BACKGROUND: Chitin and its derivatives possess many biological activities such as immune modulation and promoting wound healing and hemostasis. As one of the derivatives of chitin, the effect of N-acetylglu cosamine on fracture healing has not yet been acknowledged.OBJECTIVE: To observe the changes in osteoblastic activity and the expression of bone morphogenetic protein (BMP) in response to the intervention with N-acetylglucosamine during the fracture healing. DESIGN: A randomized controlled experiment SETTING: Department of Anatomy, Medical College of Qingdao University.MATERIALS: This experiment was carried out in the Laboratory of Neuroanatomy, Medical Colloge of Qingdao University from March to August 2002. Seventy healthy New Zealand rabbits were randomized into four groups, namely saline group (n=23), Jiegupian group (n=23) and N-acetyl glucosamine group (n=24).METHODS: The rabbits were subjected to operations to induce 3-mm bone defect in the middle segment of the right radius with subsequent treatments with saline (1.00 mL/kg), Jiegupian (1.00 mL/kg) and N-acetyl glucosamine (0.28 g/kg) via intragastric administration on a daily basis till the time of tissue sampling at postoperative days 9, 17, 30 and 42, respectively. X-ray examination and HE staining were performed to observe the healing process, and the expression of BMP was assessed immunohistochemically.MAIN OUTCOME MEASURES :① General condition of the rabbits after operation; ② Fracture healing process revealed by X-ray at the specified time points; ③ Expression of BMP between the fracture ends at each time point; ④ Results of histological observation.RESULTS: According to intent-to-treat analysis, 70 rabbits entered the data analysis. X-ray revealed better fracture healing in N-acetylglucosamine group than in the ither two groups at postoperative days 9, 17, 30 and 42(P ＜ 0.05). Compared with saline group, the expression of BMP protein in Jiegupian group and N-acetylglucosamine group was increasing at postoperative 9 and 17 days, particularly in the latter group (P ＜ 0.05), but decreased at postoperative 30 and 42 days. Histologically, the gap between the fracture ends reduced in N-acetylglucosamine group at postoperative 9 days with the appearance of ()bmogenous bone trabecula an orderly arrangement of osteoblasts; in saline group, the gap was greater and contained less or even no bone trabecula, with much less, if any, osteoblasts.On postoperative day 30, the bone defect were completely filled by bone trabecula in N-acetylglucosamine group, with the bone trabecula almost communicating with the bone marrow cavity, and the osteoblasts surrounding the bone trabecula were arranged orderly with osteoclasts seldom observed; in saline group, the defect was substituted mostly by bone trabecula with breaches, and the chondrocytes could be observed scarcely. On postoperative day 42, the bone plate woven bone formed primarily at the fracture ends in N-acetylglucosamine group, with completely communicating marrow cavity and numerous active osteoclasts, and osseous lacuna can be observed; in saline group, the defect was filled with bone trabecula but clear fusion lines were not seen, and communication of the bone marrow cavity was nearly achieved with orderly arranged osteoblasts and presence of the osteoclasts.CONCLUSION: N-acetylglucosamine can accelerate fracture healing possibly by promoting the osteoblastic activity and increase the expression of BMP during the early period of fracture healing.

