-thickness sections of the abdominal wall from the repair site in each animal were removed for tensile strength testing and histological examinations. The results demonstrated that interleukin-6 and interferon-gamma levels were increased in the two experimental, small intestinal submucosa-treated groups at 3 days and 2 weeks postoperatively. The results of mechanical testing demonstrated that the average tensile strength of the repaired abdominal wall in the repair model with combined small intestinal submucosa placement and fascial repair was significantly greater than the values for repairs with fascial closure or small intestinal submucosa placement alone. The use of small intestinal submucosa placement in combination with fascial repair can significantly improve the strength of the repaired abdominal wall after transverse rectus abdominis myocutaneous flap harvesting.     

A case of Turner syndrome with congenital hypothyroidism untreated until age 38 years

OBJECTIVE AND METHODS: The effect of thyroid hormone on human growth and maturation is considered 'permissive'. To evaluate the effect of a prolonged thyroid hormone defect, especially in the pubertal period, a woman with untreated congenital hypothyroidism underwent studies of thyroid function and bone maturation for the first time at age 38 years 10 months and received thyroid hormone replacement. RESULTS: The karyotype was 45,X/46,XX. Menstruation had occurred for 10 years, from menarche until she was about 31 years old. Epiphyseal closure of the left hand was incomplete. The serum thyroid hormone level was virtually undetectable, and her thyroid gland was not detectable in the normal position by ultrasonography. Her height increased by 3.5 cm in the first 9 months after starting thyroid hormone replacement; after 11 months, closure of the epiphysis was complete. CONCLUSION: Thyroid hormone is necessary to achieve bone maturation and epiphyseal closure, but its role is only permissive.     

Major defect to the brachial artery following blunt trauma--a five year follow-up

A 29-year-old soldier was injured when a collapsible side of an army truck fell on his left upper arm creating lesion to the brachial artery with a defect in the artery of about 10 cm in length. Injury to one brachial vein, of the same length, was also sustained without bone or neural injury. Intraoperative Doppler ultrasonography was performed. A reverse autologous saphenous vein interposition graft with a circumference similar to that of the injured brachial artery was used to perform end-to-end anastomosis between artery and reversed vein graft. The patient returned to his military formation 16 days post operatively, with a return to full military calisthenics routine 2 months after injury. Surgical and Doppler ultrasonography control was kept regularly. Excellent late result revascularization of the injured brachial artery with reversed interposition vein graft can be seen in CT angiography five years after injury.     

One-stage reconstruction of composite bone and soft-tissue defects in traumatic lower extremities

Management of bone loss that occurs after severe trauma of open lower extremity fractures continues to challenge reconstructive surgeons. Sixty-one patients who had 62 traumatic open lower extremity fractures and combined bone and composite soft-tissue defects were treated with the following protocol: extensive debridement of necrotic tissues, eradication of infection, and vascularization of osteocutaneous tissue for one-stage bone and soft-tissue coverage reconstruction. The mechanism of injury included 49 motorcycle accidents (80.3 percent), five falls (8.2 percent), three crush injuries (4.9 percent), two pedestrian-automobile accidents (3.3 percent), and two motor vehicle accidents (3.3 percent). The bone defects were located in the tibia in 49 patients (79 percent; one patient had bilateral open tibial fractures), in the femur in seven patients (11.3 percent), in the calcaneus bone in four patients (6.5 percent), and in the metatarsal bones in two patients (3.2 percent). The size of soft-tissue defects ranged from 5 x 9 cm to 30 x 17 cm. The average length of the preoperative bony defect was 11.7 cm. The average duration from injury to one-stage reconstruction was 27.1 days, and the average number of previous extensive debridement procedures was 3.4. Fifty patients had vascularized fibula osteoseptocutaneous flaps, six had vascularized iliac osteocutaneous flaps, and five patients had seven combined vascularized rib transfers with serratus anterior muscle and/or latissimus dorsi muscle transfers. One patient received a second combined rib flap because the first combined rib flap failed. The rate of complete flap survival was 88.9 percent (56 of 63 flaps). Two combined vascularized rib transfers with serratus anterior muscle and latissimus dorsi muscle flaps were lost totally (3.2 percent) because of arterial thrombosis and deep infection, respectively. Partial skin flap losses were encountered in the five fibula osteoseptocutaneous flaps (7.9 percent). Postoperative infection for this one-stage reconstruction was 7.9 percent. Excluding the failed flap and the infected/amputated limb, the primary bony union rate after successful free vascularized bone grafting was 88.5 percent (54 of 61 transfers). The average primary union time was 6.9 months. The overall union rate was 96.7 percent (59 of 61 transfers). The average time to overall union was 8.5 months after surgery. Seven transferred vascularized bones had stress fractures, for a rate of 11.5 percent. Donor-site problems were noted in six fibular flaps, in two iliac flaps, and in one rib flap. The fibular donor-site problems were foot drop in one patient, superficial peroneal nerve palsy in one patient, contracture of the flexor hallucis longus muscle in two patients, and skin necrosis after split-thickness skin grafting in two patients. The iliac flap donor-site problems were temporary flank pain in one patient and lateral thigh numbness in the other. One rib flap transfer patient had pleural fibrosis. Transfer of the appropriate combination of vascularized bone and soft-tissue flap with a one-stage procedure provides complex lower extremity defects with successful functional results that are almost equal to the previously reported microsurgical staged procedures and conventional techniques.     

