Objective To investigate the inhibitory effects of traditional Chinese medicine (TCM) on hypermelanosis and provide experimental evidence for treating skin pigmentary disorders. Methods Five TCMs with strong inhibitory effects on tyrosinase activity were tested. The changes of the number and morphology of melanocytes induced by UVB were observed in the experimental hypermelanosis model (brownish guinea pig). Results Decreased melanocytes and melanin granules were found with the treatment of Poria cocos, Polyporus umbellatus, Cornus officinalis Sieb. et Zucc., Atractylodes macrocephala Koidz., Astoagalus complanatus R. Br. ex Bge. Conclusion There are inhibitory effects on hypermelanosis induced by UVB with the treatment of Poria cocs, Polyporus umbellatus, Cornus officinalis Sieb.et Zucc., Atractylodes macrocephala Koidz., Astoagalus complanatus R. Br.ex Bge.

Twelve specimens of genital warts were excised from 12 patients and divided into three parts. One part was untreated, the second and the third part were treated with CO_2 laser and microwave respectively. HPV DNA was amplified and detected in 100% of untreated specimens (HPV16 and HPV11 in six patients each), in 83% and 50% of specimens treated with CO_2 laser and microwave respectively. There was a significant difference in the detection rates between untreated and microwave treated specimens (X~2=4.18, P

Objective To investigate the ultrastructural characteristics of amelanotic melanocytes (AMMCs). Methods Individual hair follicles from normal human scalp were digested with collagenase type V, then washed in phosphate buffer saline. Hair-follicle cell suspensions were prepared by trypsin and cultured in a medium suitable for melanocyte growth. The keratinocytes were removed by differential trypsinization. Geneticin (100?g/mL) was used to eliminate contaminating fibroblasts. After 3 passages the cells were trypsinized, washed in phosphate buffer saline, and finally processed for transmission electron microscopy. Results Under transmission electron microscope, the cultured cells were round or oval-shaped with a single large nucleus and double-layered karyotheca. Abundant euchromosome but sparse heterochromosome was observed within the nucleus. There were various organelles in the cytoplasm, including mitochondria, rough endoplasmic reticulum (RER), ribosomes and abundant melanosomes of nearly uniform size. The electronic density granules distributed in a concentric pattern in most of the melanosomes. Colgi complexes were inconspicuous in the cells. Conclusions Compared to epidermal melanocytes, AMMCs from human hair follicles have different ultrastructural characteristics which implies their functional immaturity. AMMCs may serve as the depot for mature melanocytes.

Aim To study the mechanism of magnolol-induced HeLa cell death.Methods Cell viability was measured by MTT method.Morphological changes were observed by phase contrast microscopy and Hoechst 33258 staining. DNA fragmentation was assayed by agarose gel electrophoresis. Protein level was detected by Western blot analysis.Results Magnolol induced HeLa cell apoptosis and had a weaker cytotoxic effect on HEL 299 cell. The apoptosis of HeLa cells was partially reversed by caspase-3,-8,-9,-10 and caspase family inhibitors. Magnolol has a synergistic apoptotic effect with Fas agonistic antibody CH-11. Treatment of HeLa cells with magnolol for 12 h increased the protein expression ratio of Bax/Bcl-X_L; procaspase-3, ICAD and PARP were cleaved to smaller molecules. p53 and phosphorylation of p53 (ser-15) increased response to magnolol treatment.Conclusion Magnolol induced HeLa cell death via alteration of Bax/Bcl-X_L ratio,activation of caspases and accumulation of p53.

BACKGROUND:Mesenchymal stem cel s are the seed cel s for tendon tissue engineering which can be obtained in large quantities, but how to induce in vitro is a key technology.
OBJECTIVE:To explore the effect of platelet-rich plasma on the proliferation and col agen production of in vitro cultured mesenchymal stem cel s.
METHODS:The rabbit mesenchymal stem cel s were separated ad cultured. The high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group were set to induce the mesenchymal stem cel s, and the blank control group was set as control.
RESULTS AND CONCLUSION:The proliferation of mesenchymal stem cel s in the high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group was high, and in rapid growth with big increase amplitude, and there was no significant difference in the proliferation when compared with the blank control group (P<0.05). The effect was positively correlated with the culture time, and after cultured for a certain time, the effect was in dose-dependent manner, as higher dose platelet-rich plasma had more significant effect on the proliferation of the cel s. The results indicate that platelet-rich plasma can significantly promote the synthesis of col agen type Ⅰ and Ⅲ of mesenchymal stem cel s, the higher the dose, the more significant the effect on the col agen.

