Objective To explore the diagnostic value of telomerase activity (TA) and hTERT expression in pleural effusions. Methods According to the final diagnosis, 51 patients with pleural effusion were classified into two groups: malignant group (n=27) and benign group (n=24). Telomerase activity in pleural effusion was measured by TRAP-ELISA. Then the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of telomerase activity for diagnosing pleural effusion were calculated. hTERT in pleural effusion was detected by western blotting analysis. Results TA assay showed that 24 of 27 cases of malignant pleural effusion were found to possess telomerase activity, while 1 of 24 cases of benign one had telomerase activity. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of TA assay for diagnosing pleural effusion were 88.9%, 95.8%, 96%, 88.5%, and 92.2%, respectively. hTERT test showed that 8 of 27 cases of malignant pleural effusion had hTERT expression, while none of 12 cases of benign one had hTERT expression. There was significant difference in diagnosing pleural effusion between two methods (? 2=19.64,P=9.4?10 -5 ). Conclusion Tolomerase activity assay may act as a routine diagnostic method for malignant pleural effusions, while hTERT test may serve as complementary one.

Objective:To investigate the inhibitory effect of antisense human telomerase RNA(hTR) gene on implanted hepatocellular carcinoma in nude mice.Methods: HepG2 cells were subcutaneously inoculated into BALB/c nude mice at the axilla to establish implanted hepatocellular carcinoma model.The retrovirus plasmid containing antisense telomerse RNA(PLXSN-hTR-BamHⅠ) was injected into the tumor(0.2 ml every time,5 times).Retrovirus plasmid containing sense telomerase RNA(PLXSN-hTR-EcoRⅠ) and normal saline were inoculated as control groups.Tumor volume was determined and the inhibitory rate was calculated.Tumor necrosis was observed by histological analysis and cell apoptosis was analyzed by terminal transferase dUTP nick end labeling(TUNEL).Results: Tumor growth in antisense hTR group was significantly inhibited compared with the two control groups.The tumor inhibitory rate(26.78%) of antisense hTR group was significantly higher than that of sense hTR group(1.93%,P

OBJECTIVETo explore the venous angio-architecture of deep inferior epigastric artery perforator ( DIEP) flap with computed tomography angiography( CTA) and the application of venous angio-architecture in flap thinning of DIEP flap.

METHODSFrom February 2012 to August 2014, preoperative CTA examination of venous system of DIEP flap was performed in 11 patients to study the course, the branch and the anastomosis of the deep inferior epigastric veins, the superficial inferior epigastric veins and the perforator veins. The flap thinning of DIEP flap was based on the finding of the abdominal venous system.

RESULTSThe CTA images were obtained including 22 deep inferior epigastric veins, 22 superficial inferior epigastric veins and 22 perforator veins. The distance between the superficial inferior epigastric vein and abdominal midline was 3.30 cm to 5.77 cm in the zone 3 cm above umbilicus and in the zone 15 cm below umbilicus. The superficial inferior epigastric vein was 0.29 to 0.39 in depth ratio. The superficial inferior epigastric veins were the main drainage vessels of superficial abdominal wall according to the CTA images. They were anastomosed with the deep inferior epigastric veins through perforator veins. The 11 DIEP flaps were thinned based on the structure of the superficial inferior epigastric veins and all flaps had no drainage disturbance.

CONCLUSIONSPreoperative CTA images can display the venous angio-architecture of deep inferior epigastric artery perforator flap. The superficial inferior epigastric vein can be utilized as reference architecture to guide flap thinning of DIEP flap.

Abdomen ; blood supply ; Abdominal Wall ; blood supply ; Angiography ; methods ; Epigastric Arteries ; anatomy & histology ; diagnostic imaging ; Humans ; Perforator Flap ; blood supply ; Phlebography ; methods ; Tomography, X-Ray Computed ; Veins ; anatomy & histology

