Human adipose-derived mesenchymal stem cells and synovial-derived mesenchymal stem cells synergistically inhibit the degeneration of inflammatory chondrocytes / 中国组织工程研究

BACKGROUND: Intra-articular injection of cells targeting to change the microenvironment in lesions can act on early osteoarthritis of inflammatory chondrocytes. Implanted cells affect the progress of the disease by the cell characteristics. OBJECTIVE: To explore the synergistic effect of mesenchymal stem cells from human knee adipose (ADMSCs) and synovial tissues (SDMSCs) to inhibit the degeneration of inflammatory chondrocytes. METHODS: ADMSCs, SDMSCs and inflammatory chondrocytes were primary cultured. Under in vitro two-dimensional culture conditions, cell proliferation assay (MTS) was performed to detect the proliferation of three kinds of cells. Differences in chondrogenic markers at mRNA and protein levels between three kinds of adherent cells were detected by quantitative PCR and immunofluorescence. Under in vitro three-dimensional mixed culture conditions, three groups were set up: (1) ADMSCs+inflammatory chondrocytes (A+C group), (2) SDMSCs+inflammatory chondrocytes (S+C group), and (3) ADMSCs-SDMSCs+inflammatory chondrocytes (A+S+C group). Alcian blue staining, safranin O staining and type Ⅱ collagen immunohistochemistry staining were performed on the mixed-cultured cell mass paraffin sections followed by quantitative analysis. Chondrogenic differentiation in each group was detected by quantitative PCR. Culture supernatants were collected to detect the secretion of pro-inflammatory and anti-inflammatory factors by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: Under the two-dimensional culture, the proliferative rate of ADMSCs was significantly higher than that of inflammatory chondrocytes and SDMSCs (P < 0.05). The expression of type Ⅱ collagen mRNA and protein and proteoglycan protein in inflammatory chondrocytes was significantly higher than that in the other two kinds of cells (P < 0.01). Under the three-dimensional culture, the percentage of chondrogenic area per total area was significantly higher in the A+S+C group than the S+C and A+C groups (P < 0.05). The expression of type Ⅱ collagen and proteoglycan was significantly higher in the A+S+C group than the S+C and A+C groups (P < 0.05). Compared with the other two groups, the S+C group showed higher levels of interleukin 1, interleukin 6, and tumor necrosis factor α, but lower level of interleukin 10 (P < 0.05). To conclude, the combined use of ADMSCs and SDMSCs synergistically inhibits the degeneration of inflammatory chondrocytes.

Effect of Ding's herb enema prescription on intestinal tissue relateddifferences target in rat colitis using protein array technology / 中国药理学通报

Aim To investigate the effect of Ding''s herb enema prescription on intestinal tissue related target in rat colitis induced by dextran sulfate sodium(DSS), and to elucidate the mechanism of Ding''s herb enema prescription in improving the intestinal inflammation and intestinal fibrosis.Methods Rats were fed with 3.5％ DSS.The rats were randomly divided into positive drug group, model group, Control group, and Ding''s herb enema prescription group.The positive drug group was treated with mesalazine enema, and Ding''s herb enema prescription group was treated with Ding''s herb enema prescription.The colon mucosa was taken once a day for 6 weeks.The changes of intestinal inflammatory response and intestinal fibrosis related proteins were detected by GSR-CAA-67 antibody protein array, and the differentially expressed proteins were screened out.Results Eight proteins showed statistical differences, including IFN-γ, erythropoietin(EPO), TIMP-2, TIM-1, IL-6, TIMP-1, TNF-α, IL-22 (P0.05).Conclusions Ding''s herb enema prescription has the effect of multiple targets, which may improve the intestinal inflammatory response and intestinal fibrosis to achieve the purpose of treatment of ulcerative colitis(UC).

