Objective To explore the value of carbon nanoparticles in patients with papillary thyroid carcinoma (PTC) undergoing total thyroidectomy combined with ipsilateral central lymph node dissection.Methods 43 patients with unilateral PTC were retrospectively analyzed.All patients underwent total thyroidectomy combined with central lymph node dissection.Patients were divided into carbon nanoparticles group and the control group according to whether carbon nanoparticles were used in the operation.The operation time,postoperative hospitalization time,the serum calcium level and its rate of change,the parathyroid hormone and its rate of change on the 1 st day after surgery,the number of central lymph node and the transfer rate,and the postoperative complications were compared between the 2 groups.Results There was no statistical difference between the 2 groups in operation time(P ＞ 0.05),while the postoperative hospitalization time of the carbon nanoparticles group was shorter than that of the control group(P ＜ 0.05).The serum calcium and the parathyroid hormone on the 1 st day after surgery in the carbon nanoparticles group was (2.31 ± 0.13) mmol/L and (33.45 ± 14.37) pg/ml respectively,higher than those of the control group (P ＜ 0.05).The low blood calcium rate (3/20 (15％)),the temporary hypoparathyroidism rate (2/20 (10％)) and the decline degree of parathyroid hormone (47.3 ± 14.31)％ in the carbon nanoparticles group were lower than those of the control group(P ＜0.05).The number of central lymph node dissected (9.45 ± 2.33) pieces/case in the carbon nanoparticles group was more than that of the control group (P ＜ 0.05).The number of lymph node in the right recurrent laryngeal nerve (3.12 ± 0.65) pieces/case was more than that of the control group(P ＜ 0.05).The lymph node metastasis rate had no significant difference between the 2 groups(P ＞ 0.05).The incidence of postoperative complications in the carbon nanoparticles group was lower than that of the control group (P ＜ 0.05).Conclusion The application of carbon nanoparticles in total thyroidectomy combined with central lymph node dissection can contribute to the recognition and protection of parathyroid glands and its blood supply,improve the rate of central lymph node dissection,and reduce the incidence of postoperative complications.

Animals ; Carcinogens ; toxicity ; Female ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Sensitivity and Specificity ; Urethane ; toxicity

Objective To analyze the effect of N-acetyl-L-cysteine(NAC)on endoplasmic reticulum stress mediated HepG2 cells apoptosis and evaluate the role of NAC in the treatment of liver injury.Methods HepG2 cells were treated with thapsigargin(TG)to establish the model of oxidative endoplasmic reticulum stress mediated apoptosis,and NAC was used to intervene in apoptosis.To evaluate the apoptosis,various methods such as MTT assay,flow cytometry,DNA ladder and Western blot were performed.Results After treated with 2 μmol/L TG for 0,24,36 and 48 hours,the vitality of HepG2 cells decreased.The ratio of apoptotic cells increased along with the prolonged treatment duration of TG(0.7%±0.5%,27.6%±6.3%,29.7%±3.3%,47.9%±3.5% respectively,P<0.05),and the production of reactive oxygen species(ROS)also increased in time-dependent manner(14.0%±0.5%,36.1%±3.0%,38.2%±6.0%,48.3%±12.4%,P<0.05).The HepG2 cells showed typical morphologic change of endoplasmic retieulum stress induced by 2 μmol/L TG after 36 h and 48 h.DNA ladder was observed at the same concentration and time point correspondingly.Endoplasmic reticulum stress mediated-apoptosis was confirmed by Western blot.Both 10 mmol/L and 20 mmol/L NAC could protect ceils from apoptosis.The ratio of apoptotic cells decreased to 14.0%±1.3% and 11.0%±0.3%,respectively.The production of ROS decreased to 34.7%±0.8% and 31.5%±2.9%,respectively.The effect was related to the concentration of NAC.Conclusions As a Ca2+-adenosine triphoshatase inhibitor,TG may disrupt intracellular calcium homeostasis,which can induce endoplasmie reticulum stress and apoptosis.NAC,the precursor of the synthesis of-SH,can directly inhibit the ROS reaction and alleviate liver damage,which may play a role in the treatment of liver failure.