Objective To evaluate the correlations of vascular structures,hemodynamic changes and recanalization before receiving carotid endarterectomy ( CEA) in patients with subtotal or complete occlusion of carotid artery using color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) ultrasonography. Methods A total of 107 patients were diagnosed as subtotal ( stenosis rate 95% to 99%) or complete occlusion of carotid artery with DSA and treated with CEA at Beijing Xuanwu Hospital, Capital Medical University from January 2005 to January 2014 were enrolled retrospectively. The mean age of patients was 61 ± 9 years. According to the findings of DSA,they were divided into either a carotid artery subtotal occlusion group (n=63) or a complete occlusion group (n=44). The vascular diameter,the locations of the lesions ( internal carotid artery or common carotid artery) ,the lumen echo characteristics,and whether internal-external artery collateral circulation patent or not at different stages in patients of both groups were documented. Results The lumen diameter of distal segment was significant wider in patients of the complete occlusion group compared with the subtotal occlusion group (4. 1 ± 1. 1 mm vs. 3. 2 ± 0. 8 mm). There was significant difference between the 2 groups (P <0. 01). There was no significant difference between the location of occlusion and the recanalization rate (P=0. 460). The recanalization rate of the lumen homogeneous echo ( hypoecho and echodense) filling patients (94. 1% vs. 86. 7%) was significantly higher than that of the patients of heterogeneity echo. In patients with complete occlusion of internal carotid artery,the recanalization of CEA would increase when the internal-external collateral arteries were patent. For general comparison,the recanalization rate of the subtotal occlusion group was significantly higher than that of the complete occlusion group (P<0. 01). Conclusion The carotid artery diameter normal or broadening ,the homogeneous echo in the occlusive lumen and the internal-external collateral arteries patency are closely associated with the recanalization rate. The preoperative ultrasonography has great value for the assessment of recanalization of carotid artery occlusive disease after CEA.

Objective To investigate the criteria of hemodynamic parameters for diagnosis of intracranial segment vertebral artery stenosis with transcranial color-coded sonography (TCCS ). Methods A total of 622 outpatients or inpatients with suspected posterior circulation ischemia were enrolled retrospectively,from which 216 patients were selected with TCCS,color Doppler flow imaging (CDFI)screen,and digital subtraction angiography (DSA)examination,including 33 patients (15. 3%) had normal intracranial vertebral arteries,the stenosis rates<50% were 45 cases (20. 8%),50%-69%were 44 cases (20. 4%),and 70%-99% were 94 cases (43. 5%). The mean velocity (MV)of intracranial segment,the ratios SPRP (PSV1/PSV2 ),SPRE (EDV1/EDV2 )of the systolic and end diastolic flow velocity between the intracranial segment and the intervertebral space segment were calculated respectively by detecting the intracranial segment of vertebral artery,the intervertebral space segment peak systolic velocity (PSV1 ,PSV2 )and end diastolic velocity (EDV2 ,EDV1 ). The DSA findings were used as the criteria,the area under the receiver operating characteristic (ROC ) curve was calculated and the optimal cut-off points were obtained. Results The optimal cut-off points of TCCS diagnosis of intracranial vertebral artery stenosis were as follows:the parameter standards of stenosis rate <50% were 110 cm/s≤PSV1≤145 cm/s and 65 cm/s≤MV≤85 cm/s,the parameter standards of stenosis rate 50%-69%were 145 cm/s≤PSV1≤190 cm/s and 85 cm/s≤MV≤115 cm/s,and the parameter standards of stenosis rate 70%-99% were PSV1≥190 cm/s and MV≥115 cm/s. Conclusion TCCS may effectively evaluate the hemodynamic changes of intracranial vertebral artery stenosis and provide reference for the ultrasound evaluation criteria of intracranial vertebral artery stenosis.

