BACKGROUND:Recently,in vitro nucleus gelatinosus and annulus fibrosus cell culture technology,especially cartilage tissue engineering development and primary success in autogenous intervertebral disc cell transplantation to repair nucleus pulposus defect,brings a hope for regeneration and reparation of degenerated intervertebral disc.OBJECTIVE:To explore the dose-dependent and time-dependent effects of insulin-like growth factor-Ⅰ(IGF-Ⅰ) on proliferation of anulus fibrosus cells in vitro.DESIGN,TIME AND SETTING:Single-sample observation was performed at the Laboratory of Parasite,Shanxi Medical University between September 2006 and January 2007.MATERIALS:Thirty 1-month-old Wistar rats,irrespective of gender,were selected.METHODS:Anulus fibrosus cells were isolated and cultured in vitro.In dose-dependent test,IGF-Ⅰ(0.1,1.0,10,and 100 ?g/L),prepared by volume fraction 0.01 or 0.1 calf serum HAMF-12,were added.Cells not with IGF-Ⅰ served as control.Cells were cultured for 72 hours.In time-dependent test,optimal dose of IGF-Ⅰ containing volume fraction 0.1 calf serum and F-12 solution was added,and cells not with IGF-Ⅰ served as control.The cells were cultured for 1,3,5,and 7 days.MAIN OUTCOME MEASURES:Cell biological features were detected using HE staining and toluidine blue immunohistochemical staining;Dose-dependent and time-dependent effects were examined by MTT assay.RESULTS:HE stained cells were fusiform-shaped with pseudopodia,round or oval nucleus.The cytoplasm was blue after stained with the toluidine blue.The immunohistochemical test revealed that there existed positive expression of typeⅠcollagen in the cells.In the presence of 10% calf serum,IGF-Ⅰ significantly improved cell proliferative activity in a dose-dependent manner within effective dose range.CONCLUSION:IGF-Ⅰ can stimulate cell proliferation of rat anulus fibrosus cells in vitro in a dose-dependent and time-dependent manner.

Objective To discuss clinical features and surgical treatment of the coexistence of cervical,thoracic and lumber degenerative disease.Methods From January 2004 to December 2008,79 cases with the coexistence of cervical,thoracic and lumber degenerative disease were surgically treated,including 51 males and 28 females,aged from 30 to 80 years(average,58.1 years).All 79 cases presented coexistence of cervical,thoracic and lumbar compression symptoms,such as limb numbness,weakness,zonesthesia and positive Hoffmann and Babinski sign.Forty-one cases mainly presented cervical symptoms,5 presented thoracic symptoms and 12 presented lumbar symptoms.According to affected segments,41 cases received cervical operations,5 cases received thoracic operations,12 cases received lumber operations,and 21 cases received two parts operations.Surgical outcomes were evaluated by Japanese Orthopaedic Association(JOA)score.Results JOA score of all cases improved in varying degrees.The improvement rate of received cervical operation was 66.06%±14.33%,and that of received mere thoracic operation was 56.19%±9.85%,and that of lumbar operation was 63.49%±9.78%.Twenty-one cases received tow parts operations.Forteen cases received one-stage two parts operations with improvement rate of 76.78%±3.94% and 7 cases received twostage two part operations with improvement rate of 71.79%±8.74%.Conclusion The coexistence of cervical,thoracic and lumber degenerative disease is a kind of complex disease,which involves multi-segments and presents various clinical manifestations.The condition was caused by disk herniation,spinal stenosis and ossification of posterior longitudinal ligament and hypertrophy or ossification of ligamentum flavum.The optimal surgical procedures should be selected by integrated judgment of symptoms and imaging manifestations.

Objective To investigate reasonable surgical approaches for lower spine injuries.Methods The study involved 174 patients with lower cervical spine injuries treated with anterior approaches, posterior approaches, or anterior-posterior approaches in our hospital from August 2005 to September 2009. American Spinal Injury Association (ASIA) grading system was used to evaluate the surgical outcome. Results All patients were followed up for average 30 months (6-55 months), which showed that bone union was achieved in 169 patients, with no breakage, loosening or displacement of the internal fixators. There were five deaths. The ASIA grades of 125 patients were improved by 1 or 2 levels ( 1.12levels on average), with an improvement rate of 71.3%. Conclusion Comprehensive and accurate preoperative diagnosis is the basis for choose of correct surgical approaches for lower spine injury. Early and correct surgery is essential for a good prognosis.

