BACKGROUND: Myelination following axonal regeneration is a key factor affecting the recovery of spinal cord injury. Oligodendrocyte survival directly affects the myelination following axonal regeneration. OBJECTIVE: To investigate the feasibility of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into oligodendrocytes induced by neurotrophic factors. DESIGN, TIME AND SETTING: The cell molecular biology in vitro study was performed at the Laboratory of Department of Orthopaedics, Tongji Hospital from September 2006 to June 2007. MATERIALS: A total of 5 Sprague Dawley rats aged 2-4 weeks, of both gender were selected. Bilateral femur and tibia bone marrow was obtained to harvest BMSCs. METHODS: At passage 4, BMSCs were incubated in serum-free medium, supplemented with N2, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor for 48 hours, and incubated in medium containing 500 ng/mL insulin-like growth factor I and N2 for 3 days. MAIN OUTCOME MEASURES: Morphological changes were observed using an phase contrast microscope. Semiquantitative RT-PCR was utilized to detect specific marker mRNA expression of oligodendrocytes. Using neuron marker anti-microtubule-associated protein, astrocyte marker anti-glial fibrillary acidic protein, oligodendrocyte marker anti-galactocerebroside, anti-myelin basic protein antibody, immunocytochemical staining was performed to detect the positive rate of the differentiation of BMSCs into oligodendrocytes. RESULTS: Morphological changes in BMSCs during the differentiation into oligodendrocytes: After the induction, a majority of BMSCs presented the morphological characteristics of oligodendrocytes. Cytoplasm retraction towards nucleus, cell process extension towards outwards, and strong refraction were found. With the prolongation of time, several cell processes connected and formed a typical net-shape structure. Specific marker mRNA expression of oligodendrocytes: Following induction, specific strap of myelin basic protein mRNA and galactocerebroside mRNA could be detected. Positive rate of oligodendrocytes: During induction, the positive rates of galactocarebroside, myelin basic protein and microtubule-associated protein were 65%, 45% and 10%, respectively. CONCLUSION: The combination of epidermal growth factor, basic fibroblast growth factor and insulin-like growth factor can effectively promote the directional differentiation of BMSCs into oligodendrocytes.

Objective To study the effect of stable expression of reconstructed human transforming growth factor-β3 ( hTGF-β3) on proliferation of precartilaginous stem cells ( PSCs) and their differentiation into cartilage chondrocytes.Methods After isolated and purified by immunological microbeads,PSCs of rats were transfected with pcDNA3.1 ( + )-hTGF-β3.MTT reduction assay ( MTT) and flow cytometry were performed to investigate the effect of transduction on proliferation and DNA synthesis.Biosynthesis of hTGF-β3 and expressions of cartilage associated genes and proteins were examined by qRT-PCR,immunohistology and Western blot.Results hTGF-β3 was expressed in PSCs stably.Compared with non-transfection group,PSCs' DNA synthesis level and proliferation rate were significantly increased after transfection.Quantitative real-time PCR and immunological investigation suggested up-regulated expression of specific genes and proteins of chondrocyte and increase of deposition of chondrocyte typical extracellular matrices proteoglycan and collagen type Ⅱ .Conclusions Gene enhanced PSCs can stably express hTGF-β3 protein to promote proliferation of PSCs and induce differentiation of PSCs in to chondrocytes,which provides a fresh approach to cartilage tissue engineering.

Objective To study the biological effects of sinusoidal electromagnetic fields(EMFs)on proliferation and extracellular matrix(ECM)formation by annulus fibrosus(AF)cells in rats.Methods AF cells isolated from rats were randomly divided into a control group and an experimental group.The cells in the experimental group were stimulated with an EMF,while those in the control group were held under the same culture conditions but with no EMF.Flow cytometry and MTT were performed to observe the effects on the ceU cycle and proliferation.Collagen and aggrecan expression were examined after amplification with a reverse transcriptase polymerase chain reaction(RTPCR).Sulfated glycosaminoglycan(sGAG)content wag detected by applying the Alcian blue method. Results AF cell proliferation was not significant until after 4 days of stimulation.Compared with the control group,the expression of type Ⅰ and Ⅱ collagen and Aggrecan were up-regulated,and sGAG content Was increased in the experimental group.Conclusion AF cell proliferation was enhanced by EMF.Gene expression of collagen type Ⅰ and Ⅱ and Aggrecan increased.a8 well as sGAG levels.The results suggest an approach for treating of intervertebral disc degeneration.