BACKGROUND: Both apoptosis suppression gene Bcl-2 and apoptosis in duction gene Bax take parts in the apoptosis of neurons in ischemic penum bra. Whether would the polypeptide from chlamys farreri that is proved to be of anti-oxidation and anti-apoptosis in vitro protect the ischemic neurons from apoptosis? OBJECTIVE: To observe the effect of chlamys farreri on the Bcl-2 and Bax protein-associated apoptosis in penumbra and its role in neuron protection. DESIGN: A randomized trial.SETTING: Department of Anatomy, Medical College of Qingdao University.MATERIALS: The trial was conducted in Nerve Anatomy Laboratory of Medical College of Qingdao University from March to April 2000. The subjects were 32 adult Wistar rats that were randomly and averagely assigned into 4 groups: polypeptide chlamys farreri group, sterile water group, model control group and sham group. The chlamys farreri was provided by the Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences.INTERVENTIONS: Model of brain ischemia and reperfusion was made in rats in polypeptide chlamys farreri, sterile water and model control group by occlusion of middle cerebral artery. The model was not established in rats in sham group. The rats in chlamys farreri group received intraperitoneal injection of chlamys farreri of volume fraction 0. 1 at the dose of 0. 1 mL/kg each day for 2 days prior to modeling and an extra injection 15 minutes just before modeling. And the rats in sterile water group received intraperitoneal injec tion of sterile water with the dose of 0. 1 mL/kg each for two days and an extra injection 15 minutes before modeling. Those in model control group and sham group were exposed to nothing. Then models were established in rats in chlamys farreri, sterile water and model control group by inserting 4-0 nylons sutures from external carotid artery through bifurcation of carotid artery,extracranial and intracranial segments of internal carotid artery till the initial part of middle cerebral artery to make acute ischemia of middle cerebral artery perfusion area. The model was considered successful by the presentation of Horner' s syndrome, adduction-flexion of right forearm when tail being lifted and right turning during walk of the rat. The rats in sham group underwent the same procedures as that in the other groups except for the occlusion of middle cerebral artery. Then brains of the rats were taken for immunohistochemical determination of Bcl-2 and Bax proteins. The protein expression was expressed by absorbance of the products of their immunological reaction.MAIN OUTCOME MEASURES: The expression differences betweenBcl-2 and Bax proteins in penumbras of the groups.RESULTS: There were 32 rats entered the stage of analysis after complement of subjects. ① Bcl-2 expression in penumbra: The absorbance in model control group and sterile water group were higher than that in sham group (0.453±0.048,0.510±0.061,0.211±0.023, F=683.78, q=21.13 to 24.74, P ＜ 0.01), and that in chlamys farreri group(0. 954 ±0. 059) was more than that in model control group and sterile water group( q = 38.08 to 41.69, P ＜ 0.01) . ② Bax expression in penumbra: The absorbance in model control group and sterile water group were higher than that in sham group (0. 834 ±0. 082, 0. 790 ±0. 102, 0. 125 ±0. 017, F=590.44, q =49.57 to 51.98, P ＜ 0.01 ) ] and that in chlamys farreri group (0.471 ± 0. 045 ) was suppressed as compared with that in model control group and sterile water group(q=23. 80 to 26. 23, P ＜0. 01).CONCLUSION: Chlamys farreri is capable of increasing Bcl-2 protein and decreasing Bax protein in cerebral penumbra to brake the initiation of neuron apoptosis after ischemia-reperfusion and preserve neuronic function in penumbra.

BACKGROUND: Cryotherapy of acute soft tissue injury has been widely used in clinical practice.OBJECTIVE: To observe the histological changes and treatment effect of different cryotherapies on the rats' acute damage of soft tissue. METHODS: Neonatal Wistar rats were randomized to normal, model, intermittent cryotherapy and continuous cryotherapy groups. Models of acute damage of soft tissue were established in model, intermittent cryotherapy and continuous cryotherapy groups. In intermittent cryotherapy group, the injury was treated by intermittent cryotherapy with ice bag at 4 °C; in the continuous cryotherapy group, the injury was treated by continuous cryotherapy with ice bag at 4 °C; the model group was not treated. Histological changes were observed at 48 hours. Injury degree was evaluated using injury symptom index.RESULTS AND CONCLUSION: Compared with model group, the scores of injury symptom index and histology were lower, interleukin-1β expression was reduced, and transforming growth factor-β1 (TGF-β1) expression was increased in intermittent cryotherapy and continuous cryotherapy groups (P < 0.05). Compared with intermittent cryotherapy group, the scores of injury symptom index and histology were reduced (P < 0.05), interleukin-1β expression was reduced (P < 0.05), and TGF-β1 expression was increased in continuous cryotherapy group (P < 0.05). Results demonstrated that cryotherapy can cure the acute damage of soft tissue by reducing interleukin-1β expression and raising TGF-β1 expression. Continuous cryotherapy is superior over intermittent cryotherapy.

Objective To investigate the real type of the first earlier onset spinocerebellar ataxia family in China. Methods Two family members were subjected to autopsy, whose genetypings were confirmed by polymerase chain reaction (PCR) and direct sequencing technique. Golgi staining, immunohistochemistry and electron microscopy methods were used to detect the neurodegeneration in central nervous system of 2 patients. Results The light microscopic and electron microscopic showed synaptic degeneration of Purkinje cell in the cerebellar cortex, which was accompanied by deterioration of Purkinje cell, and both inferior olivary complex, dentate nucleus and anterior central gyrus. Conclusions There is severer neurodegeneration in the central nervous system of earlier onset spiuocerebellar ataxia 6 patient, especially in cerebellar cortex, inferior olivary complex and dentate nucleus, and the neurodegeneration may depend on disease duration.

BACKGROUND:Using three dimensional (3D) reconstruction techniques,any part of body is accessible to visual observation.However,reports concerning 3D reconstruction of craniofacial vascularity based on PC are few in China.OBJECTIVE:To explore the method and the application values of reconstructing a digital 3D model of craniofacial vascularity based on the data of CT strengthening scanning.METHODS:CT strengthening scan images from a healthy volunteer in DICOM format were imported into Mimics 10.01 software and craniofacial blood vessels were reconstructed with the technique of thresholding,editing and 3D region growing.RESULTS AND CONCLUSION:The 3D digital model of craniofacial blood vessels was obtained.This model could be zoomed and rotated randomly and displayed the spatial positions and adjacent relationships of different anatomical structures,also the reconstructed structures could be measured in 3D space.The 3D digital model of craniofacial blood vessels can be reconstructed conveniently and quickly with Mimics software on PC,and also it can bring morphological reference to human anatomy teaching,clinical neurosurgery,oral and maxillofacial surgery,as well as image diagnosis,and will be helpful to generate a virtual plateform in craniofacial surgery.