BACKGROUND:With the development of the research, induced pluripotent stem cel s are applied to the build of disease model, drug screening, regenerative medicine, and many other research fields, and have made significant achievements, especial y in the study of nervous system diseases.
OBJECTIVE:To summarize the recent development of induced pluripotent stem cel s and to raise problems and prospects based on the latest research in this field.
METHODS:The first author searched the PubMed database for articles about the induced pluripotent stem cel s, including reviews, clinical research and basic research, published from January 2006 to September 2014. The keywords were“iPS, induced pluripotent stem cel”, and final y 60 articles were included in result analysis.
RESULTS AND CONCLUSION:Induced pluripotent stem cel research continues to make breakthrough from its discovery by Yamanaka’s team in 2006 to winning Nobel Prize in 2012. Induced pluripotent stem cel research has broad prospects in the disease model construction, drug screening and regenerative medicine. Currently, problems such as reprogramming methods, cel stability, and clinical transformation stil need to be solved, and further researches are necessary.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology

Acanthosis Nigricans ; diagnosis ; genetics ; pathology ; Adolescent ; Antigens, CD ; genetics ; Biomarkers ; blood ; DNA Mutational Analysis ; Donohue Syndrome ; diagnosis ; genetics ; pathology ; Humans ; Insulin Resistance ; genetics ; Male ; Mutation ; genetics ; Polymerase Chain Reaction ; Receptor, Insulin ; genetics ; Sequence Analysis, DNA

BACKGROUND: Human umbilical cord Wharton's jelly-derived mesenchymal stem cells (hWJCs) via transplantation are able to survive in a dog model of intervertebral disc degeneration and to partially restore the height of the intervertebral disc. However, there is no study on whether hWJCs can repair nucleus pulposus cells after inflammatory injury as yet. OBJECTIVE: To observe the protective effect of hWJCs on interleukin-1β induced rat nucleus pulposus degeneration.METHODS: Nucleus pulposus cells at passage 3 from Sprague-Dawley rats were divided into three groups: (1) normal control group cultured in culture medium containing fetal bovine serum for 24 hours, and then cultured in complete medium for 24 hours; (2) intervention group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for 24 hours; (3) co-culture group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for non-direct co-culture with hWJCs for 24 hours. Annexin V-FITC/propidium iodide (PI) double staining and flow cytometry were used in succession to measure the apoptosis rate of nucleus pulposus cells in each group. The ADAMTS-4, MMPs-3 and TIMP-1 expression in nucleus pulposus cells was observed though RT-PCR. The activities of caspase-3, caspase-8 and caspase-9 were determined by caspase kit.RESULTS AND CONCLUSION: The apoptosis rates were ranked as follows: the intervention group > co-culture group > normal control group, and there were significant differences between groups (P < 0.05). Compared with the control group,the expression levels of ADAMTS-4 and MMPs-3 were significantly higher and the expression level of TIMP-1 was significantly lower in the intervention and co-culture groups (P < 0.05). Compared with the intervention group, the expression levels of ADAMTS-4 and MMPs-3 were significantly lower and the expression level of TIMP-1 was significantly higher in the co-culture group (P < 0.05). The expression activities of caspase-3, caspase-8 and caspase-9 were highest in the intervention group, followed by the co-culture group, and lowest in the normal control group, and there were significant differences between groups (P < 0.05). To conclude, hWJCs has obviously inhibitory effect on the apoptosis of degenerated nucleus pulposus cells induced by interleukin-1β.

