Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

BACKGROUND:Fentanyl transdermal system has analgesic effect similar to oral sustained-release morphine and has been widely used in advanced cancer pain management in several years. However, recent literatures about some serious adverse events associated with fentanyl transdermal system have been published, and the long-term safety of fentanyl transdermal system treatment is stil chalenged. OBJECTIVE:To observe the long-term clinical efficacy and safety of fentanyl transdermal system for pain management in patients with advanced cancer. METHODS:A total of 309 patients with advanced cancer pain were enroled, including 166 females and 143 males. The age ranged from 26-72 years old. Patients received oral sustained-release morphine for 2 weeks, and then were subject to fentanyl transdermal system for pain management at the 3rd week until the 12th week. A prospective study with self-contrast method was conducted to compare the analgesic effects of these two drugs, as wel as patient's acceptability, adverse events and toxicity in the administration course. RESULTS AND CONCLUSION: Stable pain relief was harvested throughout the oral administration of both sustained-release morphine and fentanyl transdermal system therapy. During the use of oral sustained-release morphine, adverse reactions appearing in sequence were constipation, nausea, fatigue and anorexia. After converting to fentanyl transdermal system, the symptoms of constipation (χ2=5.22,P=0.02) and nausea (χ2=4.38, P=0.04) significantly reduced, and vomiting was abated but showed no significant difference (χ2=2.7,P=0.10). 2.3% of patients had skin reactions to the patches, and regressed after replacing the patch area. Skin reactions were aleviated at 2-10 weeks after oral administration of fentanyl transdermal system. Some uncommon adverse events including headache, diarrhea, dyspnoea, excessive sweating or other symptoms often occurred at the time of the initial dosage increase. Preference or strong preference for fentanyl transdermal system in comparison to previous oral sustained-release morphine was reported by 91% of patients. These results demonstrate that fentanyl transdermal patches can provide stable pain relief for advanced cancer patients with good acceptability, and mitigate the incidence of adverse events due to oral drugs.

Objective To investigate prevention and treatment of cerebrospinal fluid leakage (CSFL)in anterior cervical spine surgery for severe ossification of posterior longitudinal ligament (OPLL).Methods A retrospective analysis of 47 patients with severe cervical OPLL (thickness of the ossified mass ＞ 5 mm,spinal stenosis ＞50％),who had undergone anterior cervical surgery between January 2008 and May 2011,was conducted.Fifteen cases of dural defect were found intraoperatively,including 11 males and 4 females,aged from 40 to 68 years (average,55.6 years).Preoperative CT scans were earefully analyzed.During the operation,the ossified mass was excised or floated and the arachnoid was reserved in order to reduce dural damage.Dural defects were repaired by suturing or covering with muscle.After surgery,patients were confined to bed rest to allow for drainage or puncture.Results All 15 patients were followed up for 12 to 18months (average,14.8 months).After operation,10 patients recovered fully without CSFL.Five patients developed CSFL,including 4 cases of spinal dural mater injury that healed within 4-6 days with bed rest and pressure dressing,and 1 case of cerebrospinal fluid pseudocyst that disappeared within 3 weeks with repeat puncture and aspiration treatment.No cases required secondary operations or shunt placement.All cases exhibited good neurological improvement.Conclusion During anterior surgical treatment for severe cervical OPLL,CSFL can be effectively prevented through eareful analysis of preoperative CT images,meticulous operative technique,reasonable handling of the ossified mass and positive repair of dural damage.Moreover,postoperative bed rest and drainage are effective to treat CSFL.

