Preparation of anti-GPC3 single chain antibody for targeted detection of hepatocellular carcinoma / 生物工程学报

Glypican-3 (GPC3) is a key member of Glypican family and plays an important role in the development, angiogenesis and metastasis of hepatocellular carcinoma (HCC). Most HCC overexpresses GPC3, but GPC3 is hardly detected in normal adult liver and benign liver lesions, so it is regarded as a highly specific diagnostic marker and an ideal therapeutic target for HCC. In this study, we cloned the heavy and light chain variable region gene from the monoclonal antibody targeted to GPC3 screened in the previous stage, and linked it with a segment of flexible peptide (Linker) to obtain the single chain antibody against GPC3. The single chain antibody gene was cloned into vector for prokaryotic expression and purified to obtain high purity protein. Detection shows that the single-chain antibody produced by us has the same binding activity with the full-length antibody, and can accurately target the tumor site of Huh7 tumor-bearing model mice after coupling Cy5.5 fluorescence, suggesting that the single-chain antibody has the potential to realize multi-directional liver cancer precise surgical navigation under the guidance of a probe.

An Insulin Molecularly Imprinted Electrochemical Sensor Based on Epitope Imprinting / 分析化学

A novel molecularly imprinted electrochemical sensor for direct detection of insulin was prepared based on epitope imprinting.C-Terminal polypeptide in insulin as template molecule was firstly self-assembled on the Au electrode.Then the molecularly imprinted polymer (MIP) was fabricated by electropolymerization with o-phenylenediamine (o-PD) as functional monomer on this Au electrode.After elution of template molecules by NaOH solution, the imprinting cavities were formed with the three-dimensional structure matched with the polypeptide in insulin molecules.The imprinting cavities could specifically recognize and rebind with insulin molecules.With K3[Fe(CN)6]/K4[Fe(CN)6] as a probe, the insulin was indirectly detected.There was a linear relationship between the response current and the insulin concentrations in the range of 1.0 × 10-14-5.0 × 10-13 mol/L, and the detection limit was 7.24×10-15 mol/L.The developed sensor exhibited good selectivity and stability, and could be applied to the determination of serum samples.

Role of mimecan in development of atherosclerosis induced by increased blood pressure variability / 中南大学学报(医学版)

Objective:To examine the changes of mimecan protein expression in development of atherosclerosis induced by sinoaortic denervation,and to explore the effects of mimecan knock down on the proliferation and migration of vascular smooth muscle cells.Methods:The animals were randomly divided into a sham group and a model group (n=8 in each group).The rat model of blood pressure variability was established by sinoaortic denervation,and the hemodynamic indexes were recorded 20 weeks after the surgery to confirm the success of the model.The thoracic aorta was excised and stained with immunohistochemistry to observe the pathological changes of smooth muscle tissues and the changes of mimecan expression.The mice vascular smooth muscle cells were isolated,and which were treated with mimecan siRNA to knock down the mimecan expression,The cell proliferation was observed by 5-ethynyl-2'-deoxyuridine (Edu) in corporation test and the changes of cell migration was observed by wound healing test.Results:Twenty weeks after sinoaortic denervation,the blood pressure variability in the model group was significantly increased compared with that in the sham group,suggesting the model was successfully established.In addition,the increased blood pressure variability in the model group promoted the proliferation and migration of the vascular smooth muscle cells in thoracic aorta,while the expression of mimecan protein was significantly decreased.In in vitro assays,the knock down of mimecan in mice vascular smooth muscle cells could promote the cell proliferation and migration.Conclusion:Mimecan plays a protective role in the development of sinoaortic denervation induced atherosclerosis through amechanism involving suppression of the proliferation and migration of vascular smooth muscle cells.

