BACKGROUND: Differentiation of bone marrow stromal cells (BMSCs) into neurons requires two processes: orientation and differentiation. Orientation and differentiation are results from different gene expression in cells with the same gene bank. Gene expression requires a certain condition. Changes in extracellular matrix can induce changes in cell morphology and gene expression manner. OBJECTIVE: To explore the possibility of BMSC differentiation into neuron-like cells under tissue extract from rat damaged brain. METHODS: The fifth passage of green fluorescent protein (GFP)-transfected BMSCs was induced to differentiate in brain tissue extract from ischemia/reperfusion rats or normal rats. A blank control was set. Cell morphology change was observed under the phase contrast microscope, and then evaluated using immunohistochemical staining. RESULTS AND CONCLUSION: Primarily cultured BMSCs were purified and amplified, and then showed even spindle shape. The third passage of BMSCs was positive for CD44 and CD106, but negative for CD34. Under the fluorescence microscope, BMSCs showed fluorescence expression, but the strength was weak 24 hours following GFP transfection. Numerous cells presented significant green fluorescence 48 hours later. Following adding brain tissue extract from ischemia/reperfusion rats. Induced cells presented neuron-like feature, but neuron specific enolase specific antibody presented positive expression. Compared with the blank control group, the differentiation rate of BMSCs was significantly increased in the ischemia/reperfusion group and normal group (P < 0.05). The increased range was significantly greater in the ischemia/reperfusion group than the normal group (P < 0.05). These results indicated that brain tissue extract from ischemia/reperfusion rats can successfully induce the differentiation of BMSCs into neuron-like cells.

Objective: To observe the clinical effect of tuina in rehabilitation following total hip replacement. Methods: 60 cases were randomly allocated into a treatment group and control group, 30 cases in each group. Cases in the treatment were treated with combined mina and rehabilitation, while cases in the control group were treated with rehabilitation alone. All treatment retained for two weeks. The results were observed 7 d, 2 weeks and 6 weeks following the total hip replacement using Harris scale and Hamilton Anxiety Rating Scale (HAMA).Results: For Harris scale, there were significant intra-group differences in different time frames (P＜0.05); there were no statistical differences between the two groups 1 d, 7 d and 6 weeks following the total hip replacement (P＞0.05); and there were statistical differences between the two groups 2 weeks following the total hip replacement. For HAMA scale, there were significant intra-group differences in different time frames (P＜0.05); there were no significant differences between the two groups 1 d following the total hip replacement; and there were substantial differences between the two groups 7 d, 2 weeks and 6 weeks following the total hip replacement. Conclusion: In a given unit time, combined mina and rehabilitation is superior to rehabilitation alone in improving the patients' post-operative pain, articular range of motion and anxiety.

OBJECTIVE To understand the synaptic remodel in inner ear after hearing injury by investigating growth associated protein 43(GAP-43) expression in the inner ear hair cells after exposed Kangmin mice to neural injury noise stimulation to induce permanent threshold shift(PTS) or temporary threshold shift(TTS).METHODS To compare the expressions of GAP-43 of NMDAR signal pathway during neural injury stimulation by immunocytochemistry.RESULTS The sustained GAP-43 increase occurs after 7d and 14d of PTS group,and there is not remarkable difference between them.Compared with normal control,the change is not significant in TTS group.CONCLUSION The increase of GAP-43 in spiral ganglion after 7d trauma reveals the synaptic remodel in inner ear.

Dunhuang medicine is a branch of Dunhuang studies, including the characteristics of Dunhuang ancient medical prescriptions and treatment technique.Dunhuang Dabupi Decoction has good efficacy in nourishing vitality and yin and promoting the production of body fluid. Professor WANG Dao-kun used the prescription to treat gastric stuffiness, gaining significant curative effect. This article summaried cases of using this prescription, to show the advantages of Dunhuang ancient medical prescription Dabupi Decoction for present use.

Objective To systematically investigate the molecular epidemiological profiles of human papillomavirus (HPV) in patients with condyloma acuminata(CA). Methods Two hundred and one samples of HPV DNA isolated from CA were PCR amplified by the PGMY09/11 primer system. The PCR products were simultaneously hybridized to 37 specific HPV probes immobilized on a nylon strip and then genotyped. All DNA templates were further PCR amplified using HPV 6 and 11 type specific primers for verification. Results All samples were HPV DNA positive consisting of totally 31 genotypes, the types of which were type 11(53.7%, 108/201), 6(43.8%, 88/201), 16(6.5%, 13/201), 52(6.0%, 12/201), 33(5.5%, 11/201), cp6108 (5.5%, 11/201) and 42 (5.0%, 10/201). The samples infected with a single and mixed types of HPV accounted for 60.2% (121/201) and 39.8% (80/201) respectively. Consistent results were found with the detection of HPV6 and 11 between hybridization assay and type-specific PCR. Conclusions At least 31 HPV genotypes are associated with CA. HPV 11 predominates while 68, 40, 54, 67, 73, 82, 35, 64 and 83 are rare in CA. Type cp6108 is detected in CA for the first time with a high prevalence. HPV26, 69, 70, 71,72 and IS39 might be not associated with CA. CA infected with a single and mixed HPV types accounts for 60.2% and 39.8%, respectively.