Objective To investigate the distribution and characteristics of drug resistance of pathogens in premature infants with ventilator‐associated pneumonia(VAP) ,and explore the therapeutic measures for premature infants with VAP ,so as to provide references for clinically reasonable administration of antibacterial agents .Methods A total of 54 cases of premature infants diag‐nosed with VAP in the First Affiliated Hospital of Xiamen University from January 2013 to June 2014 were enrolled in this study . Specimens of respiratory tract secretion were collected ,and species identification of pathogens and drug sensitivity test were per‐formed by VITEK 2 Compact system .Results A total of 69 strains were isolated .Gram negative bacteria was accounted for 94 .20% ,and gram positive bacteria was accounted for 5 .80% .Four of the most common pathogenic bacteria were Klebsiella pneu‐monia (29 strains ,42 .03% ) ,Enterobacter aerogenes (12 strains ,17 .39% ) ,Pseudomonas aeruginosa (10 strains ,14 .49% ) and Stenotrophomonas maltophilia(7 strains ,10 .14% ) .The results of antibiotics‐sensitivity tests showed that these strains were multi‐drug resistant to most commonly used antibiotics ,but sensitive to only a few antibiotics .Conclusion Gram‐negative bacteria are the major pathogens for premature infants with VAP in this hospital .Mixed infection and multidrug resistance are common .Therefore , the rational use of antibiotics according to drug sensitivity tests′results is critical in treating VAP .

Objective: To compare trophic effects of soluble substances derived from Schwann cells (SC) of neonatal and Wallerian degenerating segments of rats on facial motoneuron (FMN) cultures. Methods: Serum-free conditioned media of Schwann cell cultures (SC-CM) from facial and sciatic nerves of neonatal and Wallerian degenerating segments in adult rats were individually collected and concentrated by ultra-filtration with molecular weight cut-off at 30 000 and 10 000. The growth activities of FMNs in vitro were determined by means of MTT assay under the condition of serum-free medium added with different components of concentrated SC-CMs (SC-CMCs). The absorbance values were then statistically analyzed. Results: Survival and growth rate of FMN cultures in four kinds of SC-CMCs were significantly higher than that in media both with serum and non-serum and the difference between SC-CMCs was not statistically significant (P>0.05). Neurotrophic molecules were predominantly protein or peptide components with relative molecular weight larger than 30 000 and their trophic activity was positively related to total protein concentration in SC-CMCs. Conclusion: There were soluble trophic molecules with relative molecular weight larger than 30 000 for survival and neurite growth of FMN cultures in media with SC-CMCs derived from facial and sciatic nerves of neonatal rats and with SC-CMCs derived from Wallerian degenerating facial and sciatic nerves of adult rats. It might be reasonable to choose SC from sciatic nerves of rats on account of the findings from SC cultures on facial motoneurons.

Objective To investigate the clinical efficacy,operative essentials and indications of hybrid spinal fusion surgery for cervical spondylotic myelopathy.Methods From August 2008 to December 2011,thirty-eight patients with cervical spondylotic myelopathy underwent hybrid spinal fusion surgery in our hospital.There were 27 males and 11 females,aged from 33 to 70 years (average,51 years).A total of 86 segments were treated (fusion 48 vs.non-fusion 38).Twenty-eight patients underwent a two-level surgery,and ten patients received a three-level surgery.The Japanese Orthopaedic Association (JOA) score and Visual analogue scale (VAS) were used to evaluate pre-and post-operative neurological function and pain,respectively.The pre-and post-operative range of motion of the cervical spine was measured according to Xrays.Moreover,the surgical complications were recorded and analyzed.Results Thirty-seven patients were followed up for 15 to 55 months (average,29.1 months).The improvement of neurological function was obtained in 36 patients.The JOA score was improved from preoperative 10.5±1.57 to 14.3±1.97 at final follow-up,with an improvement rate of 58.46％,and the results were excellent in 16 cases,fair in 20 cases and poor in 1 case.The VAS was improved from preoperative 7.3±1.04 to 3.2±1.41 at final follow-up.The Cobb angle changed from preoperative 25°±3.21°to 20°±2.56°at final follow-up.After operation,the neurological function was not restored in 1 case; hoarseness and bucking occurred in 2 cases; sore throat occurred in 22 cases; anterior displacement of prosthesis (PCM) occurred in 3 cases.Other patients had no complications,such as displacement,loosening and heterotopic ossification.Conclusion In hybrid spinal fusion surgery,the lesions segments are decompressed fully,the severely degenerative segments are fused,and the motion of the non-fusion segments is reserved.As a result,not only the stability of the cervical spine is achieved,but also an obvious improvement of symptoms and a satisfactory short-term efficacy can be obtained.Therefore,this method is an alternative procedure for cervical spondylosis myelopathy.