Evaluation of Freeze-Dried Dura Mater Allograft as a Collagen Based Barrier (an experimental animal study)

Freeze-dried dura mater allograft (FDDMA) was evaluated as a cost-effective collagenous barrier using an experimental animal model to determine its biocompatibility and 'cell-occlusivity' potential. The FDDMA was processed at the Civil Hospital Ahmedabad, in accordance with the guidelines provided by the South-Eastern Organ Procurement Foundation, U.S.A. Fifteen Charles Foster rats were selected for a study period of 9 weeks. The 1st set graft procedure was performed in an abdominal pouch and after 1 week bilateral mandibular defects were created. One defect was covered by the FDDMA barrier (2nd set graft procedure) and the other defect served as the control. The post operative autopsy and morphological and histological assessments were carried out at 2 and 4 weeks for the 1st set graft, and 3, 6 and 9 weeks for the 2nd set graft procedure. There was acceptance of the 2nd set graft in a pre-exposed animal and no significant inflammatory response. The processed FDDMA demonstrated optimal 'cell-occlusivity' potential for guided bone regeneration to take place in Charles Foster rats.     

Ⅳ型镶嵌式外固定器治疗下肢长骨骨缺损

目的：探讨应用IV型镶嵌式外固定器治疗下肢长骨骨缺损的临床疗效。方法：1996年4月～2008年4月应用IV型镶嵌式外固定器治疗下肢长骨骨缺损48例患者,其中股骨8例,胫骨40例。骨缺损长度为5～15cm,平均8cm。随访时间9-27个月,平均18个月。结果：48例患者肢体长度均得到恢复;骨缺损达到骨性愈合,平均愈合时间8.2个月;6例骨折成角,均〈10°;12例共25处针孔感染;所有病例无神经血管损伤表现,髋、膝、踝关节活动均未受影响。结论：IV型镶嵌式外固定器是治疗长骨复杂骨缺损、成功重建肢体长度的有效方法。     

Oromandibular reconstruction with the radial-forearm osteocutaneous flap: experience with 60 consecutive cases

One of the more difficult problems in reconstructive surgery of the head and neck is replacement of bone and soft tissue lost because of injury, osteomyelitis, or malignancy. The radial-forearm osteocutaneous flap is an accepted choice for oromandibular reconstruction. This study was undertaken to review one center's experience with 60 consecutive cases of oromandibular reconstruction with the radial-forearm osteocutaneous flap. Records of the 38 men and 22 women (mean age, 60 years; range, 26 to 86 years) were reviewed for tumor location, defect and bone length, flap failure rate, recipient- and donor-site complications, length of surgery, and hospital stay. Cancer resection was the reason for 97 percent of reconstructions; 33 percent of flaps were used to reconstruct a lateral defect of the mandible, 40 percent a lateral-central defect, and 27 percent a lateral-central-lateral defect. Mean skin flap size was 55 cm2 (range, 15 to 117 cm2) and mean bone length, 9.4 cm (range, 5 to 14 cm). The microvascular success rate was 98.3 percent. Complications included fracture of the donor radius (15 percent), nonunion of the mandible (5 percent), and hematoma (8.3 percent). These results are comparable to results reported in the literature with other radial forearm flaps. The free radial osteocutaneous flap is a safe and reliable choice for mandibular reconstruction. It offers sufficient bone to reconstruct large defects and can provide adequate pedicle length for vessel anastomosis to the contralateral side of the neck. The above attributes make the radial forearm osteocutaneous flap one of the "first line" flap choices for oromandibular reconstruction.     