Cytotoxic effects of local anesthesia through lidocaine/ropivacaine on human melanoma cell lines / Efeitos citotóxicos de anestesia local com lidocaína/ropivacaína em linhagens celulares de melanoma humano

Abstract Background: Local anesthetics (LAs) are generally considered as safe, but cytotoxicity has been reported for several local anesthetics used in humans, which is not well investigated. In the present study, the cytotoxicity of lidocaine, ropivacaine and the combination of lidocaine and ropivacaine were evaluated on human melanoma cell lines. Melphalan, a nitrogen mustard alkylating agent, was used as a control agent for comparison of cytotoxic activity. Methods: Melanoma cell lines, A375 and Hs294T, were exposed to 1 h to different concentrations of above agents. Cell-viability after exposure was determined by flow cytometry. Results: Investigated LAs showed detrimental cytotoxicity on studied melanoma cell lines in time- (p < 0.001), concentration- (p < 0.001), and agent dependant. In both A375 and Hs294T cell lines, minimum cell viability rates were found after 72 h of exposure to these agents. Lidocaine 2% caused a reduction of vital cells to 10% ± 2% and 14% ± 2% in A375 and Hs294T, respectively after 72 h of exposure. Ropivacaine 0.75% after 72 h reduced viable cells to 15% ± 3% and 25% ± 3% in A375 and Hs294T, respectively. Minimum cell viability after 72 h exposure to the combination was 10% ± 2% and 18% ± 2% in A375 and Hs294T, respectively. Minimum cell viability after 72 h exposure to melphalan was 8% ± 1% and 12% ± 2%, in A375 and Hs294T, respectively. Conclusion: LAs have cytotoxic activity on human melanoma cell lines in a time-, concentration- and agent-dependant manner. Apoptosis in the cell lines was mediated through activity of caspases-3 and caspases-8.

Resumo Justificativa: Os anestésicos locais (ALs) são geralmente considerados como seguros, mas citotoxicidade foi relatada em vários anestésicos locais usados em seres humanos, a qual não é bem investigada. No presente estudo, a citotoxicidade de lidocaína e ropivacaína e da combinação de lidocaína e ropivacaína foi avaliada em linhagens celulares de melanoma humano. Melfalano, um agente alquilante de mostarda nitrogenada, foi usado como um agente de controle para a comparação da atividade citotóxica. Métodos: Linhagens celulares de melanoma, A375 e Hs294T foram expostas por uma hora a concentrações diferentes dos agentes mencionados acima. A viabilidade celular após a exposição foi determinada por citometria de fluxo. Resultados: Os ALs investigados mostraram citotoxicidade prejudicial nas linhagens celulares de melanoma estudadas dependente do tempo (p < 0,001), da concentração (p < 0,001) e do agente. Em ambas as linhagens de células A375 e Hs294T, níveis mínimos de viabilidade celular foram encontrados após 72 horas de exposição a esses agentes. Lidocaína a 2% provocou uma redução das células vitais para 10% ± 2% e 14% ± 2% em A375 e Hs294T, respectivamente, após 72 horas de exposição. Ropivacaína a 0,75% após 72 horas reduziu as células viáveis para 15% ± 3% e 25% ± 3%, em A375 e Hs294T, respectivamente. A viabilidade celular mínima após exposição de 72 horas para a combinação foi de 10% ± 2% e 18% ± 2% em A375 e Hs294T, respectivamente. A viabilidade celular mínima após exposição de 72 horas ao melfalano foi de 8% ± 1% e 12 ± 2, em A375 e Hs294T, respectivamente. Conclusão: Os ALs têm atividade citotóxica em linhagens de celulares de melanoma humano de modo dependente do tempo, da concentração e do agente. A apoptose nas linhagens celulares foi mediada por meio da atividade das caspases-3 e caspases-8.

