BACKGROUND: Although cultured technique and purified method of Schwann cells (SCs) are changed successively,purity is still limited. Whether double negative selections can culture high-purified SCs still needs a further study.OBJECTIVE: To investigate the effects of double negative selections on growth and purity of SCs.DESIGN: Controlled animal study.SETTING: Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University,and the Beijing Institution of Otolaryngology.MATERIALS: The experiment was carried out in the Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University and the Beijing Institution of Otolaryngology between May 2006 and July 2006. Twenty new-born Wistar rats of 3 days (without gender limitation) were provided by the Experimental Animal Center of the Chinese Medical Academy.METHODS: The sciatic nerves of rats were digested by 0.2% collagenase. ① Grouping: All the rats were divided into 4 groups with 5 each. The experimental group was depurated by differential adhesion and Ara-c for 30 minutes. SCs in the control group 1 and 2 were managed by differential adhesion and Ara-c for 30 minutes, respectively. Cells in the control group 3 were not purified. ② Evaluation: The cell growth was observed under invert microscope; growth curves were measured with MTT method in the experimental group and control group 3 to observe cell proliferation. In addition, the SCs were identified by immunocytochemistry; the culture purity was calculated (ratio of S100 positive cells in each sight).MAIN OUTCOME MEASURES: ① Growth of SCs; ② immunocytochemistry stain of SCs; ③ growth curve of SCs; ④ purity of SCs.RESULTS: ① Growth of SCs: At 5 days after incubation, the SCs proliferated well. SCs were spindle, bipolar and sometimes tripolar, with obvious nuclei and a little cytoplasm. There was more fibroblast cell in the group 3 than that in experimental group. ② Growth curve: The SCs in experimental group entered into the logarithm multiplication period at the third day. ③ Immunocytochemistry: The positive-stained SCs were brown in its body and process, and arranged like swirl. The FB with spread and anomaly shape was slight blue. ④ The ratio of anti-S100 positive cells was higher in experimental group than that in group 3 (P ＜ 0.01).CONCLUSION: With the double negative selection, high-purified SCs are obtained effectively.

Objective To investigate the changes of hair growth with the increase of age and to develop a new practical method to evaluate the extent of aging. Methods After the approval of the ethnic committee, 96 healthy volunteers (44 males, 52 females, aged 30 ~ 78 years) were recruited into the study and divided into 4 groups based on the age: 30-group, 40-group, 50-group and 60-group. Macrophotographs were taken from the vertex. Hair density and percentages of black hairs were counted using Photoshop7.0 software. Ten hairs were randomly plucked from the specified scalp area,and the diameters of hair shaft and hair bulb were examined by a micrometer. The percentages of the follicles, based on their morphology, at different phases of the hair cycle, were counted. Results With the increase of age, the hair density, diameters of hair shaft as well as hair bulb, the percentages of anagen follicles and grey hairs all decreased, while the percentages of telogen follicles and white hairs increased, and significant difference was found between the 60-group and other 3 groups. In each group, the difference between males and females was found only in some parameters. The diameter of hair bulb positively correlated with that of hair shaft. Conclusion This study suggests that the parameters of hair growth could be used to help rating the extent of aging.

Objective To determine the in vitro effects of the traditional Chinese medicines, berberine, matrine and baicalin, on the proliferation and lipid synthesis of human immortalized sebocyte SZ95, and to investigate their possible mechanisms of action on sebaceous glands at the cellular level. Methods Inverted microscopy was used to observe cell morphology and determine toxic concentrations of the compounds. The MTT method was adopted to examine the effects of different concentrations of berberine, matrine and baicalin on SZ95 cell proliferation after 24 h, 48 h and 72 h of incubation. Lipid contents in the SZ95 cells were labeled with Nile red and analyzed by flow cytometry. Results The toxic concentrations were 1?10-3 mol/L, 1?10-4 mol/L, 1?10-3 mol/L for baicalin, berberine and matrine, respectively. Berberine reduced sebocyte proliferation in a dose- and time-dependent manner, with an inhibitory concentration 50 (IC50) of 2.9?10-5 mol/L and 1.4?10-5 mol/L after 48 h and 72 h of incubation, respectively. When the concentration of matrine was 0.05). Lipogenesis of SZ95 cells showed a 26.9% increase with 1?10-3 mol/L matrine. Conclusions Our results indicate that berberine and baicalin can inhibit proliferation and lipid synthesis of SZ95 sebocytes in vitro, which suggests a possible clinical role in treating acne.

This study was aimed to observe effects of Albizia Julibrissin Flower Total Flavonoids (AJFTF) on the learning and memory abilities and plasma monoamine neurotransmitters 5-HT and NE content of depression model rats . A total of 90 SD rats were randomly divided into the normal group , model group , western medicine group ( Venlafaxine Hydrochloride 0 . 0125 g?kg-1 ) and high , middle and low dosage groups of AJFTF ( equivalent to the crude drug of 10 , 5 , 2 . 5 g?kg-1 ) . Depression model rats were induced by solitary cultiva-tion and chronic unpredictable stress . The learning and memory abilities of model rats were evaluated with the Morris water maze . And the plasma contents of 5-HT and NE were determined by enzyme-linked immunosor-bent assay ( ELISA ) . The results showed that AJFTF ( administered with high , middle and low dosage for 21 days ) significantly shortened the incubation period of the Morris water maze positioning and navigation ( P <0 . 05 or P < 0 . 01 ) , and increased the times of crossing the platform in space exploration ( P < 0 . 05 or P <0 . 01 ) . There was a certain dose-effect relationship . The AJFTF ( high , middle and low dosage ) increased the plasma contents of 5-HT and NE ( P < 0 . 05 ) . It was concluded that AJFTF can improve learning and memory abilities of depression model rats which indicates that AJFTF have potential antidepressant effects . Its action mechanism may be related to increase plasma monoamine neurotransmitter content of 5-HT and NE .