Bone defect repair in rat tibia by TGF-β1 and IGF-1 released from hydrogel scaffold

Bone repair is one of the major challenges facing reconstructive surgery. Bone regeneration is needed for the repair of large defects and fractures. The ability of TGF-β1 and IGF-1 incorporated into hydrogel scaffold to induce bone regeneration was evaluated in a rat tibia segmental defect model. External fixation was performed prior to the induction of the segmental bone defect in order to stabilize the defect site. Hydrogel scaffold containing either TGF-β, IGF-1, TGF-β + IGF-1, hydrogel containing saline or saline, were inserted in the defect. Calcified material was observed in the defects treated with TGF-β 2 weeks following the start of treatment. Bone defects treated with TGF-β, IGF-1 or TGF-β + IGF-1 revealed significant bone formation after 4 and 6 weeks when compared to the control specimens. X-ray images showed that solid bone was present at the defect site after 6 weeks of treatment with TGF-β or TGF-β + IGF-1. A less pronounced bone induction was observed in the control specimens and bones treated with IGF-1. Percent closure ratio of bone defects after 6 weeks were 40, 80, 89, and 97% for saline, hydrogel, IGF-1, TGF-β and IGF-1 + TGF-β groups, respectively. It is concluded that hydrogel scaffold can serve as a good osteoconductive matrix for growth factors, and that it provides a site for bone regeneration and enhances bone defect healing and could be used as alternative graft material. This revised version was published online in July 2006 with corrections to the Cover Date.     

Observation of reproductive cycle of female yellow-margined box turtle (Cuora flavomarginata) using radiography and ultrasonography

This study presents a combination of radiography and ultrasonography to observe the reproductive cycle of 24 captive female yellow-margined box turtles in Taiwan. Radiography was mainly used to monitor clutch size, whereas ultrasonography was applied to detect changes in the follicles throughout the year. The observation of the 24 female turtles was performed from April 2007 to June 2008. Their average carapace length was 16.62 ± 1.66 cm and their average body weight was 812 ± 164.98 g. The mean clutch size was three (87 eggs/29 clutches) and the reproductive frequency was 95.83% (23/24). Double clutches were detected in 79.2%, and 20.8% had single clutches. Ovulation occurred from March through August, and the average follicular diameter was 2.16 ± 0.18 cm. Follicles entered the latent period in October (at 1.54 ± 0.26 cm), and vitellogenesis of the next reproductive cycle began in November. Using radiography, the eggshell could be detected on the ninth day after ovulation. The average period of the single clutch group was 6.9 weeks (range 5.1-8.5 weeks). In the double clutch group, the average period of the first clutch was 5.5 weeks (range 4-7.8 weeks) and that of the second clutch was 5.2 weeks (range 4-7.8 weeks). This study has advanced the understanding of reproductive physiology of yellow-margined box turtle and established a valuable and practical model for comparative study of the reproductive physiology of other chelonians.     