The influence of microcystin-LR on monocytes and lymphocytes of mice / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the influence of microcystin-LR (MC-LR) on monocytes and lymphocytes in blood of mice and to find a sensitive index of toxic effects.</p><p><b>METHODS</b>Specific pathogen free Kunming male mice, aging 1 month-old,were randomly divided into 5 groups by weights, 7 mice for each group. The mice in 5 groups were exposed to MC-LR through intraperitoneal injection at 0, 3.125,6.250, 12.500 and 25.000 µg/kg respectively for 7 days. Then cytokine levels in the serum were measured by radioimmunoassay, DNA-protein crosslinks (DPC) was measured by the SDS/KCl precipitation technique, and the phagocytosis and ROS of leukocytes were detected by flow cytometry.</p><p><b>RESULTS</b>The levels of interleukin 6 in the 6.250, 12.500 and 25.000 µg·kg(-1)·d(-1) dose groups were (346.837 ± 25.536), (360.847 ± 37.886) and (434.245 ± 35.858)pg/ml respectively, which were significantly higher than those in the control group which the value was (232.775 ± 32.816) pg/ml (t values were -7.258, -6.760 and -10.966 respectively, P values were all < 0.05).While the level of tumor necrosis factor-alpha was(10.782 ± 0.966) fmol/ml in 25 µg·kg(-1)·d(-1) dose group was statistically lower than it in the control group which the value was (16.878 ± 3.378) fmol/ml (t value was 4.591, P < 0.05). The DPC levels of lymphocytes in 6.250, 12.500 µg·kg(-1)·d(-1) dose group were (242.576 ± 7.545),(241.472 ± 2.793) ng/ml,higher than it in the control group while the value was (228.657 ± 4.130) ng/ml (t value was -4.282, -6.801, P values were all <0.05). The fluorescence intensity of DCF in lymphocytes in the 4 treated groups were separately 3299.37 ± 120.54, 3281.38 ± 58.34, 3308.06 ± 136.12 and 3346.92 ± 108.69, all significantly lower than 3770.81 ± 131.39 in the control group (t values were 6.995, 9.007, 6.472 and 6.577 respectively, and P values were all <0.05). The fluorescence intensity of DCF in monocytes in the 4 treated groups (3271.51 ± 140.79, 3270.05 ± 117.92, 3326.90 ± 114.39 and 3292.49 ± 145.97 respectively) were also significantly lower than the value in the control group was 3841.72 ± 130.92 (t values were 7.847, 8.584, 7.835 and 7.411 respectively, P values were all <0.05). There was no significant difference in other index among the four experiment groups and the control group.</p><p><b>CONCLUSION</b>The MC-LR administered via intraperitoneal injection to mice induced the alterations of some cytokines of monocytes and lymphocytes in blood. By comparison, the ROS of leukocyte was the most sensitive index.</p>

Clinical utilization of microembolus detection by transcranial Doppler sonography in intracranial stenosis-occlusive disease / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To discuss the clinical ultiliazation and significance of microembolus detection by transcranial Doppler (TCD) sonography in intracranial stenosis-occlusive disease.</p><p><b>DATA SOURCES</b>All related articles in this review were mainly searched from PubMed published in English from 1996 to 2012 using the terms of microembolic signal, transcranial Doppler, intracranial stenosis, stroke.</p><p><b>STUDY SELECTION</b>Original articles and reviews were selected if they were related to the clinical utilization of microembolus detection in intracranial stenosis-occlusive disease.</p><p><b>RESULTS</b>Intracranial stenosis is a significant cause of cerebral emboli, and microembolus detection by TCD sonography were widely used in exploring the mechanisms of ischemic stroke with intracranial stenosis (including the middle cerebral artery stenosis and the vertebral-basilar stenosis), evaluating the prognosis of acute stroke, evaluating the therapeutic effects, and predicting the recurrent events of stroke.</p><p><b>CONCLUSION</b>Microembolus detection by TCD sonography plays an important role in the cerebral ischemic stroke patients with intracranial stenosis.</p>

