OBJECTIVETo explore the method for obtaining olfactory ensheathing cells from human fetal olfactory mucosa by cell culture for selective adhesion in the presence of neurotrophin-3 (NT3) and low-concentration serum.

METHODSThe olfactory ensheathing cells were cultured alternatively in DMEM/F12 culture medium containing 10% fetal bovine serum (FBS) and the medium containing NT3 and 2.5% FBS every 72 h. The cells were observed for morphological changes and identified using immunocytochemistry with P75NTR and GFAP, and the cell purity was estimated.

RESULTSThe olfactory ensheathing cells from human fetal olfactory mucosa were positive for P75(NTR) and GFAP, and in in vitro culture, the cells exhibited dipolar or tripolar appearance with long thin neurites. On the 9th day of cell culture, the purity of the olfactory ensheathing cells reached about 83%.

CONCLUSIONThe olfactory ensheathing cells can be obtained by in vitro culture for selective adhesion in the presence of NT3 and low-concentration serum.

Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Culture Media ; Fetus ; Humans ; Neurotrophin 3 ; pharmacology ; Olfactory Bulb ; cytology ; Olfactory Mucosa ; cytology

OBJECTIVETo investigate the expression of telomerase in various lesions of adrenal cortex.

METHODSBy autoradiography-based telomeric repeat amplification protocol, telomerase expression was detected in 36 samples of adrenocortical lesions, including 29 cases adrenocortical adenoma (8 Cushing's syndrome, 17 aldosteronism and 4 nonfunctional adenomas), 5 cases of hyperplasia of adrenal cortex (presented with Chushing' syndrome), 2 cases adrenocortical carcinoma, and 4 samples of normal adrenal cortex.

RESULTSOf the 40 samples, 2 cases of adrenocortical carcinomas had telomerase expression, and the others had no telomerase expression detected.

CONCLUSIONSNo significant telomerase expression was found among different endocrine functional benign adrenocortical lesions. Telomerase expression may be used as an important marker of malignant adrenocortical tumor.

Adrenal Cortex ; enzymology ; Adrenal Cortex Neoplasms ; enzymology ; Adrenocortical Adenoma ; enzymology ; Biomarkers, Tumor ; analysis ; Cushing Syndrome ; enzymology ; Humans ; Telomerase ; analysis ; biosynthesis ; genetics

OBJECTIVETo investigate the changes in the range of motion (ROM) and stress of the intervertebral disc and facet joint of the adjacent segments following anterior cervical corpectomy and fusion (ACCF) and anterior cervical discectomy and fusion (ACDF) using finite element analysis.

METHODSA three-dimensional finite element model of the lower cervical vertebrae was constructed and validated by comparing the ROM of the finite element model against the published data. After the validation of successful modeling, finite element models of ACDF and ACCF were constructed. The ROM and the stress of the intervertebral disc and facet joint of the adjacent segments were compared between the intact lower cervical vertebrae and the cervical vertebrae after ACDF and ACCF.

RESULTSThe ROM of the finite element model was consistent with the published data. The total ROM and the ROM of the fusion segments with ACDF and ACCF were significantly decreased compared with the intact cervical vertebrae. In the adjacent segments following ACDF and ACCF, the ROM the adjacent segments and the stress peak of the intervertebral disc and facet joint all increased obviously compared with those of intact cervical vertebrae.

CONCLUSIONAfter fusion surgeries, the total ROM of the cervical vertebrae decreases and the ROM of the adjacent segment increases. The stress peak of the intervertebral disc and facet joint of the adjacent segments also increases to significantly alter the physiological characteristics of the intact cervical vertebrae.

OBJECTIVETo explore effect and the application value of continuous douche and vacuum sealing drainage (VSD) in refractory tissue, and joint infections after complete debridement.

METHODSAs retrospective analysis of treatment time and restoration or recurrence, from Jan. 2006 to Dec. 2007, 61 cases of refractory tissue, bone and joint infections underwent continuous douche and VSD combined with the treatment of anti-inflammatory and rehabilitation training after debridement in our hospital. The 61 patients included 39 males and 22 females with age ranging from 10 to 58 years with an average of (35 +/- 12) years, among whom 61 identified to have ankle ulcers combined with infections,open fracture combined with infections, sacrococcygeal pressure ulcers combined with infections, infections after hip replacement, infections after open fracture, and infections after skin avulsion postoperation were 11, 15, 9, 3, 5 and 18 cases respectively. The course was from 2 weeks to 11 months with an average of 4 months.

RESULTSIn all 61 patients,the mean healing time was 17, 36, 42, 24, 32, 29 and 28 days in ankle ulcers and infections, tibia and fibula open fracture and infections, femoral shaft fracture and infections, sacrococcygeal pressure ulcers and infections, infections after hip replacement, infections after open fracture, and infections after skin avulsion postoperation respectively. The replacement of VSD was 1, 2-4, 3-5, 1-3, 2-4, 2-3 and 1-3 times in each group respectively. There was no wound recurrence except for 2 cases with recurrent in 61 cases with external fixation nail hole semi-pathological fracture in 1 case of femoral shaft fracture and infection and 1 case of tibia and fibula fracture and infection after follow-up at least one year.

CONCLUSIONApplication of continuous douche and VSD can effectively decrease incidence of complications and promote the refractory tissue, bone and joint infections wound growth, healing and considerably shorten the healing time.