自固化磷酸钙人工骨修复小儿局部骨缺损的临床应用

目的：研讨自固化磷酸钙人工骨（Calcium Phosphate Cement,CPC)填充修复小儿局部骨缺损的临床意义,方法：选用CPC修复小儿骨缺损18例,年龄最小8个月,最大12岁,平均8岁,骨缺损部位：肱骨9例,胫骨6例,胫骨3例,病因,单纯性骨囊肿8例,骨纤维异常增生症5例,动脉瘤样骨囊肿4例,嗜酸性肉芽肿1例,骨缺损大小,最大7cm,最小2cm,平均5cm,CPC填充方式：单纯粉末7例,粉末＋松质骨粒6例,粉末＋条形骨块5例。CPC初步固化时间,最短15分钟,最长30分钟,平均20分钟,随访时间：13－27个月,平均18．5个月。结果：全组18例应用CPC后未见明显局部和全身不良反应。手术前后血PH值钙磷代谢无异常改变。X线片显示：CPC与宿主骨接触紧密,无脱落,术后3个月出现降解,新生骨形成。结论：CPC安全无毒,使用方便,易塑形,生物相容性好,能在体内降解,可以替代自体骨材料在小儿局部骨缺损应用。     

Activity,movements, and sociality of newborn Mongolian gazelle calves in the Eastern Steppe

We hand-captured, fitted with motion-sensitive VHF transmitters, and monitored 19 newborn (1–2 days old) Mongolian gazelle Procapra gutturosa Pallas, 1777 calves in Dornod, Mongolia during the 2000 and 2003 calving seasons to identify changes in activity, movements, and sociality with age. Overall, activity was highly variable throughout the day, regardless of age or year. Calf activity increased, however, from day 2 (age = 2 days; 18% total activity) to day 7 (29%), and 3-fold by day 25 (54%). By days 5–8, calves had moved an average of 6.6 km (range: 2–21 km) from capture sites but were still only seen alone or with their mother. By day 24–26, however, they were located an average of 41 km (range: 24–63 km) away from their capture site, and most (7 of 9) were in aggregations of >1000 animals. Mongolian gazelles do not display behaviour associated with “hiders” after about 2–3 weeks. This reflects the wide-ranging, nomadic existence of the species.     

骨髓干细胞动员对动脉粥样硬化破裂斑块的稳定与修复作用

目的研究自体骨髓干细胞动员对兔动脉粥样硬化（AS）破裂斑块的稳定与修复作用。方法用液氮冻伤术创建兔AS破裂斑块模型,动员组注射重组人粒细胞刺激因子（rhG-CSF）动员自体骨髓干细胞,对照组注射等量生理盐水,连续5 d。动员第5天抽血分离获取单个核细胞,BrdU标记后经静脉注入动物体内;分别于动员后3d和4周末抽血,ELISA法检测兔血清MMP-9、hsC-RP及PAI-1水平;动员后4周处死兔,HE染色和Masson三色染色观察斑块病理形态,免疫组化染色观察BrdU在斑块区表达情况。结果动员5 d后,动员组兔外周血有核细胞计数及单核细胞比例明显增高;动员后4周,动员组新生内皮细胞及胶原纤维明显增多,在斑块区发现有BrdU标记的阳性细胞,动员组血清MMP-9、hsC-RP及PAI-1水平明显降低。结论应用rhG-CSF动员自体骨髓干细胞能通过促进血管内皮细胞和胶原纤维再生,降低炎症因子及凝血纤溶因子而稳定与修复AS破裂斑块。     

Programmed cell death in post-traumatic bone callus.

Some osteoblasts in the expanded population of periosteal cells that occurs following bone injury are removed from the callus by apoptosis. Our objective was to study whether the consequences of activation of the death program could include feedback control of the healing response. Transforming growth factor beta and interleukin-1beta were delivered together continuously to a standardized tibial defect in rats for 3 days using implanted micro-osmotic pumps. The bones were recovered at 1, 2, 3, 5, 7, 10 and 14 days after injury (n = 6 in each treated and control group) and concentrations of proliferating cells, osteoblasts and apoptotic bodies were determined. The injury-induced apoptotic component of the healing response was shifted in time due to the combined cytokines, compared with vehicle only, with the result that the peak in the concentration of apoptotic bodies occurred 2-3 days earlier in the treated animals. Neither osteoprogenitor proliferation nor osteoblast concentration was affected by addition of the cytokines. The results suggested that activation of apoptosis during injury repair was not necessarily a passive consequence of the cellular response to injury. Programmed cell death could therefore have an active role in modulating bone repair.