BACKGROUND: Some studies have showed that after indirect co-culture, bone marrow mesenchymal stem cells can differentiate into myocardial cells and hepatocypte-like cells. OBJECTIVE: To investigate the possibility of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) to differentiate into hepatocytes by co-culture with human hepatocyte line LO2 in vitro.METHODS: Full-term umbilical cord blood samples were obtained sterilely. The UCB-MSCs were isolated by density gradient centrifugation and directly adherence growth, then passaged with trypsin digestion at 80% cell fusion. By utilizing cell culture plate insets with microporous membrane combined with 6-well plate, the LO2-/UCB-MSCs co-culture system was established. UCB-MSCs were plated into the wells of 6-well plate at a density of 1×10~7/L. LO2 cells were plated into the cell culture plate insert at a density of 1×10~5/L. UCB-MSCs were plated in both layers in the control group. Surface markers of adhered cells were detected by flow cytometry. Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. mRNA expression was detected by reverse transcriptase PCR. RESULTS AND CONCLUSION: HUCB MSCs expressed CD44 and CD29 strongly, but CD34 and CD45 were expressed negatively. After 5 days, fusiform-shaped cells were reduced in the co-culture group; while, the time passing by, cells shaped irregular round or polygonal were increased, which were similar to hepatocytes. At 4 weeks after culture, UCB-MSCs were still fusiform-shaped in the control group. At day 5 after culture, alpha fetoprotein mRNA expressed positively, but other expressed negatively in the co-culture group; at day 14 after culture, cytokeratin-19 mRNA and albumin mRNA expressions were observed; moreover, with the time passing by, the expression of albumin mRNA was increased, but the expression of alpha fetoprotein-19 mRNA was decreased. Antigenic expressions in the control group were negative. This suggested that UCB-MSCs could differentiate into hepatocypte-like cells by co-culture with human hepatocyte line LO2 in vitro.

Objective To compraison the of exposure in the endonasal transsphenoidal approach to the sellar between microscope and endoscope. Methods Ten formalin-fixed, silicone-injected adult cadveric heads were studied. A direct endonasal transsphenoidal approach was performed via the right nostril, pushing aside the nasal septum, then reach the sphenoidal sinus. The approach was performed with the operating microscope first, then with the endoscope. For each step (sellar, suprasellar, parasellar and clival), the operative region afforded by direct microscopic view was measured and then compared with that obtained by using the edndoscope. Results It was found that the endoscope provided greater view than microscope in this approach. Although the microscope provides an adequate view of the midline structures and part of the contralateral parasellar areas; under direct endoscopic vision, the lateral extension could be widened by an additional 6.5 mm on the ipsilateral and 4 mm on the contralateral side. At suprasellar region, the microscope provides could expose the posterior part of, optic nerve and optic chiasma; but could not expose the areas anterior and superior the interspace superior the optic chiasma. Compare with the microscope, the endoscope allowed extension of bone removal and dual opening for an additional 4 mm anteriorly at the sagittal axis and an additional 3.5 mm on the ipsilateral and 4 mm on the contralateral side. At the clivus region, the medial surface of the vertical segment of the ICA and the basilar artery could be partially 7 exposed by the microscope. By the endoscope, it could gain an additional 4 mm on the ipsilateral side and 2.5 mm on the contralateral side in width. Because of the anatomical boundaries of the sphenoid sinus, the anatomincal exposure by the microscope same as the endocope at the sagittal axis. Conclusion The endoscope allows for a panoramic view and permits widening of the operative exposure in all directions. The endoscope is more suitable in the the minimal and expanded endonasal transsphenoial approach.

Objective: To explore flap thinning based on the study of the arterial structure and blood perfusion of the deep inferior epigastric artery perforator (DIEP) flap using computed tomography (CT) angiography. Methods: Clinical imaging study: Preoperative CT angiography was performed in 15 patients with DIEP flap reconstruction to investigate the vascular structure of arterial perforator. Cadaveric imaging study: 10 abdominal specimens harvested from fresh cadavers were cannulated with trocar and injected with contrast medium in the deep inferior epigastric artery perforator. During the perfusion of the contrast medium in the flap, the flap was scanned by three-dimensional CT. The CT data was then sent to CT workstation and the images were processed and reformatted to study the vascular structure of arterial perforators and the blood perfusion. Results: 75 artery perforators in clinical study and 40 artery perforators in cadaveric study were chosen and analyzed. The major deep inferior epigastric artery perforators run directly across the deep layer of adipose tissue without bifurcating beneath the Scarpa′s fascia. Above the Scarpa′s fascia, the artery perforators bifurcate and ultimately terminate in the subdermal vascular plexus. Blood perfusion mode: The subdermal vascular plexus served as the only pathway for blood perfusion between perforasomes. There are two different pathways for blood perfusion in the perforasome: the subdermal plexus and the existing vascular structure of perforator. Conclusions Based on the vascular structure of arterial perforator and blood perfusion of the DIEP flap, thinning of the DIEP flap under the Scarpa′s fascia is safe while thinning above the Scarpa′s fascia should performed according to the blood supply zone of the DIEP flap.

OBJECTIVETo estimate the broad-sense heritability of the production of Glycyrrhiza uralensis and the content of glycyrrhizin as well as the genetic relationship of various growth indexes and biomass indexes, and provide the scientific basis for establishment of high quality licorice cultivate technology system.