Anatomic structure in craniofacial region is very complicated and orthognathic surgery is difficultly performed.Therefore,it is necessary to generate virtual surgical model from CT scan data before operation on personal computer in order to make surgery more accurate.This technique has been increasingly interested in the field of orthognathic surgery both at home and abroad.The sample was scanned with thin-layer CT.CT image data (336 layer,1.0 mm slice thickness) were analyzed with the Mimics software.Simultaneously,three-dimensional model and virtual surgery of the facial cranium were established.The digitized virtual surgical platform of facial cranium was preliminarily created,and three common surgical methods in maxillofacial surgery,i.e.,LeFort Ⅰ osteotomy,mandibular angle osteotomy,and genioplasty,for maxillofacial surgery were simulated.The results showed that the digitized virtual surgical platform of facial cranium could be generated with the Mimics software on personal computer,which provides convenient and quick methods for research,teaching,and clinical surgery.More importantly,it creates theoretical basis for virtual surgical platform which can be widely used on personal computer.

Objective To investigate the effects of lipopolysaccharide(LPS) on myelin phagocytosis during Wallerian degeneration after early peripheral nerve injury in rats.Methods Fifty male Wistar rats were recruited and randomly divided into LPS group(n=20),model group(n=20) and sham group(n=10).The right sciatic nerves of rats in the LPS and model groups were cut and sutured end-to-end,while the sciatic nerve of sham group rats were only exposed.Immediately after surgery,the rats in LPS group were given microinjections of LPS(2 g/L) into the surgical site in a final volume of 1 μL,and the rats in other two groups were injected with the same volume of saline.The sciatic nerves were taken at 1.5 h,24 h and 7d after surgery.Real-time quantitative PCR(qRT-PCR) was applied to detect the dynamic expressions of IL-1β mRNA and MCP-1 mRNA.Immunofluorescence staining was used to test the expression of CD68+ macrophages in sciatic nerves.HE staining was used to observe the pathological alterations of sciatic nerves tissue.ORO staining was used to observe sciatic nerves demyelination.LFB staining was used to detect the sciatic nerves myelin.Sciatic function index was used to evaluate the recovery of motor function in rats.Results Compared with the model group,qRT-PCR indicated that the expression of IL-1β and MCP-1 from LPS group were increased at 1.5 h and 24 h after surgery(P<0.001, P<0.001),respectively.Compared with the model group,the expression of CD68+ cells was increased significantly at 7th day after surgery(P<0.05).Histological examination showed that compared with the model group,a lot of inflammatory cells and Schwann cells were found at sciatic nerve stump in the LPS group at 7th day after operation.ORO staining showed that the degree of demyelination in the LPS group was higher than that in the model group.LFB staining showed that the sciatic nerve stump demyelination appeared in both model group and the LPS group at 7th day after operation,but compared with the model group,myelin debris clearance in the LPS group was significantly accelerated(P<0.05).Finally,compared with the model group,the SFI in the LPS group was increased significantly at 20 d after surgery(P<0.05).Conclusions The results confirm that LPS is possible to manipulate the innate immune response to accelerate myelin clearance during Wallerian degeneration after early peripheral nerve injury in rats.

Objective:To investigate the effects of chitin on atopic dermatitis in an OVA induced AD murine model.Methods:Twenty-eight BALB/c mice were randomly divided into three groups:the normal control group (N)(8),the chitin group(E) (10) and the AD model group(M)(10).The murine model of atopic dermatitis was established through intraperitoneal injection of OVA followed by repeated epicutaneous application of OVA on mice back skin( AD model group).During the set up of AD murine model,mice of the chitin group were given intragastric gavage of 3 mg/d for 4 weeks.At the end of the experiment, the mice were sacrificed and skin lesions were biopsied for histological study.HE and O-toluidine stained paraffin sections were observed under microscope.The spleen cells were cultured and challenged with OVA and chitin,respectively,the supernatant was obtained for cytokine determination.Serum levels of total and OVA-specific IgE and total IgG2a were determined with ELISA.Results:Chitin significantly inhibited skin inflammation induced by OVA.Compared with the AD model group,the thickness of the epidermis and dermis in the chitin group were obviously decreased.The numbers of dermal infiltrated inflammatory cells,eosinophils and mast cells were significantly decreased in the chitin group compared with the AD model group ( P<0.05-0.001 ).The serum level of total IgE and OVA-specific IgE were significantly lower in the chitin group than in the AD model group(P<0.05-0.001),while the serum level of IgG2a in the chitin group was significantly higher than that of the AD model group( P<0.001).The cultured spleen cells of the chitin group produced significantly higher levels of IL-12 and IFN-γ,but lower level of IL-4 compared with those of the AD model group after OVA challenge (P<0.05).Conclusion:Chitin can inhibit the inflammation and decease the seum level of IgE in the murine AD model.The antiallergic effect of chitin might be associated with the induced production of Th1 type cytokines by mice spleen cells.