The establishment and application of the method with virus concentration and detection in drinking water / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study aimed to construct an effective method to concentrate and detect virus in drinking water, and human adenovirus pollution status in actual water samples was monitored by constructed method.</p><p><b>METHODS</b>The concentration efficient of NanoCeram filter for the first concentration with source water and drinking water and the concentration efficient of the different concentrations of PEG 8000 for the second concentration were assessed by spiking f₂ bacteriophage into water samples. The standard of human adenovirus for real-time PCR was constructed by T-A clone. The plasmid obtained was identified through sequence analyzing and consistency check comparing to target gene fragment was conducted by using blast algorithm. Then, real-time PCR was constructed to quantify the concentration of human adenovirus using the plasmid as standard. Water samples were concentrated by using NanoCeram filter on the spot and then concentrated for the second time by PEG/NaCl in 2011. The DNA of concentrated samples were extracted for the quantification of human adenovirus in real-time PCR subsequently to monitor the pollution of human adenovirus in water.</p><p><b>RESULTS</b>For the first concentration by NanoCeram filter, the recovery rates were (51.63 ± 26.60)% in source water and (50.27 ± 14.35)% in treated water, respectively. For the second concentration, the highest recovery rate was reached to (90.09 ± 10.50)% at the concentration of 0.13 kg/L of PEG 8000. The sequence identity score of standard of adenovirus for real time PCR and adenovirus gene was 99%, implying that it can be successfully used to quantification with human adenovirus. The levels of human adenovirus in the water samples sampled in 2011 ranged from 4.13×10³ to 2.20×10⁶ copies/L in source water, while range from 5.57×10² to 7.52×10⁵ copies/L in treated water and the removal efficiency range was (75.49 ± 11.71)%.</p><p><b>CONCLUSION</b>NanoCeram filers combined with PEG/NaCl was an effective method to concentrate virus in aquatic environment. There was a large number of human adenovirus in source water, and it is not sufficient to remove them thoroughly through conventional water treatment processes.</p>

Assessment of left ventricular function and synchrony in patients with coronary disease after PTCA and stent implantation by quantitative tissue velocity imaging / 中华超声影像学杂志

Objective To evaluate the left ventricular function and the synchrony of myocardial ischemic segments in patients with coronary heart disease (CHD) after PTCA and stent implantation by quantitative tissue velocity imaging (QTVI). Methods Thirty-six patients with isolated left anterior descending stenosis (≥75%) were examined by QTVI three days before, one week and one month after successful PTCA and stent implantation to measure the following items of 5 different left ventricular segments: peak systolic velocity( Vs), early diastolic velocity (Ve), late diastolic velocity (Va), time to peak systolic velocity(Ts). Then the coefficient of variation (SD/mean) of the 5 different Ts were calculated.Results The value of Vs,Ve and Va were decreased and the Ve/Va ratio was reverses three days before PTCA + stent. Compared with that before PTCA + stent,the value of Vs and Ve were increased significantly in one week ( P ＜0. 05) and one month( P ＜0.01 ) after PTCA + stent,respectively,the value of Va was not statistically significant. Ve/Va ratio was recovered in one week after PTCA treatment. Ts and Ts-SD were shorted dramatically in one week( P ＜0. 05) and one month( P ＜0.01 ) after PTCA + stent compared with that before PTCA + stent in which Ts were prolonged more than 33 ms. Conclusions QTVI can quantitatively assess the left ventricular function and the synchrony of myocardial ischemic segments, and can be used to real-time detect the changes of function and synchrony of left ventricle after PTCA and stent implantation.

Diagnostic value on finger parenchyma tumor by ultrasonography / 中华超声影像学杂志

Objective To explore the clinical diagnostic value of high frequency ultrasound on finger parenchyma tumor. Methods Twenty-two patients with finger parenchyma tumor were analyzed retrospectively by clinical and ultrasonographic characteristics including location, size, shape, echo, and color flow signals of tumors. Those findings were compared with pathological data after surgery. Results Out of 22 parenchyma tumor patients, tendosynovial giant cell tumor were present in 7 patients, fibrous tumor in 3 patients,glomus tumor in 5 patients, and ganglia in 7 patients. Differences between ultrasonographic appearances and pathologic features were found in parenchyma tumor. The tendosynovial giant cell tumors were demonstrated plentiful color flow signals and heterogeneous echoic mass without capsules. There was not found color flow signal but an intact membrane in tendosynovial fibrous tumors. Glomus tumors were hypoechoic with an intact membrane, abundant color Doppler signals, and Ⅲ level was classified by Alder. Ganglia were expressed in cystic structure. Conclusions Characteristic features of ultrasonographic appearance were found in different kind of parenchyma tumors. High frequency ultrasound is an effective method to diagnosis finger parenchyma tumor.