Objective:To study the relationship between the recurrence of benign meningiomas and cell proliferation, and to find some parameters that can predict the recurrence of benign meningiomas. Methods:49 samples from 15 recurrent benign meningiomas and 22 non recurrent benign meningimas were immunohistochemically stained with PCNA antibody and Ki 67 antibody.The ratios of positive tumor cells were calculated,then PCNA LI and Ki 67 LI of different groups were compared. Results:PCNA LI or Ki 67 LI in recurrent benign meningiomas was significantly higher than non recurent benign meningiomas individually( P 0.05). Conclusion:The proliferative potential of recurrent benign meningiomas was higher than that of non recurrent benign meningiomas.When the PCNA LI was greater than 2.0%,the recurrent risk was high.The proliferative potential did not increase when benign meningiomas recurred.

Objectives: To evaluate mice as experimental animal for Chlamydia pneumoniae, a common cause of acute respiratory infections in human.　Methods: Intranasal inoculation of Icr mice with C. Pneumoniae induced a prolonged course of lung infection, as demonstrated by persistence of lung pathology(60 days).　Results: Icr mice were susceptible to C. pneumoniae. Lung pathology was characterized by patchy interstitial pneumonitis with predominately neutrophil leukocyte infiltration in the early(7 days) and lymphocytes infiltration in the later stages(14 days later) of infection.　Conclusions:Icr mice were susceptible to C. pneumoniae and the mouse model is useful for the investigation of the pathogenesis of C. pneumoniae infection.

Objeetive To investigate the relationship among social avoidance and acceptance of disability in burn patients.Methods Totally 127 patients were investigated using Social Avoidance and Distress Scale (SADS),Acceptance of Disability Scale (AODS),demographic information questionnaire and disease related information sheet.Results The points of burn patients' SADS was (21.38±6.75),One-way ANOVA showed that gender,marital status,special parts burns,amputation affected social avoidance significantly.The total score and factors scores of acceptance of disability were all negatively related to the score of social avoidance.Regression analysis showed that acceptance of disability,head burns,gender were the independent risk factors of depression.Conclusions Acceptance of disability were negatively related to social avoidance; Nurse can help them accept the reality of disability,develop the confidence to partici-pate in social communication,and improve the social adaptability.

Objective In order to play the role of pathological network management system better in pathological examination, this study explore the present status of new pathology network management system, give an objective evaluation for the operation condition, reveal the effectiveness and the existing problems of this system, and provide reference for its development and improvement.Methods The software of pathological network management system was applied to the pathological specimen reception, patient information and examination status query, pathological diagnosis and technology process, as well as the paraffin block archive, statistical analysis, data recording, and so on.At last, we recorded all the information and made a classification and arrangement.Results Pathological network management system was running normally through the whole process of pathologic examination, including specimen receiving, all examinations, print of pathological applications and spontaneous print of pathological reports in ward, which really achieve one-stop services.But the system has unstable phenomenon occasionally.Conclusion Pathological network management system links each examination process closely, which can improve the work efficiency, and provide scientific basis for pathology quality control.

Objective When pathologists from hospitals at various levels encounters pathological sections diffcult to make clear diagosis, it is necessary to invite pathologists from higher hosiptals or special hospitals for pathologic consultation.In the study, we compared the pathological diagnosis of cases sent to other hospitals for pathological consultation with the original diagnostic result to analyze the differences by the evaluation on the impact of these differences on the treatment and prognosis of these patients, which would provide an effective evidence for the quality control of pathological diagnosis. Methods Cases initially diagnosed at the de-partment of Nanjing General Hospital and later sent to other hospitals for pathological consultation from 2010 to 2014 were collected. All the diagnostic results were examined by at least 3 senior pathologists to find exact diffrences between consultation results and origi-nal diagnostic results. Results Among 2055 cases, it was found that there were 1813 cases (88.2%) without diagnostic discrepan-cy, while 218 cases (10.6%) with minor diagnostic discrepancy and 24 cases (1.2%) with completely distinct diagnostic results. Conclusion The diagnostic results of the vast majority of consultation cases are in accordance with the original results, despite of di-agnostic discrepancies in some cases due to the complexity of disease. Expert consultation has reference for the pathological diagnosis of complicated cases, which also plays a potent supervisory role on the quality control of original pathologic results.

Objective To study the effects on berberine hydrochloride on the mitochondria membrane potential and free intracellular calcium of HaCaT cells, and elucidate the mechanism of action of berberine on keratinocytes. Methods Rhodamine-123 fluorescence (very sensitive to mitochondria membrane potential) and Fluo-3/AM fluorescence (suitabe to detect free intracellular calcium in single HaCaT cell) were measured by laser scanning confocal technique. Results Fluo-3/AM fluorescence intensity of HaCaT cells was persistently increased after treating with berberine at concentrations of 5 ? 10-5M, 2.5 ? 10-5M and 1.25 ? 10-5M, and significant differences were observed as compared with the PBS control. The intensity of rhodamine-123 fluorescence in HaCaT cells was decreased immediately when exposed to berberine, with significant difference from that of the PBS control. Conclusions It is suggested that berberine could increase free intracellular calcium and decrease mitochondria membrane potential of HaCaT cells, induce overload of intracellular calcium, influence energy metabolism, and then inhibit the proliferation of keratinocytes.