ObjectiveTo analyze the surveillance results at the national surveillance spots of brucellosis from 2006 to 2010,to know the epidemic status of national brucellosis,and to provide scientific evidences for evaluating of surveillance quality and formulating of surveillance strategies and measures.Methods The brucellosis surveillance data collected from the Surveillance Reports of Major Infectious Diseases and Vector-Biological Monitoring Report in China from 2006 to 2010,and the National Diseases Surveillance Information Management System were classified and analyzed using two stage clustering statistical analysis.Results According to national routine surveillance,the incidence of brucellosis increased from 1.55 per 100 000 in 2006 to 2.62 per 100 000 in 2010.Reported brucellosis cases were mainly in Inner Mongolia,Shanxi,Heilongjiang and other regions,and reported time was from March to August,which accounting for 79.87％(114 265/143 064).The brucellosis cases were mainly youth,male,farmers and herdsman.In the national surveillance spots of the brucellosis,217 648 cases received epidemic survey,of which 61 905 cases received serological test,the positive rate was 19.66％(12 169/61 905),and 10 318 new cases were found; the highest positive rate of serological test was 41.19％(2757/6694) which was found in Zhalantun of Inner Mongolia; the detection rate of pathogen culture was 2.49％(29/1165); the positive rates of serological test in cattle and sheep were 1.07％(2355/219 352) and 0.93％ (2766/296 176),respectively.The surveillance spots were classified into four grades according to cluster analysis.Conclusions The epidemic continues to rise in human surveillance spots of brucellosis.Although national surveillance spots play an important role in the surveillance of brucellosis,the quality and quantity of surveillance work of national brucellosis surveillance spots should be improved,different management measures should be taken in different surveillance spots according to classified results of the cluster analysis.This study provides some basis to improve the utilization of health resources and the level of Brucella disease prevention and control work.

OBJECTIVETo explore the effect of Tongfu Jinghua Decoction (TJD) on hemodynamics and tissue oxygen metabolism in patients with post-traumatic sepsis shock.

METHODSTotally 60 patients were randomly assigned to the treatment group and the control group, 30 in each group. Patients in the treatment group took TJD or were administered with TJD by nasal feeding in combined with conventional Western medical treatment, while patients in the control group only received conventional Western medical treatment. Changes of each index in hemodynamics and tissue oxygen metabolism were observed before treatment, and at 6, 12, 24, and 48 h after treatment.

RESULTSCompared with before treatment in the same group, hemodynamic changes were significantly improved at each time point in the two groups. All indices of tissue oxygen metabolism at each time point of the two groups were significantly improved, except changes of O2 extraction ratio (ER) after treatment in the control group (P < 0.05, P < 0.01). Compared with the control group in the same period, heart rate (HR), systemic vascular resist- ance (SVR), and cardiac output (CO) were significantly improved with statistical difference (P < 0.05, P < 0.01). Mean arterial pressure (MAP), central venous pressure (CVP), and cardiac index (CI) were significantly improved at 6, 12, and 24 h after treatment with statistical difference (P < 0.05, P < 0.01). Each index of tissue oxygen metabolism in the treatment group were all improved at each time point with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTJD combined with conventional Western medical treatment could quickly improve hemodynamics and tissue oxygen metabolism disorder in patients with septic shock, and its curative effect was superior to that of conventional Western medical treatment alone.

Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Heart Rate ; Hemodynamics ; drug effects ; Humans ; Oxygen ; metabolism ; Sepsis ; Shock, Septic ; drug therapy ; metabolism