Correlations of ambulatory pulse pressure to large artery elasticity and vascular endothelial function in patients with primary hypertension / 中国组织工程研究

BACKGROUND: Ambulatory blood pressure monitoring can sensitively and objectively reflect blood pressure level, which is closely related to target organ damage and disease prognosis. In hypertension, vascular endothelial damage is the most common lesion to target organs. There is little known about how ambulatory pulse pressure correlates to large artery elasticity and vascular endothelial function. OBJECTIVE: To investigate changes of large artery elasticity and of vascular endothelial function in patients with primary hypertension using an automatic pulse wave velocity determinator and ultrasound techniques, and to analyze the correlation of ambulatory pulse pressure to large artery elasticity and vascular endothelial function.DESIGN: A non-randomized concurrent control clinical observation. SETTING: Diagnosis and Treatment Center for Coronary Heart Disease, the 305 Hospital of Chinese PLA. PARTICIPANTS: A total of 156 inpatients and/or outpatients, who were recently confirmed with primary hypertension, were recruited for this study between June 2005 and April 2007. Patients consisted of 114 males and 42 females. All patients averaged 56 ± 4 years of age (range: 40-75). Inclusive criteria: Corresponding to diagnostic standards for preventing and treating hypertension instituted in 2004 by Chinese scholars. Confirmed as primary hypertension within 1 month. Not receiving any blood pressure lowering, hypolipidemic or nitrate-like drug treatments. Written informed consents for laboratory measurements were obtained from all subjects. The study was approved by the hospital's Ethics Committee. METHODS: According to the mean pulse pressure over 24 hours, all patients were assigned into 3 groups: Group A (mean pulse pressure ＜ 40 mm Hg, n=92), group B (40 mm Hg ≤ mean pulse pressure ＜ 60 mm Hg, n=39) and group C (mean pulse pressure ＞ 60 mm Hg, n=25). In each group, daytime pulse pressure and night-time pulse pressure, as well as 24-hour mean pulse pressure were measured using a non-invasive portable ambulatory blood pressure monitor (ABPM-04, Meditech Inc, USA). Carotid-femoral and carotid-radial arterial pulse wave velocities were measured using an automatic pulse wave velocity determinator to evaluate large artery dilation. Blood flow mediated and nitroglycerin-dependent dilatation of the brachial artery was determined using a high-resolution ultrasound technique to evaluate vascular endothelial function. MAIN OUTCOME MEASURES: Correlations of ambulatory pulse pressure to large artery dilation and arterial endothelial function. RESULTS: All 156 patients were included in the final analysis. Correlation of ambulatory pulse pressure to large artery dilation: Carotid-femoral arterial pulse wave velocity was significantly positively correlated to daytime pulse pressure, night-time pulse pressure and 24-hour mean pulse pressure, with coefficient of partial correlation being 0.310, 0.281 and 0.303, respectively, P ＜ 0.01). There were no significant correlations of carotid-radial arterial pulse wave velocity to daytime pulse pressure, night-time pulse pressure or 24-hour pulse pressure (P ＞ 0.05). Correlation of ambulatory pulse pressure to arterial endothelial function: There was a linear relationship between ambulatory pulse pressure and blood flow-mediated blood vessel dilatation values. Linear correlation analysis was performed, taking ambulatory pulse pressure as an independent variable, and endothelial-dependent dilatation as a dependent variable. Results demonstrated that blood flow-mediated blood vessel dilatation was significantly negatively correlated to daytime pulse pressure, night-time pulse pressure and 24- hour mean pulse pressure (r = -0.684, -0.597, -0.668, P ＜ 0.01). There was no correlation of ambulatory pulse pressure to non-endothelial-dependent blood vessel dilatation. CONCLUSION: Ambulatory pulse pressure increase is closely related to large artery elasticity decrease and injury to endothelial function in patients with primary hypertension.