Objectives Todynamicallyobservethechangesofhemodynamicparametersinpatients with severe stenosis of unilateral middle cerebral artery (MCA)by transcranial Doppler ultrasound (TCD) andtoevaluateandanalyzetherelatedfactorsforinfluencingthestenoticprocess.Methods Atotalof 113 consecutive patients with severe stenosis of unilateral MCA screened by TCD and confirmed by computed tomography angiography (CTA)and digital subtraction angiography (DSA)were enrolled retrospectively. They were divided into either a progressive group (n =43 )or a non-progressive group (n=90)according to the variation of MCA hemodynamic parameters. The effects of age,sex,major risk factors for cerebrovascular disease,clinical symptoms,clinical medication,and drug compliance on the stenotic process were documented and analyzed. Results (1)The comparison of detection rate of the risk factors for cerebrovascular disease:The patients with a history of smoking (72. 1%[n=31])in the progressive group was significantly higher than that (51. 1%[n=46])in the non-progressive group (P=0.022). The period of smoking of the patients in the progressive group were longer than that in the non-progressive group (28 ± 12 years vs. 21 ± 10 years,P=0. 011). (2)Comparison of MCA hemodynamic parameters:The distal pulsatility indexes of MCA stenosis at the first diagnosis in the progressive group were all lower than those in the non-progressive group (0. 66 ± 0. 10 vs. 0. 70 ± 0. 13;t= -2. 096,P=0. 038),and the distal pulsatility indexes of MCA stenosis at the end point in the patients of the progressive group were lower than those in the non-progressive group (0. 61 ± 0. 15 vs. 0. 74 ± 0. 15). There were significant differences (t=-2. 718,P= 0. 008). The peak systolic velocity (PSV)of the progressive MCA stenotic segments at the end point in 10 patients of the progressive group was higher than that in the non-progressive group (299 ± 23 cm/s vs. 244 ± 50 cm/s,t=3. 437;P=0. 001),while PSV of MCA in 33 patients with occlusion in the progressive group were significantly lower than those in the non-progressive group (56 ± 18 cm/s vs. 244 ± 50 cm/s,t= -20. 905;P=0. 000). (3)The regular medication:The patients using statins (atorvastatin calcium)were significantly lower than those of the non-progressive group (2. 3%[n=1] vs. 54. 4%[n=49],χ2 =33. 690;P<0. 01). (4)During the follow up period,the recurrence rates of transient ischemic attack and stroke of the progressive group were significantly higher than those of the non-progressive group (27. 9%[n=12]vs. 6. 7%[n=6],32. 6%[n=14]vs. 2. 2%[n=2];all P<0.01). (5)Multivariate Logistic regression analysis showed that smokers (OR,4. 403,95%CI 1. 094-14.017),cerebrovascular event recurrence (OR,10. 648,95%CI 2. 530 -41. 261),and irregularly taking statins (OR,5. 675,95%CI 1. 631-152. 740)were all closely associated with the progress of severeMCAstenosis.Conclusion EvaluationofthehemodynamicchangesofsevereMCAstenosiswith TCD follow up study can be used as an important basis for clinical assessment of the outcomes. Stop smoking and regularly taking statins may help to delay the progress of MCA stenosis.

Objective To investigate the effects of Xanthoceraside on the learning and memory impairment induced in mire by innacere-broventricular injection of aggregated amyloid β peptide _(1-42)(Aβ_(1-42)). Methods Learning and memory functions in mice were examined us-ing step-through test and water maze test. Biochemical determination of acetylcholinesterase (AchE) and choline acetyl transferase (ChAT) were measured with spectrophotometric melhod. Results Administration of Xanthoceraside reduced number of error and prolonged the laten-cy in step-through test in mice impaired by Aβ_(1-42) (P < 0.05,P< 0.01,respectively). In water maze test,the swimming time decreased in mice treated with Xanthoceraside compared with the model mice impaired by Aβ_(1-42) (P< 0.05,P< 0.01,respectively). The results of bio-chemical determination showed that decrease level of AchE and ChAT in mice impaired by Aβ_(1-42) were significantly ameliorated by Xantho-ceraside administration (P < 0.05 and P < 0.01). Conclusion Xanthoceraside has the effect of improving learning and memory impairment in mice inducel by intracerebroventricular injection of Aβ_(1-42) via enhancing the cholinergic system functions.

Penicillium decumbens T. is an important filamentous fungus for the production of cellulases to effectively degrade lignocellulose for second generation biofuel production. In order to enhance the capability of Penicillium decumbens to produce cellulases, we constructed a creB (a deubiquitinating enzyme encoding gene) deletion cassette, and generated a creB knockout strain with homologous double crossover recombination. This mutation resulted in a detectable decrease of carbon catabolite repression (CCR) effect. The filter paper activity, endoglucanase activity, xylanase activity and exoglucanase activity of the deltacreB strain increased by 1.8, 1.71, 2.06 and 2.04 fold, respectively, when comparing with the parent strain Ku-39. A 2.68 fold increase of extracellular protein concentration was also observed. These results suggest that the deletion of creB results in CCR derepression. These data also suggest that CREB influences cellulase production of Penicillium decumbens. In generation, this study provides information that can be helpful for constructing cellulase hyper-producing strain.
Cellulase ; biosynthesis ; Endopeptidases ; genetics ; metabolism ; Gene Knockout Techniques ; Lignin ; metabolism ; Mutant Proteins ; metabolism ; Penicillium ; enzymology ; genetics ; Recombination, Genetic ; Ubiquitinated Proteins ; genetics ; Ubiquitination