Objective To study the clinicopathological characteristies,differential diagnosis,etiological factor and treatment of anorectal malignant melanoma(AMM). Methods Clinical pathological characteristics and immunohistochemical findings were observed in 13 cases of anorectal malignant melanoma,with review of related literature.Results Average age was 55.8 years. And often occurred in the middle and old. Under microscope, the tumor was complicated constructionally, and the tumor cells were variform. Usually in tightly packed nests, dispersed sheet and trabecular, melanin pigment could be observed in the majority of AMM, but sometimes ameLanotic pigment was present. Immunohistochemiscal the tumor cells were positive for HMB 45, S-100 and vimetin. Conclnsion AMM is a high malignant tumor with very poor prognosis. So,it should be diagnosed and treated early. HMB 45 and S-100 are sensitive tumor markers for AMM.

Objective To investigate the correlations of the expressions of CXCR4 and vascular endothelial growth factor (VEGF) in esophageal cancer and their associations with patients' clinicopathological features.Methods The paraffin-embedded samples of 127 patients with esophageal cancer treated by thoracic surgery in Zhongshan Hospital in 2005 were collected. The CXCR4 and VEGF expressions in esophageal cancer tissues were detected with immunohistochemical technique. The immunostaining was evaluated according to the staining area and intensity. Results The overall expression rate was 88.2% and 79.5%, respectively. CXCR4 expression was significantly associated with tumor grade, size, tumor depth, regional lymph node metastasis and TNM stage (P<0.05);VEGF expression was significantly associated with tumor depth, TNM stage and tumor grade.In addition, there were significant correlations between CXCR4 and VEGF (P<0.01). Conclusions There was significant association between CXCR4, VEGF expressions and clinicopathological features of esophageal cancer. There might be some potential correlation between CXCR4 and VEGF expression in esophageal cancer.

Chitosan, deacetylated derivative of chitin, is a kind of natural polysaccharide polymer. It has advantages of rich source, good biocompatibility and biodegradation. Chitosan can be processed into porous scaffolds used for cell transplantation and tissue regeneration in bone tissue engineering, control-released carrier of growth factors and delivery vector of exogenous genes. Meanwhile, it can also be processed into injectable scaffolds used in bone tissue engineering in the form of microspheres or hydrogel. Chitosan and its derivatives will have broad application prospects in the research field of bone tissue engineering. However, chitosan composite scaffold has poor mechanical function, difficult accuracy control of In vivo degradation, and low efficiency for genetic carrier, chitosan research stays in in vitro tests and in vivo animal experiments. With the development of materials science and life science, chitosan will be widely used for clinical application of bone tissue defect.

Objectivo To analyze urine organic acids in the urine using gas chromatography-mass spectrometry(GC/MS)for diagnosis of inherited metabolic diseases,especially for organic acids metabolic disorders.Methods 195 clinical urine samples from the patients with suspected organic acids metabolic disorders and 5 normal urine from adults were collected.After mixing some urine with intemal standards according to the concentration creatinine and adding hydroxylamine hydrochloride to mixture,the organic acids with hydroxyl group were oximated to the ketobodies.Organic acids were extracted twice with ethyl acetate and ethyl ether and derivatized with BSTFA-TMCS.An the organic acids were determined with Agilent GC/MS 6890/5973i with scan model.the mass-to-charge ratio range is 50-550 m/z,all data were nalyzed with Agilent GCMSD ChemStationG1701DA.We also investigated the linearity, accurate,precision.recovery and Carry-over by determining the internal standards in normal samples and positive organic acids in spiked control samples.Results More than one hundred kinds of organic acids in urine samples can be analyzed with this method.According to the two internal standards in normal urine samples,the minimal detection limit MMA and 2.PA was 2.5-2.8 μmol/L.Intra-and interassay coefficient of variation for MMA and 2-PA are both less than 10%.Pre-processing Interassay coefficient by sequential preparations of the same sample was 14%.The recoveries of the spiked samples were 95%-105%.Carryover analysis was less than 1%.All the parameters meet the requirement for clinical diagnosis.12 samples demonstrated positive including 6 cases of methylmalonic acidemia,1 case of propionic acidemia,3 cases of tyrosinemia Ⅰ,1 case of maple syrup urine disease and 1 cases of ketosis.Conclusions The method for the determination of organic acids in urine by GC/MS has been successfully established.It can be used for clinical screening and diagnosis for inherited genetic metabolic diseases.

Objective:To investigate the in vitro effects of platelet-rich plasma(PRP) on human periodontal ligament fibroblasts(PDLFs). Methods: Various concentrations of PRP (10, 50, 100, 200, 300, 500 ml/L) were applied to primary cultures of human PDLFs. MTT assays were utilized to assess cell proliferation ability. Migration was determined by assessing the cell response to a concentration gradient with Transwell chamber. Differentiation was assessed using alkaline phosphatase (ALP) kit. Results: A beneficial effect on proliferation was observed, especially in response to 200 ml/L PRP.PRP had stimulatory effects on the migration of human PDLFs. PRP facilitated differentiation of PDLFs. Conclusion: PRP can exert a positive effect on human PDLFs,but this effect is concentration specific, while higher concentrations is not necessary to result in optimal outcomes.