METHODThe randomized method was used to assign the provenance trial, the content of glycyrrhizin was determined by HPLC, and the method of classic genetics was applied to estimate the broad-sense heritability and genetic correlation coefficient.

RESULT AND CONCLUSIONThe content of glycyrrhizin is influenced by the growth environment and gene, but the growth environment is the dominant factor. The estimated result of single sites about broad-sense heritability (h2) showed that the production of G. uralensis (W(u)) and the content of glycyrrhizin was controlled by gene which the broad-sense heritability was 0.663 2, 0.751 1 respectively, they had some potential on genetic modification. The results of genetic analysis correlation showed that the plant height and the stem diameter was positive (P < 0.01) correlated significantly with the production (W(u)) either on phenotype or on genetic, it suggests that the plant height and the stem diameter could be the index above ground to assessment the production of the G. uralensis. The content of glycyrrhizin had a positive correlation with the number of lateral root (P < 0.05), but it had a negative correlation with the plant height, stem diameter, diameter of root top (D(r)), the total biomass (W(t)) and the biomass underground (W(u)) on inheritance. It is suggested that it was difficult to achieve both high content and high yield simultaneously in the genetic improvement, so we should have a deeply thought about the specific improvement target when making the reformed scheme.

Biomass ; Glycyrrhiza uralensis ; chemistry ; genetics ; growth & development ; Glycyrrhizic Acid ; analysis ; metabolism

Mutations of the CMT genes develop a variety of distinct phenotypes. Cx32 gene mutations cause the X-linked form of CMT disease, and mutations in EGR2 are associated with CMT type 1, DSS, and congenital hypomyelination neuropathy. Her parents, grandmother and sister did not show the V136A mutation in Cx32. We report the first CMT patient with EGR2 and Cx32 mutations.
Charcot-Marie-Tooth Disease ; Humans ; Parents ; Phenotype ; Siblings

OBJECTIVEIn order to understand the glycyrrhizin content range in the wild and cultivated Glycyrrhiza uralensis in China and to find the related influencing factors of glycyrrhizin content.

METHODThe glycyrrhizin content of 165 wild and 1 013 cultivated G. uralensis samples from 37 countries in 9 provinces was determined by HPLC, and the effects of the producing region, medicinal parts, cultivation years, soil type and texture on the glycyrrhizin content were analyzed.

RESULT AND CONCLUSIONThe average glycyrrhizin content was (4.43 +/- 1.32)% in the wild G. uralensis population, and (1.51 +/- 0.49)% in the cultivated and the glycyrrhizin content in the cultivated was less than the minimum sandards in the Chinese Pharmacopoeia. The glycyrrhizin content was significant different in the wild and cultivated G. uralensis in different producing regions, respectively. The glycyrrhizin content in roots and rhizome of the wild G. uralensis had no significant difference, it had no significant difference in the cultivated G. uralensis from 1 to 4 years and it increased rapidly after 5 years, and the effects of the soil types and texture on it were significant.

China ; Glycyrrhiza uralensis ; chemistry ; growth & development ; Glycyrrhizic Acid ; analysis

Objective: To establish the method of simultaneous determination of contents of honokiol, magnolol and piperitylmagnolol in Cortex Magnoliae Officinalis from different regions, which could provide evidence for the quality control evaluation of Cortex Magnoliae Officinalis. Methods: The quantitative analysis was performed with a column of Waters Acquity UPLC BEH-Cl8 (2.1 mm×50 mm, 1.7 μm) by UPLC-PDA, and eluted with a mobile phase of 0.2％formic acid solution (A)-methanol (B) in a gradient mode (0-2 min, 65％-65％ B; 2-3 min, 65％-75％ B; 3-5 min, 75％-84％ B; 5-9 min, 84％-90％ B) under a flow rate of 0.5 mL·min-1. The column temperature was set at 35℃, and the detection wavelength was 294 nm and the injection volume was 1 μL. Results: The 3 lignan components were completely separated and could be separated well from other components. The honokiol, magnolol and piperitylmagnolol showed a good linear relationship with chromatographic peak area in range of 20.3-406.0, 15.2-304.0, 5.6-112.0 ng, respectively.The average recoveries were 91.75％, 93.86％, 95.15％ with the RSDs were 1.75％, 1.88％, 1.91％ (n = 9), respectively.Conclusion: Contents of the 3 constituents from different place were significantly different and this method is simple and effective, which can be used to quality control of Cortex Magnoliae Officinalis.