Objective To study the process and mechanism underlying Wallerian degeneration of the central nervous system after stroke.Methods A case suspected of stroke with bilateral symmetrical lesions in middle cerebellar peduncles (MCPs) was described.The etiology of bilateral MCP abnormal signals on MR was analyzed according to the clinical process and neuroanatomy.Results Unilateral paramedian pontine infarction,covering the crossing area of pontocerebellar fibers, would cause bilateral secondary degeneration of MCPs,with hyperintense signals on T2-,Flair and diffusion-weighted images.Conclusions Wallerian degeneration of projecting system is a common sequel after stroke and should not be misdiagnosed as other diseases.

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

Objective To evaluate the left ventricular synchrony after myocardial infarction (MI) by ultrasound targeted microbubbles destruction (UTMD)-mediated angiogenin 1 (Ang1) gene transfection in canine.Methods Twenty-one dogs were divided into three groups (n =7 in each group):①control group (healthy dogs);②MI group (MI dogs without treatment);③UTMD group (MI dogs with UTMD treatment).One month later,the size and systolic function of heart were measured by echocardiography.The synchronization parameters derived from two dimensional-speckle tracking imaging(2D-STI) included the standard deviation and maximum difference of time to peak strain for all left ventricular segments (Tls-SD,Trs-SD,Tcs-SD,Tls-Dif,Trs-Dif and Tcs-Dif).CD31 and α-SMA were applied for quantifying capillary and arteriolar density.The Ang1,SERCA2a and PLB protein were detected by Western blotting.Results ① One month later,the conventional ultrasonic parameters were compared among three groups,the LVEDD,LVESD and E/e'increased and LVEF,e'and E/A reduced in MI group than those in control group,all of them partially recovered in UTMD group than those in MI group,but were still lower than those in control group (P ＜0.05);②The left ventricular synchrony parameters of Tls-SD,Tls-Dif and Trs-SD showed significant differences among the three groups(P ＜0.05),the degree of dyssynchrony increased in MI group than control group,they were lower in UTMD group than those in MI group.The Tcs-SD,Tcs-Dif and Trs-Dif showed no significant difference among three groups (P ＞ 0.05);③ The immunohistochemistry showed the higher blood vessel density in UTMD group than that in MI group(P ＜ 0.05);④The relative quantity of Ang1 was significantly higher in UTMD group.The relative quantity of SERCA2a protein was lower in MI group than that in control group,increased in UTMD group,the trend of PLB was contrary to it.The differences were statistically significant (all P ＜0.05).Conclusions The UTMD-mediated Ang1 gene transfection can promote angiogenesis after MI,reverse left ventricular remodeling and improve left ventricular synchrony.The myocardial synchrony may be related to the expression of calcium ions key protein SERCA2a and PLB.

BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

OBJECTIVETo investigate the role of transcript factor SCL/TAL-1 gene in the erythroid differentiation through the knockdown of SCL/TAL-1 mRNA by RNA interference.

METHODSThe plasmid of pTRIP-dU3-RNAiTALh-EF1a-GFP with SCL/TAL1 shRNA was transfected into EPO-induced K562 cell line with erythroid differentiation via lentiviral vector system and the expression of SCL/TAL-1 mRNA decreased. The plasmid pTRIP-dU3- RNAiluc-EF1-GFP expressing EGFP gene was as control. The mRNA levels of SCL/TAL-1 and erythroid related RhD, GPA, CD47 in the cell lines were detected by RT-PCR, and erythroid antigen CD71, CD235a were examined by flow cytometry.

