Head and Neck Neoplasms

The AC impedance of human hepatoma SMMC-7721 cells were measured in our laboratory by Agilent 4294A impedance analyzer in the frequency range of 0.01-100 MHz. And then the effect of hematocrit on electrical impedance characteristics of hepatoma cells was observed by electrical impedance spectroscopy, Bode diagram, Nyquist diagram and Nichols diagram. The results showed that firstly, there is a frequency dependence, i.e., the increment of real part and the imaginary part of complex electrical impedance (δZ', δZ"), the increment of the amplitude modulus of complex electrical impedance (δ[Z *]) and phase angle (δθ) were all changed with the increasing frequency. Secondly, it showed cell volume fraction (CVF) dependence, i. e. , the increment of low-frequency limit (δZ'0, δ[Z*] 0), peak (δZ"(p), δθ(p)), area and radius (Nyquist diagram, Nichols diagram) were all increased along with the electric field frequency. Thirdly, there was the presence of two characteristic frequencies: the first characteristic frequency (f(c1)) and the second characteristic frequency (f(c2)), which were originated respectively in the polarization effects of two interfaces that the cell membrane and extracellular fluid, cell membrane and cytoplasm. A conclusion can be drawn that the electrical impedance spectroscopy is able to be used to observe the electrical characteristics of human hepatoma cells, and therefore this method can be used to investigate the electrophysiological mechanisms of liver cancer cells, and provide research tools and observation parameters, and it also has important theoretical value and potential applications for screening anticancer drugs.
Carcinoma, Hepatocellular ; Cell Line, Tumor ; Cell Membrane ; Cytoplasm ; Electric Impedance ; Electrophysiological Phenomena ; Humans ; Liver Neoplasms ; Membrane Potentials

BACKGROUND: SSh as a Hedgehog signal protein can promote bone development, growth and remodeling.OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) combined with Shh modified nano-hydroxyapatite/collagen (nHAC) in the repair of femoral defects in rats.METHODS: Forty-eight Sprague-Dawley rats were randomly divided into four groups, and the model of femoral defects was established in these rats. At 14 days after modeling, experimental group was implanted with the BMSCs/Shh modified nHAC, scaffold group was implanted with simple nHAC, cell scaffold group was implanted with BMSCs/nHAC,and blank control group was without any implantation. At 3, 6, 9, 12 weeks after repair, X-ray examination, bone density measurement and bone biopsy in bone defect area were performed.RESULTS AND CONCLUSION: (1) X-ray examination: The Lane-Sandhu X-ray score and bone mineral density value in the experimental group at different time points after operation were significantly higher than those in the other three groups (P < 0.05). (2) Hematoxylin-eosin staining: 12 weeks after repair, a small amount of bone tissues but no bone marrow formed in the scaffold group; a small amount of bone tissues with absence of bone marrow formed in the cell scaffold group, and the residual scaffold was visible; in the experimental group, the scaffold was completely absorbed,and mature bone and medullary cavity formed with presence of bone marrow. (3) Scanning electron microscope observation: 12 weeks after repair, irregular arrangement of bone fibers and a large number of bone fossae were observed in the scaffold group; the cell scaffold group showed a large number of osteoblasts, but bone fibers still arranged irregularly; in the presence of the Haversian system, a large number of regularly arranged bone trabeculae were detective in the experimental group. These results elucidate that the Shh modified nHAC/BMSCs complex can promote the repair of bone defects.