Objective To package the recombinant lentivirus containing HIV-1 Vpr gene and detect the effect of Vpr protein expression on the latent infection and lyric replication of KSHV.Methods The fragment of Vpr gene from expression plasmid pCI-neo-Vpr was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen,then the recombinant plasmid pHAGE-Vpr,package vector psPAX2 and envelope vector pMD2.G were cotransfected into 293T cells.GFP expression was observed by fluorescent microscopy.Culture media of 293T cells were harvested and filtered through 0.45 μm filter.After 293T cells were infected by a series of diluted lentivirus,the virus titer was checked by observing GFP expression.Vpr mRNA transcripts in 293T cells were detected by RT-PCR.Then BCBL-1 cells were infected by the recombinant lentivirus with 1 MOI,GFP expression was observed by fluorescent microscopy,and the mRNA transcripts and protein expression of Vpr in BCBL-1 cells were detected by RT-PCR and Western blot.Meanwhile,the mRNA transcripts and protein expression of KSHV lytic cycle gene Rta were detected by RT-PCR and Western blot,respectively.Results The recombinant lentivirus carrying HIV-1 Vpr was packaged successfully with the virus titer of 4 × 107 TU/ml.After infected with lentivirus,BCBL-1 cells could express GFP,and the exact band of Vpr was detectable by RT-PCR and Western blot.Moreover,the expression of KSHV Rta mRNA and protein were downregulated by Vpr protein.Conclusion Overexpression of HIV-1 Vpr mediated by the recombinant lentivirus could inhibit KSHV lytic replication and enhance KSHV latent infection.

Streptozotocin-induced diabetic rats were treated with GSH for 12 weeks.The results showed that GSH significantly improved the expressions of NF-KB and inducible nitrie oxide synthase and ameliorated the myocardial tissue injury.

Cortex Periplocae Radicis (CPR) is the dried root barks of Periploca spium Bge. It has been used in China for treat­ing rheumatoid arthritis and strengthening the bone and the musculature for thousands of years. Recently, an increasing number of bio­activities of CPR have been recognized, including tumor suppression and anti-chronic heart failure function. However, long-term use or large doses of CPR may produce adverse reactions, which constrains its applications in clinical therapy. Therefore, it is critical to know the chemical components of CPR in order to understand the toxicity mechanism. Nearly a hundred chemical compositions have been found, however, various classes, obfuscated names of different compounds, and disaccord between chemical structure and name were major obstacles to studying pharmacodynamics and toxicity of CPR. In this paper, 94 chemical components of CPR are classified and reviewed, which is valuable for the comprehensive understanding of its biological functions and safe clinical use.

OBJECTIVE Diabetes-induced endothelial cell (EC) dysfunction and neovasculariza-tion impairment constitute vascular complications with limited treatment regimens.Transcription factor FOXO1 is a key angiogenic regulator and plays a pathologic role in progression of diabetes.The pres-ent study was designed to determine the involvement of FOXO1 in impaired EC function and post-isch-emic neovascularization in diabetes and investigate underlying mechanisms.RESULTS We found that FOXO1-selective inhibitor AS1842856 improved blood flow recovery and capillary density in ischemic hindlimb,and rescued the delay of wound closure with a concomitant augmentation of mean perfusion rate in diabetic mice. In vitro,treatment with AS1842856 or FOXO1 siRNA abrogated high glucose-in-duced apoptosis and ameliorated capillary tube formation in human umbilical vein endothelial cells(HU-VECs). FOXO1 inhibition relieved alterations in mitochondrial networks and significantly suppressed the over production of mitochondrial reactive oxygen species(mtROS)induced by high glucose in ECs. Expression of dynamin-relatedprotein-1 (Drp1) and phosphorylation at Ser616, a protein required for mitochondrial fission, were enhanced by hyperglycemia, which could be neutralized by FOXO1 inhibition. Moreover, the transcription of Rho-associated coiled-coil containing protein kinase 1 (ROCK1), which phosphorylates Drp1 at Ser616, was shown by luciferase assay to be directly regulated by FOXO1. CONCLUSION These findings suggested that FOXO1 is critical to preserve mitochondrial quantity and func-tion in ECs,and FOXO1 may serve as a therapeutic target for microvascular complications of diabetes.

Objective To study the therapeutic effect of continuous renal replacement therapy (CRRT) in renal failure after liver transplantation.Methods Renal function in 82 patients who underwent CRRT in the perioperative period of liver transplantation were retrospectively analyzed.Results There were significant differences in ALT,TB,BUN and Cr before and after the treatment (P ＜ 0.05).The differences were significant in glutamic-pyruvic transaminase (ALT),creatinine phosphate kinase (CPK) and C reactive protein (CPR) before and after the treatment (P ＜0.05).There were significant improvements in K+,Na+,Cl-,HCO3-and CVP before and after the treatment (P ＜ 0.05),while the differences were not significant in other biochemical parameters (P ＞ 0.05).This research also looked at the effect of timing of CRRT on renal function recovery.Based on the RIFLE classification of AKI,the ratio of renal function recovery in RIFLE-Ⅰ was significantly higher than RIFLE-F (P ＜ 0.05).Conclusion CRRT treatment significantly improved the prognosis of patients with acute renal failure after liver transplantation.