RESULTS(1) After 48 h of transfect, more than 95% of K562 cells were GFP positive, indicating the infection rate of the plasmids in the K562 cells more than 95%. (2) The results of RT-PCR showed SCL/TAL-1 mRNA expression in the K562 cell line of knockdown of SCL/TAL-1 was significantly lower than that in the control (P < 0.05). The mRNA levels of CD47 and RhD were also significantly lower, however, GPA decreased slightly in comparison with the control. (3) The expressions of CD71 and CD235a markedly reduced in the K562 cell line of knockdown of SCL/TAL-1 with positive rates as 10.4% and 76.5%, while the positive rates in the control as 94.3% and 83.6%.

CONCLUSIONOur findings suggested that transcription factor SCL/TAL-1 might play an positive role in erythroid differentiation.

Cell Differentiation ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; K562 Cells ; RNA Interference

OBJECTIVETo investigate the anti-leukemia effect of oridonin on Ph(+) acute lymphoblastic leukemia (ALL) cell line SUP-B15.

METHODSHuman Ph(+) ALL cell line was cultured in vitro. The 50% inhibition concentration (IC(50)) of oridonin against SUP-B15 cell line was examined using modified MTT assay. The cellular morphologic changes were observed using a light microscope. The percent of apoptosis of SUP-B15 cell line after drug treatment was evaluated by flow cytometric analysis. The active levels of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK, STAT5 signaling pathways and the expression levels of Bcl-2 and BAX were examined by Western blot.

RESULTSOridonin inhibited the growth of SUP-B15 cell line in both time- and dose-dependent manner with the IC(50) of oridonin as (7.08 ± 1.21) µmol/L after 72 h treatment. The cellular membrane of SUP-B15 cell line treated with oridonin became unsharp, some of them disintegrated. Oridonin induced apoptosis in SUP-B15 cell line with the apoptosis rates following 0, 5, 10 µmol/L oridonin treatment for 24 h were (6.67 ± 0.83)%, (18.30 ± 1.79)% and (37.63 ± 7.12)%, respectively. Oridonin inhibited activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, which were constitutively activated in SUP-B15 cell line, down-regulated the level of anti- apoptotic protein Bcl-2 and up-regulated the expression of pro-apoptotic protein Bax.

CONCLUSIONOridonin exerted anti-leukemia effect in Ph(+)ALL cell line SUP-B15 by inhibiting the activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, down-regulating the expression of Bcl-2 and up-regulating the expression of BAX.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Kaurane ; pharmacology ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Signal Transduction

Objective To assess the clinical application of contrast-enhanced MR angiography using three-dimensional(3D)time-resolved imaging of contrast kinetics(CE-MRA 3D-TRICKS).Methods TRICKS is a high temporal resolution(2—6s)MR angiographic technique using a short TR(2.8— 4.0 ms)and TE(0.9—1.3 ms),partial echo sampling and the central part of the k-space being updated more frequently than the peripheral part of the k-space.Pre-contrast mask 3D images are first acquired and 15--20 sequential 3D images following bolus injection of Gd-DTPA are then acquired.Results Thirty patients underwent contrast-enhanced MR angiography using TRICKS.Twelve vertebral arteries were well displayed on TRICKS.Seven of them showed normal,bilateral vertebral artery stenosis was shown in 1 case, and unilateral vertebral artery stenosis was shown in 4 wth aecompaning ipsilateral carotid artery bifurcation stenosis in one case.Bilateral renal artery showed normal in 4 cases,and the artery in transplanted kidney showed normal in one case and stenosis in another case.The cerebral artery showed normal in 2 cases, sagittal sinus thrombosis was detected in one case and intracranial arteriovenous malformation in one case. Pulmonary artery displayed normal in 3 cases,pulmonary artery thrombosis was seen in one case and pulmonary sequestration's abnormal feeding artery and draining vein was revealed in one case.The feeding artery in left lower limb fibrolipoma was showed in one case.The radial-ulnar artery artificial fistula stenosis was seen in one case,and left antebrachium hemangioma was showed in one case.Conclusion TRICKS can clearly delineate the whole body vascular system and can reveal any vascular abnormality.It is convenient and with high successful rate,which make it the first method of choice in displaying vascular abnormality.