BACKGROUND:The mechanism underlying Wal erian degeneration fol owing peripheral nerve injury is complex. Immune regulation on Wal erian degeneration is beneficial for early repair of perpheral nerve injury.
OBJECTIVE:To investigate the effects of Tol-like receptor 4 (TLR4) antagonist on Wal erian degeneration and axonal regeneration after early peripheral nerve injury in rats.
METHODS:Fifty male Wistar rats were recruited and randomly divided into treatment group (n=20), model group (n=20) and sham group (n=10). The right sciatic nerves of rats in treatment and model groups were cut and sutured end-to-end, while the sciatic nerves of rats in sham group were only exposed. In the treatment group rats were intravenously injected with 0.15 mg/kg TAK-242 via tail vein 1 hour preoperatively and 7 days postoperatively, and the rats in the other two groups were given intravenous injection of the same volume of normal saline. The sciatic nerves were removed at 24 hours, 3, 4 and 7 days after surgery.
RESULTS AND CONCLUSION:Real-time PCR indicated that the mRNA expressions of interleukin-1βand monocyte chemoattractant-1 were significantly increased in the model group compared with the sham group at 24 hours after surgery (both P<0.001), while the expressions were significantly decreased after TAK-242 injection (both P<0.001). Immunofluorescence showed that compared with the model group, down-regulated expression of CD68+and iba1+cel s appeared in the treatment group at 3 days after surgery (P<0.01, P<0.05). Luxol fast blue staining revealed that demyelination at the sciatic nerve stump appeared in both model and treatment groups at postoperative 7 days, but myelin debris clearance in the treatment group was significantly reduced compared with the model group (P<0.05). Hematoxylin-eosin staining showed that a lot of inflammatory cel s, Schwann cells and regenerated nerve fibers at the sciatic nerve stump were found in the model group, while there were few inflammatory cells, Schwann cel s and regenerated nerve fibers in the treatment group at 7 days after surgery. Immunohistochemistry found that the expression of growth-associated protein-43 in the treatment group was significantly lower than that in the model group at 4 days postoperatively (P<0.05). Besides, compared with the model group, a significantly decreased sciatic functional index was found in the treatment group at 20, 30 and 40 days after surgery (P<0.05). These results show that TLR4 antagonists delay early nerve regeneration in rats after sciatic nerve injury probably by inhibiting the TLR4 signaling pathway.

Objective: To study the segment of liver according to the large amount of three-dimensional(3D) reconstructive images of normal human livers and the vascular system, and to recognize the basic functional liver unit based on the anatomic features of the intrahepatic portal veins. Methods: The enhanced CT primitive DICOM files of 1 260 normal human livers from different age groups who treated from October 2013 to February 2017 provided by 16 hospitals were analyzed using the computer-aided surgery system.The 3D liver and liver vascular system were reconstructed, and the digital liver 3D model was established.The vascular morphology, anatomical features, and anatomical distributions of intrahepatic portal veins were statistically analyzed. Results: The digital liver model obtained from the 3D reconstruction of CAS displayed clear intrahepatic portal vein vessels of level four.Perform a digital liver segments study based on the analysis of level four vascular distribution areas.As the less anatomical variation of left hepatic portal vein, the liver was classified into four types of liver segmentation mainly based on right hepatic portal vein.Type A was similar to Couinaud or Cho′s segmentation, containing 8 segments(537 cases, 42.62%). Type B contained 9 segments as there are three ramifications of right-anterior portal vein(464 cases, 36.82%). The main difference for Type C was the variation of right-posterior portal vein which was sector shape(102 cases, 8.10%). Type D contained the cases with special portal vein variations, which needs three-dimensional simulation to design individualized liver resection plan(157 cases, 12.46%). These results showed that there was no significant difference in liver segmental typing between genders(χ²=2.179, P=0.536) and did not reveal any significant difference in liver segmental typing among the different age groups(χ²=0.357, P=0.949). Conclusions The 3D digital liver model can demonstrate the true 3D anatomical structures, and its spatial vascular variations.The observation of anatomic features, distribution areas of intrahepatic portal veins and individualized liver segmentation achieved via digital medical 3D visualization technology is of great value for understand the complexity of liver anatomy and to guide the precise hepatectomy.