In vitro selection of specific aptamers against microcystin-LR / 中华预防医学杂志

<p><b>OBJECTIVE</b>In vitro selection of specific RNA aptamers against microcystin-LR from a random RNA pool.</p><p><b>METHODS</b>A RNA library with 40 randomized nucleotide positions was applied to select for specific aptamers to microcystin-LR covalently linked to Sepharose by using a standard in vitro selection protocol.</p><p><b>RESULTS</b>The specific enriched RNA aptamer for microcystin-LR increased step by step from initial round to 11th round after which a plateau of the aptamer quantity was observed between 11th and 13th round. The enriched RNAs from last round were reverse transcribed, PCR amplified and cloned into E. coli DH10 b competent cells. Sixty colonies were sequenced from which 38 sequences were aligned and classified into 3 families and 5 duplicates and no conserved sequences were found among them. Eight representative clones from the groups were selected for further binding experiments comparing with original pool RNA. Four clone RNAs were identified with relatively high affinity to microcystin-LR, of which MC25 clone RNA could combine with microcystin-LR as lower as 0.5 micromol/L.</p><p><b>CONCLUSION</b>Subpopulations of RNA molecules that bind specifically to microcystin-LR have been isolated from a population of random sequence RNA molecules, which might provide a new way for future application in environmental monitoring of microcystin.</p>

Preliminary study on effect of cardiac cycle and respiratory motion on Doppler flow spectrum of common femoral venous in healthy adults / 中华超声影像学杂志

Objective To evaluate the nature of Doppler flow spectrum of normal lower limb venous and to assess the influence of respiratory and cardiac cycle on it. Methods The right common femoral veins of 32 healthy adults were evaluated by pulsed wave Doppler with simultaneous electrocardiogram and respiratory tracing. Results In normal respiration,the Doppler spectrum of common femoral vein was divided into three types. Type C(“C” is for cardiac) displayed a wave rhythm which shared similarity with the heart beat and consists of atrial systolic (a),systolic (s) and diastolic (d) wave. Cardiac waveforms were modulated by respiratory motion: during expiration,the velocity of s,d wave gradually increased and during inspiration gradually decreased,and a wave was on the opposite. Type R(“R” is for respiration) displayed a waveform which is in harmony with respiratory signal basically. Type CR displayed a waveform which is an integration of type C and type R. Conclusions During quiet respiration,lower limb venous Doppler flow spectrum is influenced both by respiratory and cardiac cycle. The appreciation of this phenomenon would be instructive on analyzing the Doppler spectrum of lower limb venous in normal and pathological condition.

Influence of cardiac motion on the velocity-time integrals of Doppler flow spectra: in vitro model study / 中华超声影像学杂志

Objective To investigate the influence of simulated heart motion on the Doppler spectrum velocity- time integral (VTI) of simulated blood flow measurements through an in vitro model. Methods Using heart-motion simulator model TD-3 designed by ourselves to note the feature of Doppler spectrum of simulated heart and simulated blood flow which moved separately and synchronously.The affection of the simulated heart's motion on the VTI of the simulated blood flow and their quantitative relationship were observed.Results When the simulated heart and blood flow moved synchronously, the VTI of the combined motion was the algebra sum of their VTI when their motion independently. The velocity and frequency of Doppler spectrum of simulated heart were unchanged. Conclusions The motion of simulated heart has a great influence on the value of Doppler blood flow spectrum VTI and this effect should be considered when blood flow volume was measured using Doppler's methods.