Objective:To compare the differences in the delineation of the gross tumor volume (GTV) and lymph nodes of nasopharyngeal carcinoma (NPC) patients using computerized tomography (CT), magnetic resonance imaging (MRI), and 18F-fluorodeoxyglucose positron emission tomography/computed tomography ( 18F-FDG PET/CT), and to investigate the optimal standard uptake value (SUV; relative to the MRI-based delineation) for the automatic delineation of GTV using PET. Methods:A total of 53 NPC patients proposing to receive radiotherapy were selected for this study. The CT, MRI, and PET images of each patient were obtained before radiotherapy. Then GTV and positive lymph nodes were delineated on these three types of images. They were individually named GTV MRI, GTV CT, GTV PET2.5 (SUV=2.5), Lymph MRI, Lymph CT, and Lymph PET2.5 and compared. The GTV ∩2.5 (overlapped GTV) was obtained through the alignment of MRI and PET/CT images. Meanwhile, GTV was delineated on PET images using thresholds of SUV=4.0, 4.5, 5.0, and 5.6, obtaining GTV PET4.0, GTV PET4.5, GTV PET5.0, and GTV PET5.6. Then their volume and Dice similarity coefficients (DSCs) were compared. Results:Compared to GTV MRI, GTV CT decreased by 1.73% ( P>0.05) and GTV PET2.5 increased by 21.34% ( t=-3.52, P < 0.05) in the three types of images. The volume of Lymph PET2.5 was 1.61 and 1.87 times the volume of Lymph MRI and Lymph CT, respectively ( t=-4.12, -5.18; P< 0.05). The volume of high-SUV lymph nodes was 4.07 times the volume of lymph nodes with low SUVs or SUV=0 ( t=5.50, P< 0.05) on PET images. The DSC between GTV PET4.0and GTV MRI was 0.78 ± 0.27, which was lower than that between GTV PET2.5 and GTV MRI (0.84 ± 0.18). However, GTV PET4.0 approximated to GTV ∩2.5 ( P>0.05). Conclusions:Compared to CT and 18F-FDG PET/CT, MRI shows more accurate boundaries of GTV and lymph nodes. When 18F-FDG PET/CT was adopted to automatically delineate GTV, the GTV delineated using SUV=4.0 was closer to GTV MRI.

OBJECTIVETo set up a stable primary culture system of Leydig cells with higher purity.

METHODSWe separated Leydig cells from other testicular cells, such as Sertoli and germ cells, by enzymatic digestion in combination with Percoll density gradient centrifugation and identified Leydig cells by 3beta-HSD staining.

RESULTSThe purity achieved by this method was above 95% and the total number of Leydig cells obtained from one testicle was about 1 x 10(6). The cytoplasm of Leydig cells was stained in deep blue by 3beta-HSD staining, and these cells possessed testosterone-secreting capability.

CONCLUSIONLeydig cells can be separated by enzymatic digestion combined with Percoll density gradient centrifugation, and 3beta-HSD staining to identify Leydig cells is simple and feasible with high purity.

3-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Cell Culture Techniques ; Cell Separation ; methods ; Cells, Cultured ; Centrifugation, Density Gradient ; methods ; Histocytochemistry ; Leydig Cells ; cytology ; enzymology ; physiology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.

METHODSAfter one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.

RESULTSThe NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).

CONCLUSIONSLXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.