Objective To assess the feasibility and clinical results of video-assisted high anterior transcervical approach (Smith-Robinson) in treatment of spinal lesions of the craniovertebral junction. Methods Between April 2007 to October 2009, nineteen consecutive patients with spinal lesions of the craniovertebral junction were included in the study. There were 9 males and 10 females aged from 16 to 62 years old with a mean of 32 years. The primary pathologies included 4 cases with chronic odontiod fracture, 2 cases with purely irreducible atlantoaxial dislocation, 6 cases with os odonteideum, 1 case with Marfan synd rome, 1 case with primary basilar invagination from Kippel-Feil syndrome, 3 case with axis tumor and 1 case with irreducible rheumatoid atlantoaxial dislocation. All of the patients underwent combined video-assisted high anterior transcervical procedure and posterior fixation at one-stage. The anterior procedure included atlantoaxil release and reduction (8 cases), odontoidectomy (8 cases), and intralesional extracapsular excision and reconstruction (3 tumor cases). The posterior technique were C1-C2 pedicle screw fixation (13 cases), C1-C3 pedicle screw fixation (2 cases), and occipitalcervical fusion (4 cases). Results Anatomical reduction was achieved in eight cases with anterior release and reduction. Tumors were completely removed in three cases with axial tumor. The mean follow-up was 14 months (6-36 months). All of them achieved solid bone fusion. In the 14 patients with symptoms of spinal cord dysfunction, the average Japanese Orthopaedic Association (JOA)score had improved from 9.1±3.3 preoperatively to 14.1±2.9 postperatively. The improvement rate was excellent for 7 cases, good for 5 cases, fair for lcase and poor for 1 case. One patient experienced leakage of cerebrospinal fluid which was resolved by bioprotein gelatin blocking and lumbar subarachnoid continuous drainage within 1 week. Dysphagia which occurred in 3 cases responded well to dexamethason and mannitol.No infection and hardware failure were observed. Conclusion Video-assisted high anterior transcervical procedure is a safe and effective alternative for treating spinal lesions in the craniovertebral junction.

OBJECTIVETo observe the protection of Qingyuan Shenghua Decoction (QSD) on multiple organs of sepsis patients after bone trauma, and to preliminarily explore its mechanism.

METHODSTotally 60 sepsis patients after bone trauma were randomly assigned to the treatment group and the control group according to random digit table, 30 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took QSD or were nasally fed with QSD, one dose per day for 1 week. Changes of WBC, oxygenation index (PaO2/FiO2), serum creatinine (SCr), total bilirubin (TBIL), aspartate aminotransferase (AST), fibrinogen (FIB), D-dimer (DD), activated partial thromboplastin time (APTT), pro-calcitonin (PCT), C-reactive protein (CRP), heart rate (HR), mean arterial pressure (MAP), intra-abdominal pressure, scores for Acute Physiology and Chronic Health Evaluation II (APACHE II), sequential organ failure assessment (SOFA) scores were observed before treatment and on day 1, 3 and 7 after treatment.

RESULTSCompared with the control group at the same time point, MAP increased at post-treatment day 1 and 3; CRP, APTT, HR, SCr, TBIL, AST, intra-abdominal pressure at post-treatment day 3 obviously decreased in the treatment group (P < 0.05, P < 0.01). WBC, SOFA scores, PCT, CRP, APACHE II, APTT, D-D, HR, SCr, TBIL, AST and intra-abdominal pressure significantly decreased; FIB, MAP and PaO2/FiO2 obviously increased at post-treatment day 7 (P < 0.05, P < 0.01).

CONCLUSIONQSD had good protective effect on multiple organ function in sepsis patients after bone trauma, and its mechanism might be related with effectively clearing endotoxin, alleviating inflammatory reactions, and fighting against coagulation dysfunction.

APACHE ; Blood Coagulation ; Bone Diseases ; complications ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Inflammation ; Partial Thromboplastin Time ; Protein Precursors ; metabolism ; Sepsis ; drug therapy ; etiology

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the clinicopathological features and behavior of a group of jaw cysts with a solely orthokeratinized lining epithelium.

METHODS20 cases of this cyst type were reported under the term of orthokeratinized odontogenic cyst (OOC) and their clinical, histological and immunocytochemical features were compared with that of odontogenic keratocyst (OKC).

RESULTSThe cysts of the present series were all solitary lesions, occurred mostly in young male patients, and showed a predilection for the posterior mandible areas. Follow-up of 15 patients revealed no recurrence following enucleation. Histological and immunocytochemical studies indicated that OOC epithelium lacked the typical features of OKC and appeared to show a lower proliferative activity.

CONCLUSIONThese findings suggest that OOC is clinicopathologically distinctive from OKC and may thus constitute its own clinical entity.

Adolescent ; Adult ; Aged ; Carcinoembryonic Antigen ; analysis ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Male ; Mandibular Diseases ; metabolism ; pathology ; Middle Aged ; Mucin-1 ; analysis ; Odontogenic Cysts ; metabolism ; pathology

Bodily Secretions ; virology ; Humans ; Pharynx ; virology ; Pneumonia ; diagnosis ; virology ; Respiratory System ; virology