Objective To evaluate the clinical outcomes of modified eggshell technique through posterior approach for the treatment of hard thoracic disc herniation. Methods From January 2006 to June 2009, 22 patients admitted for hard thoracic disc herniation were reviewed, including 13 males and 9 females, with an average age of 49.5 years (range, 33-69). The courses of disease were 11 months on the average (range, 3-18 ). The lesions located in T8-9 for 4 cases, T9-10 for 9, T10-11 for 7, T11-12 for 2. Each of the patients underwent X-ray, CT scanning and MRI examination before surgery. There were 16 cases of central type, and 6 cases of paracentral type. All patients were treated surgically by modified eggshell technique via posterior approach. Results The mean operative time was 210 min (range, 180-300 min), with a mean blood loss of 860 ml (range, 600-1200 ml). All surgeries were performed successfully without neurological symptoms aggravation. Surgical complications included dural laceration in 2 cases, both dural lacerations were repaired intraoperatively, epidural hematoma in 1 case with lower extremity neurological symptoms, full neurologic recovery was observed after surgical removal of the hematoma. All patients were followed up for average 27.5 months (range, 12-54). The mean JOA score increased from 3.36±1.79 before operation to 7.45±2.99 after the operation at 12 months follow up, and the mean improvement rate of neurological status was 58.3%±30.7%. There was significant difference in JOA score before and after surgery(t=10. 12,P＜0.01 ).The results of 14 cases were ranked as good, 6 as fair, 2 as unchanged, and none as worsened. All cases obtained bony fusion without instrument failure. Conclusion Modified eggshell technique enable ventral and dorsal spinal decompression from the posterior approach in cases of hard thoracic disc herniation, with reduction of the rate of postoperative paralysis.

Objective To investigate whether increased signal intensity (ISI) can help assess the prognosis in patients with cervical spondylotic myelopathy (CSM) by means of measuring the ratio of signal intensity. Methods A retrospective study with two or more years follow-up of 57 patients with CSM underwent posterior cervical decompression were carried out from February 2000 to February 2006. 1.5T MRI was performed in all patients before surgery. T2-weighted images (T2WI) of sagittal ISI on the cervical spinal cord were obtained, For those with ISI, the values of signal intensity of the spinal cord on T2-weighted image (T2Wl) and TI-weighted image (TIWI) of sagittal view were measured at the location where there was ISI on T2WI, and the ratio of signal intensity of T2WI / T1WI (T2/T1 ratio) at the same level of the spinal cord and with similar area was calculated on the computer. Patients with ISI were subdivided into 2 groups according to T2/T1 ratio. Results ISI was not observed in 20 patients (group 1). The range of T2/T1 ratio of other 37 patients was from 1.28 to 2.80 and the median was 1.65. Nineteen patients were divided into group 2 (ratio range, 1.28-1.63), and 18 into group 3 (ratio range, 1.67-2.80). Significant differences were noted in age at surgery, duration of disease, recovery rate, pre and preoperative JOA score among three different groups.Spearman's rank correlation showed that T2/T1 ratio was positively correlated with age at surgery and duration of disease, negatively with pre- and postoperative JOA score and recovery rate. Conclusion Patients with ISI and higher T2/T1 ratio tend to have relatively severe preoperative state of illness and poor prognosis after surgical intervention. Spinal cord signal intensity change on T2-weighted MRI might be a predictor of a poor outcome in terms of functional recovery rate in patients underwent operations for multi-level CSM.

Objective To evaluate the effectiveness of rehabilitative treatment after extended open-doorlaminoplasty of the cervical spine for cervical spondylotic myelopathy.Methods Sixty patients(mean age 61.7 ±12 years)who had undergone extended open-door laminoplasty of the cervical spine for cervical spondylotic myelopa-thy were studied for an average of 40.7 months.They are divided into a rehabilitation group and a control group with 30 patients in each.The patients in the rehabilitation group received systematic rehabilitation therapy pre-and post-operation.In both groups,the neurological recovery rate,the cross-sectional areas of the cervical posterior muscles,and the incidence of axial symptoms and post-operative complications were recorded and compared. Results The wounds of patients in the rehabilitation treatment group healed with no complicating infections,but the wounds of three patients in the control group became infected.There was no statistically significant difference in neurological recovery between the two groups.In the rehabilitation treatment group,the rate of atrophy and the cross-sectional areas of the cervical posterior muscles were significantly lower than in the control group.At the same time,the rate of neck axial symptoms was 23% in the rehabilitation group but 60%in the control group.a difference which was statistically significant. Conclusions Although rehabilitation treatment pre-and post-operation may not speed up neurological recovery,it may prevent postoperative atrophy of the cervical extensors and lower the incidence of neck axial symptoms.