Animals ; Berberine ; therapeutic use ; Diet, High-Fat ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; Hepatocytes ; Lipids ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Objective To evaluate the efficacy of Epipolis laser in situ keratomileusis(Epi-LASIK)and laser in situ keratomileu- sis(LASIK)on high myopia patients.Design Prospective,case-controlled study.Participants 62 patients(123 eyes)with high myopia. Methods 62 patients(123 eyes)underwent Epi-LASIK or LASIK surgery for high myopia:28 patients(56 eyes)underwent Epi-LASIK and 34 patients(67 eyes)underwent LASIK.The differences in postoperative uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),refraction,and root mean square(RMS)of high-range wavefront aberration were compared one-week,one-month, three-months and six-months postoperatively.Main Outcome Measures UCVA,BCVA,refraction,and RMS of high-range wavefront aberration.Results There was no serious complication during and after the operation.The recovery of postoperative UCVA after Epi-LASIK was slower than that of LASIK.One week postoperatively,the proportion of UCVA≥0.8 of Epi-LASIK(46.4%)was less than that of LASIK(77.6%)(P=0.0003).No significant differences were found in those of Epi-LASIK(85.7%,94.6%,91.1%)and those of LASIK(92.5%,95.5%,94.0%)one-month,three-months and six-months postoperatively(P=0.590,0.822,0.530).BCVA was same after Epi-LASIK and LASIK.The proportion of mean spherical equivalents within?0.50D for Epi-LASIK(42.9%,51.8%,60.7%,64.3%)had no difference with those for LASIK(53.7%,59.7%,71.6%,73.1%)one-week,one-month,three-months and six-months postoperatively (P=0.230,0.378,0.200,0.290).The postoperative RMS increased after both surgeries,especially after LASIK.At postoperative one-month,three-months and six-months RMS of Epi-LASIK(1.51?0.77)?m,(1.32?0.76)?m,(1.18?0.71)?m were much higher than the (0.87?0.27)?m preoperative ones(P=0.016,0.019,0.026).At postoperative one-month,three-months and six-months RMS of LASIK (2.41?0.81)?m,(2.17?0.63)?m,(1.89?0.87)?m were also much higher than the preoperative(0.91?0.22)?m(P=0.011,0.008,0.006). There were significant differences between the RMS of Epi-LASIK and LASIK one-month,three-months and six-months postoperatively (P=0.039,0.035,0.033).The I grade haze was found in two eyes after Epi-LASIK.Conclusion Epi-LASIK has better visual quality re- sult than LASIK on correcting high myopia.(Ophthalmol CHN,2007,16:336-339)

Objective:To evaluate the cumulative dose of the target volume and organs at risk (OARs) in intensity-modulated radiation therapy (IMRT) for large volume non-small cell lung cancer (NSCLC) based on rigid and deformation registration methods. The dosimetric changes between the initial and second treatment plans were compared.Methods:Thirty patients treated with IMRT for large volume NSCLC with twice 4DCT scans acquired before radiotherapy and after 20 fractions of radiotherapy were recruited. The initial treatment plan (Plan 1) based on the average density projection CT (CT 1-avg) of the first 4DCT images and the second treatment plan (Plan 2) based on the average density projection CT (CT 2-avg) of the second 4DCT images were calculated. Then, the dose distributions of Plan 1 and Plan 2 were accumulated based on rigid and deformation registration methods to obtain Planrig and Plandef, respectively. Finally, the volume changes of gross tumor volume (GTV) and OARs between two CT scans were compared. The dose-volume parameters between Plan 1 and other plans (including Plan 2, Planrig and Plandef) were also statistically compared. Results:Compared with the initial CT scan, the mean volume of GTV and heart on the second CT was decreased by 44.2% and 5.5%, respectively, while the mean volume of ipsilateral lung, contralateral lung and total lung was increased by 5.2%, 6.2% and 5.8%, respectively (all P<0.05). Compared with Plan 1, the D 95%, D 98% and V 100% of target volume IGTV (GTV fusion of 10 4DCT phases) and PTV in Plan 2 did not significantly change (all P>0.05), and those in Planrig and Plandef were decreased (all P<0.05). The dose-volume parameters of spinal-cord, heart, ipsilateral lung and total lung in Plan 2, Planrig and Plandef were significantly lower than those in Plan 1(all P<0.05). Among them, the V 30Gy and D mean of heart were decreased by 27.3%, 16.5%, 15.3% and 15.2%, 6.6%, 5.6%, respectively. The V 20Gy and D mean of total lung were decreased by 15.6%, 4.5%, 3.7% and 15.7%, 6.2%, 5.1%, respectively. Some dose-volume parameters (including D 95% and D 98% of target volume, V 40Gy of heart, V 20Gy and D mean of the ipsilateral lung and the total lung) of Plandef were higher than those in Planrig (all P<0.05). The Dice similarity coefficients (DSCs) of OARs after deformation registration were significantly higher than those after rigid registration ( P<0.05). Conclusions:The dose-volume parameters of OARs significantly differ between Plan 1 and Plan 2. Hence, all these parameters have a large degree of deviation in predicting radiation-induced injury of OARs. Nevertheless, the dose-volume parameters obtained by deformation registration can enhance the prediction accuracy.