Adolescent ; Adult ; Bone Diseases ; pathology ; physiopathology ; surgery ; Child ; Debridement ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Joint Diseases ; pathology ; physiopathology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Suction ; methods ; Therapeutic Irrigation ; methods ; Time Factors ; Wound Healing ; Young Adult

Objective To explore the clinical efficacy of one-stage anterolateral-posterior approach debridement,bone graft and internal fixation in treatment of thoracolumbar spinal tuberculosis.Methods From January 2010 to December 2014,56 cases of thoracolumbar spinal tuberculosis were retrospectively analyzed, including 31 males and 25 females,aged 18 -72 years (mean 43.1 years).All patients were managed by standard courses of chemotherapy with quadruple anti-TB drugs for 2 - 4 weeks.Patients were treated by anterolateral debridement,autologous iliac bone graft fixed by absorbable screw fixation,and posterior pedicle screw fixation via multi-split muscle gap (Wiltse approach).We recorded the operation time,the amount of bleeding,bone graft fusion,postoperative erythrocyte sedimentation rate (ESR),Cobb angle,VAS score,and American Spinal Injury Association (ASIA)score to evaluate the surgical results.Results The average operation time was 175-290 min, with an average of (248±42)min.The bleeding volume was 300 -900 mL with an average of (420 ±68)mL.The average follow-up time was (24 ± 5.2 )months,bone fusion rate was 100%,and fusion time was (4.7 ± 0.5 ) months.At the last follow-up,the average Cobb angle was (8.2±3.1)°,VAS was (2.1±0.8),and ESR was (17± 4.2)mm/h.The ASIA neurological functions were all classified as Grade E except for 3 cases of Grade D.All these were significantly different from the preoperative ones.Six patients had complications of different degree but without serious complications.Conclusion One-stage anterolateral debridement,autologous iliac bone graft fixedby absorbable screw fixation,and posterior pedicle screw fixation via multi-split muscle gap (Wiltse approach)can completely remove the tuberculosis lesions and achieve ideal kyphosis correction,high bone graft fusion,and satisfactory neurological function recovery.Absorbable screws can be safely applied to the bone graft site after debridement.

OBJECTIVETo retrospectively analyze the surgical methods and its clinical effects and explore a clinical classification and treatment strategy for atlantoaxial dislocation(AAD).

METHODSThe clinical data of 89 patients with atlantoaxial dislocation were analyzed from September 2005 to September 2013. There were 49 males and 40 females, aged from 13 to 67 years with an average of 48.1 years. According to the reductive effects with preoperative cervical dynamic radiograph and high weight skeletal traction under general anesthesia, the dislocations were classified into three types:easy reduction type, hard reduction type and irreducible type. The patients with easy reduction type were treated with posterior screw rod internal fixation after manual reduction, while the patients with hard reduction type were treated with posterior screw rod fixation after high weight skeletal traction reduction under general anesthesia. The patients with irreducible type were treated with transoral atlantoaxial joint release or depression and posterior internal fixation and fusion. According to JOA scores to evaluate the neurological status and treatment outcome.

RESULTSThirty patients were classified as easy reduction type, 55 patients as hard reduction type, and 4 patients as irreducible type. The preoperative JOA score was 8.2±3.1 on average, while the postoperative score was 14.2±2.4. The improvement rate was 40.1% to 82.5% with an average of 62.5%. Eighty nine patients were followed up from 6 to 37 months with a mean of 17.3 months. Eighty two cases obtained anatomical reduction and 85 cases obtained bony fusion. One case complicated with hyponatremia after operation and 1 case combined with Guillain-Barre syndrome, 4 cases complicated with delayed union wounds, 1 case died of for respiratory failure 2 years after operation. No wound infections were found in the patients approach for transoral operation.

CONCLUSIONSAccording to the cervical dynamic radiograph and high weight skeletal traction under general anesthesia to classify for atlantoaxial dislocation, and adopting well strategies to treat the patients, can achieve satisfactory effects.

OBJECTIVE: To screen for aberrantly expressed genes in osteosarcoma cells and investigate the role of RHPN2 in regulating the proliferation, apoptosis, migration and tumorigenic abilities of osteosarcoma cells. METHODS: We used GEO2R to analyze the differential gene expression profile between osteosarcoma cells and normal cells in the GSE70414 dataset. RTqPCR and Western blotting were performed to detect RHPN2 expression in osteosarcoma cell lines MG-63, 143B and SAOS2. Two RHPN2-shRNA and a control NC-shRNA were designed to silence the expression of RHPN2 in 143B cells, and CCK8 assay, colony-forming assay, annexin V-FITC/PI staining and scratch assays were carried out to examine the changes in proliferation, apoptosis and migration of the cells. We also established nude mouse models bearing osteosarcoma xenografts derived 143B cells and RHPN2-shRNA-transfected 143B cells, and assessed the effect of RHPN2 silencing on osteosarcoma cell tumorigenesis using HE staining. Kaplan-Meier survival curves were used to analyze the correlation between RHPN2 expression and survival outcomes of patients with osteosarcoma. RESULTS: RHPN2 expression was significantly upregulated in osteosarcoma cell lines MG-63, 143B and SAOS2 (P < 0.01). Silencing of RHPN2 significantly inhibited the proliferation and migration of 143B cells in vitro, promoted cell apoptosis (P < 0.01), and suppressed tumorigenic capacity of the cells in nude mice. A high expression of RHPN2 was significantly correlated with a poor prognosis of patients with osteosarcoma (P < 0.05). CONCLUSION RHPN2 is highly expressed in osteosarcoma cells to promote cell proliferation and migration and inhibits cell apoptosis. A high expression of RHPN2 is associated with a poorer prognosis of the patients with osteosarcoma.
Adaptor Proteins, Signal Transducing/metabolism* ; Animals ; Apoptosis ; Bone Neoplasms/metabolism* ; Carcinogenesis ; Cell Line, Tumor ; Cell Movement/physiology* ; Cell Proliferation/physiology* ; Humans ; Immediate-Early Proteins ; Mice ; Mice, Nude ; Osteosarcoma/metabolism* ; RNA, Small Interfering/genetics*