This study was aimed to establish the three-dimensional model of C_(3-7) segment of lower cervical spine after artificial disc implantation, to analyze the movement of lower cervical spine after artificial disc replacement. According to CT films of 1 patient at 6 months after artificial disc implantation, three-dimensional finite element model that included Bryan~(TM) artificial cervical disc prosthesis of the lower cervical spine was established using finite element method, then introduced into Ansys 9.0, the vertebral cortical bone, cancellous bone and intervertebral disc were meshed and analyzed by using under several states such as flexion/extension, lateral bending and rotation, thus understanding their motion characteristics. By comparison with previous research findings, test results nearly accorded with or exhibited identical trend with previous study. The results suggest that, cervical disc replacement can basically guarantee the stability of lower cervical spine movement.

Granulicatella species are facultative anaerobic, catalase-negative Gram-positive cocci, oral microbiome researches find out Granulicatella species are dominant bacteria in oral cavity which may cause opportunistic infection like periodontal disease, endodontic infection. This review summarized research progress of Granulicatella species.
Bacteria ; Humans ; Mouth ; microbiology

OBJECTIVETo study the effects of inoculum, various media, pH value of medium and illumination conditions on the growth of Panax quinquefolium suspension cells and the synthesis of ginsenosides Re, Rb1 and polysaccharides.

METHODThe suspension cells were obtained through tissue culture by manipulation of inoculum, various media, pH value, and illumination conditions. The contents of ginsenosides Re and Rb1 were determined by HPLC, while the contents of polysaccharide were determined by ultraviolet spectrophotometry.

RESULTThe growth rate of suspension cells was greatly increased when inoculum amount was 25 g x L(-1). The effect of media MS, SH and B5 on suspension cells was observed. MS medium was favorable for cells growth, while B5 medium was favorable for the synthesis of ginsenosides and polysaccharides. The polysaccharide content in three media were higher than that of the cultivations. The pH value showed little influence on the cells growth, medium pH 6.0 enhanced the synthesis of Re and polysaccharides. Illumination could significantly enhance secondary metabolite biosynthesis of suspension cells and promoted slightly in polysaccharide synthesis.

CONCLUSIONThe inoculum, various media, pH value of medium and illumination conditions have significant influences on suspension cells growth of P. quinquefolium, secondary metabolite and polysaccharides synthesis.

Biomass ; Cell Culture Techniques ; Clinical Laboratory Techniques ; Culture Media ; Ginsenosides ; metabolism ; Panax ; chemistry ; growth & development ; metabolism ; Polysaccharides ; metabolism ; Suspensions ; analysis ; chemistry

BACKGROUND: Degenerative lumbar spinal stenosis could be treated by laminectomy internal fixation placement,unilateral or bilateral decompression,posterior laminectomy and so on.However,whether laminectomy internal fixation placement can be used remains unclear.OBJECTIVE: To evaluate the efficacy of interbody fusion cage implantation of pedicle screw fixation,in combination with posterior lumbar laminectomy,bilateral resection and decompression of the inferior articular process,autologous facet joint bone transplantation in the treatment of degenerative lumbar spinal stenosis.METHODS: A total of 41 patients of degenerative lumbar spinal stenosis,who failed after 3 months of conservative therapy,including 23 males and 18 females,at a mean of 60.3 years,Received posterior lumbar laminectomy,bilateral inferior articular process resection and decompression,autologous facet joint bone and cage interbody fusion implant pedicle fixation.They were followed up for 24 months,preoperative and postoperative Japanese Orthopedic Association(JOA)score evaluations were performed to assess the therapeutic efficacy of the patients,radiological examination was done to investigate the graft fusion and vertebral stability of surgical segments in patients.RESULTS AND CONCLUSION: During the follow-up,JOA score significantly increased compared with the preoperative score(P＜0.01)and clinical excellence rate was 90%; 40 cases obtained bony fusion,with a fusion rate of 98%,1 patient exhibited signs of lumbar instability.There was no loosening,fracture and other complications after internal fixation,but 2 cases appeared dural tear,1 case pedicle position deviation,1 case pseudoarticulation formation.The results suggest that the posterior lumbar laminectomy,bilateral inferior articular process resection and decompression,autogenous facet joint bone and cage interbody fusion implanted pedicle screw fixation show good clinical effects for the treatment of degenerative lumbar spinal stenosis.