Objective Through the analysis of cases,to improve the cognition of clinicians on extracranial vas-cular embolism diseases(VED)in children.Methods The clinical information included incidence trend,thrombophi-lia,clinical features,treatment and prognosis of 29 children with VED,who were admitted from January 201 1 to Decem-ber 201 5 in Nanjing Children′s Hospital Affiliated to Nanjing Medical University,and retrospectively analyzed.Results The annual rate of VED increased from 0 to 1 .49 per 1 0 000 hospital admissions from January 201 1 to December 201 5.In 29 cases,there were 20 male and 9 female,with an average age of 6.03 years old (2 h -1 3.83 years old).In-fants accounted for >20%(6 /29 cases).The average age of the female,non infected group,rheumatic disease group and venous thrombosis group was higher than that of the male,the infection group,the non rheumatic disease group and the arterial embolization group,but there was no significant difference(all P >0.05).Thrombophilia were as follows:in-fection disease,heart disease,kidney disease syndrome,rheumatoid disease,tumor,and twins embolization syndrome. Risk factors included central venous line,immobility,dehydration,glucocorticoids use,etc.Heart disease was the leading cause of non thrombotic embolism,while other thrombophilic diseases resulted in thrombotic embolism.Pulmonary em-bolism was mainly seen in patients with severe pneumonia,especially mycoplasma pneumoniae pneumonia (MPP ).The older the age,the more typical clinical manifestations,and significantly elevated D dimer,fibrin/fibrinogen degradation products tips VED.Treatment depends on the degree of risk.Thirty -seven point five percent(3 /8 cases)of children with arterial embolism were less than 1 year old,and the mortality 25.00% (2 /8 cases)was significantly higher than that of venous thrombosis 1 7.65%(3 /1 7 cases)and 1 1 .76% (2 /1 7 cases).Conclusion Heart disease mainly cause arterial embolism and infection,while nephritic syndrome and rheumatic disease mainly cause venous embolism.Pulmona-ry embolism is mainly seen in children with severe pneumonia,especially MPP.The risk of arterial embolization is higher than that of venous embolism.

BACKGROUND: Polylactic acid copolymer bone scaffold has excellent biodegradability, and it is easy to be shaped and can promote the formation and growth of bone tissue and blood vessel.OBJECTIVE: To observe the effects of adipose-derived stem cells(ADSCs)/poly(lactic-co-glycolic acid) (PLGA) complex on the biomechanical properties after fracture healing in osteoporotic bone.METHODS: Sixty Sprague-Dawley rats were randomly divided into four groups: blank control group received no treatment; the bilateral tibial fracture model was made after 3 months of bilateral ovarian resection in model group; the bilateral tibial fracture model was made and ADSCs were implanted into the bone after 3 months of bilateral ovarian resection in cell therapy group; the bilateral tibial fracture model was made and the PLGA/ADSCs complex was implanted after 3 months of bilateral ovarian resection in combined treatment group.The bone mineral density, callus thickness, biomechanical parameters and the microstructure of the trabecular bone were detected.RESULTS AND CONCLUSION: (1) The bone density: The bone density of the model group was significantly lower than that of the blank control group (P < 0.05); the bone mineral density of the cell therapy group and the combined treatment group was higher than that of the model group (P < 0.05), but lower than that of the control group (P < 0.05); and the bone mineral density of the combination treatment group was higher than that of the cell therapy group (P < 0.05). (2)Thickness of the callus: The thickness of the callus in the cell therapy group and combined treatment group was higher than that of the model group and blank control group (P < 0.05); moreover, the thickness of the callus in the combined treatment group was higher than that of the cell therapy group (P < 0.05). (3) Biomechanical test: The failure load, stress and shear strength, elastic modulus were decreased in the model group compared with the blank control group (P < 0.05), while the shear strain increased (P < 0.05). Compared with the model group, the failure load, ultimate stress, shear strength, elastic modulus were increased in the cell therapy group and combined treatment group (P < 0.05), and the shear strain was decreased (P < 0.05). Moreover, the combined treatment group showed more changes in these biomechanical parameters (P < 0.05). (4) The trabecular bone microstructure: The model group presented with trabecular derangement, spacing increases, and even fracture and lacuna. After ADSCs or ADSCs/PLGA transplantation,the trabecular bones increased in number, thickness, and spacing, and the number of lacunae reduced. In conclusion,ADSCs combined with PLGA in the treatment of osteoporotic fracture can significantly improve the biomechanical parameters of bone tissue after healing.

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism

Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression Profiling ; Genomics ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Metastasis ; genetics ; Neoplasm Recurrence, Local ; genetics

Objective To study the effect of protein C mutation on venous thrombosis (VTE) by PROC gene sequencing in patients with VTE. Methods Human PROC gene sequence was designed to amplify the third exon region of the PC-Gla domain , and then the PCR products were sequenced to search for a single nucleotide mutation (SNVs). The SNVs was constructed into eukaryotic expression system and a stable expression of wild-type and mutant PC cells were also constructed. At the same time , the distribution of PC levels in normal and VTE patients were detected with ELISA. Results Three single nucleotide mutations were found in different patients. In HUVEC cells, the synthesis of PC decreased in each mutant strain. The PC level in the normal patients , VTE patients , and the mutant samples were detected , which were significantly lower in the mutant samples than that of the VTE group (P = 0.035 3) and the normal level (P < 0.000 1). Conclusion Three mutation sites PCArg-1Cys , PCVal34Met and PCArg9Cys are important genetic factors lead to a significant decrease in plasma PC levels and the increase of VTE risk.

Objective To establish binomial equation Cole-Cole mathematical modeling methods and the parameters in rat erythrocyte impedance spectra,as well as to discuss the sources of the ? and the ? dispersion.Methods The impedance of erythrocyte suspension was measured with Agilent 4294A impedance analyzer within the frequency range of 0.01~100 MHz.The residual error of the fitting curve was determined with non-linear numerical calculation of Cole-Cole equation.Then the Cole-Cole mathematical model parameters of rat erythrocyte were established with the Bode and the Nyquist plots.Results The Cole-Cole mathematical model parameters of rat erythrocyte impedance spectrum: the high-frequency limit of impedance R∞=63/?cm,the 1st relaxation impedance increment ?R1=59/?cm,the 1st characteristic frequency fC1 =2.4 MHz,the 1st relaxation dispersion angle ?1=0.07,the 2nd relaxation impedance increment ?R2=48/?cm,the 2nd characteristic frequency fC2 =370 MHz,the 2nd relaxation dispersion angle ?2=0.22.Conclusion The ? and ? relaxation behavior of rat erythrocyte impedance spectrum can be expressed with the two-type Cole-Cole mathematical model.The ? dispersion is formed from the capacitive response of cell membrane and the ? dispersion from frequency response of intracellular hemoglobin.

Objective To investigate whether a novel long-acting tumor necrotic factor (TNF) antagonist (soluble TNF receptor:IgG Fc [sTNFR:IgG-Fc]) can protect hepatocyte damage against liver failure caused by drugs in immunity-induced cirrhotic rats.Methods Wistar rats were repeatedly sensitized by human serum albumin (HSA) emulsified in complete freud adjuvant.The blood was collected at day 10 after the final sensitization.If anti-albumin antibody was positive,the rats were intravenously injected with HSA twice a week.After six weeks,liver cirrhosis was induced by immunity.All the model rats were divided into three groups with 15 each.Liver failure was induced with D-galactosamine/ lipopolysaccharide (LPS) intraperitoneal injection in the rats with liver cirrhosis in model group.The rats in pretreatment group were intraperitoneally injected with long-acting soluble TNF receptor p55 18 h before D-galactosamine/LPS injection.The control group were injected with 0.9％ sodium chloride.General condition,survival rate,liver function and pathological changes were all examined.Serum levels of interleukin (IL)-6,IL-22 and intrahepatic level of IL-6 were detected by enzyme linked immunosorbent assay (ELISA).The activity of Caspase 3 in hepatocyte lysis solution was measured by spectrophotography.Real-time polymerase chain reaction (PCR) was used to detect mRNA expressions of proliferating cell nuclear antigen (PCNA),bcl-2,bax and IL-22 receptor.Data were analyzed by variance analysis among groups.Results Rats in model group were dispirited with poor response after 12 hours and only 3 survived,compared with soluble TNF receptor p55 pre-treated group rats,in which all survived (P=0.029 8) with flexible response.Serum alanine aminotransferase levels in these two groups were (6 533± 360) and (105 ± 7) U/L,respectively.Hepatic regenerative nodule developed massive or submassive necrosis with septal fibrosis in model group,whereas soluble TNF receptor p55 alleviated the inflammatory and necrosis reaction of hepatic tissue.Serum IL-6 levels in model group and pretreatment group were (842.0±12.9) and (91.9±1.6) pg/mL,respectively (F=380.30,P＜0.01).Intrahepatic levels of IL-6 in these two groups were (26.2±1.2) and (11.1±0.8) pg/mL,respectively (F=176.90,P＜0.01),and serum IL-22 levels were (167.0±27.8) and (988.0±109.6) pg/mL,respectively (F=37.91,P＜0.01).Hepatic Caspase-3 activity was reduced by almost 60％ by soluble TNF receptor p55 pretreatment (F=303.70,P＜0.01) and bax expression reduced by 22％ (F=108.80,P＜0.01),while bcl-2 and PCNA expressions were up-regulated by 3.6-folds and 23.0-folds,respectively (F=115.60,P＜0.01; F=594.20,P＜0.01).Conclusions Long acting soluble TNF receptor p55 could improve survival rate,liver function and reduce inflammatory reaction of rats with liver failure induced by drugs on the basis of liver cirrhosis caused by immunity,which indicates that this drug may process a potential therapeutic value.