Effect of oxysophoridine on electric activities and its power spectrum of reticular formation in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of oxysophoridine (OSR) on the EEG and its power spectrum of reticulum formation in mesencephalon of anaesthetized rat.</p><p><b>METHOD</b>Utilizing the technique of brain stereotactic apparatus, electrodes were implanted into reticulum formation of mesencephalon. Monopolar lead and computerized FFT technique were employed to record and analyse the index of EEG, power spectrum and frequency distribution in order to study the effect of oxysophoridine on the bioelectricity change of mesencephalon reticulum formation in rats.</p><p><b>RESULT</b>After administrating(icy) with oxysophoridine at the dose of 2.5,5, 10 mg/rat, the EEG of mesencephalon reticulum formation mainly characterized with low amplitude and slow waves accompanied by spindle-formed sleeping waves with a significant decrease of total power of EEG (P < 0.05) while the ratio of theta, alpha waves increased in total frequency of rats (P < 0.05).</p><p><b>CONCLUSION</b>Oxysophoridine possesses central inhibitory effects and its inhibitory mechanism may associate with the reduction of bioelectricity in mesencephalon reticulum formation. Mesencephalon reticulum formation may serve as one part of the structure serving as the circuit conducting the central inhibitory effect of oxysophoridine. [Key words] oxysophoridine; reticulum formation; electroencephalogram (EEG) ; rats</p>

Coronary characteristics of acute coronary syndrome with ischemic J wave / 中国综合临床

Objective To understand the coronary characteristic of acute coronary syndrome patients with ischemic J wave. Methods Comparison was made between 60 acute coronary syndrome patients with ischemic J wave and 60 acute coronary syndrome patients without ischemic J wave. All patients were examined by Holter monitoring electrocardiogram and coronary arteriongraphy. To distinguish the stenosis degree was defined in three degree based on Genisini score of 0, 1-80, and 81-160. Plaque types were divided into Ⅰ,Ⅱ, Ⅲ by Ambrose classification,the coronary and plaque characteristics of acute coronary patients were observed with ischemic J wave. Results There were no significant difference of stenosis degree between the two groups ( U = 3. 0686, P = 0. 0022), whereas there were significant difference of plaque types (x2 =16. 0890, P = 0. 0003) and coronary vessel numbers(x2 =12. 1045, P = 0. 0024). The degree of stenosis, the plaque types, and number of stent in coronary vessel were positively correlated with ischemic J wave(r =0. 44,0. 34,0. 31 ;P <0. 05). Conclusions The acute coronary syndrome with ischemic J wave patients is often not only accompanied with serious coronary stenosis and high incidence rate of multivessel disease,but also high probability of unstabilized plaque. Ischemic J wave can be a predictor of super-acute ischemic of myocardium.

Stabilometer test for the quantified evaluation of equilibrium function in stroke patients with hemiplegia / 中国组织工程研究

BACKGROUND: The patients with hemiplegia of stroke commonly appear balance disturbance and the mean of assessment on it is applied in quality commonly. But the implementation of assessment on rehabilitation schedule and therapeutic results requires much more quantitative standards on balance function.OBJECTIVE: To probe into interrelationship between every parameter of stabilometer test and variables in other function evaluations in the patients with hemiplegia of stroke as well as the value of evaluation on functional recovery in the patients with hemiplegia of stroke.DESIGN: Quantitative assessment based on the qualitative assessment.SETTING: Department of Otolaryngology and Department of Rehabilitation Medicine in General Hospital of Chinese PLA.PARTICIPANTS: Nineteen first-attack cases of hemiplegia of stroke received rehabilitation treatment in Department of Rehabilitation Medicine of General Hospital of Chinese PLA from September 2002 to April 2003, diagnosed as disorder of cerebral hemisphere on lateral internal carotid artery. Of those, 14 cases were males, 5 cases females; aged varied from 33 to 55 years and the course of sickness was at the range from 0.5 to 4 months.METHODS: Nineteen patients with hemiplegia of stroke received functional independence measures(FIM), timed up-go test(TUGT), Berg balance scale (BBS) and evaluation on balance measure with stabilometer. Of those, stabilometer is a kind of quantitative analysis method, including the parameters of total length of locus, speed, peripheral area, average central changing site of X wavering direction, average central changing site of Y wavering direction, etc.MAIN OUTCOME MEASURES: Pearson correlation analysis was used to determine the interrelationship between every parameter measured with stabilometer and other measured functional variables.RESULTS: By intention-to-treat management analysis, 19 cases of hemiplegia of stroke all entered analysis on results. Among the parameters of stabilometer test, the moderate negative correlation presented in the comparison of total length of locus, speed and peripheral area with BBS during eyes opened and closed ( r = - 0. 475 to - 0. 705, P ＜ 0.05), of which, the correlation between peripheral area and BBS score during eyes closed was better(r= -0.475, P ＜ 0. 05) . Correlation did not present in the comparison of average central changing site of X wavering direction and average central changing site of Y wavering direction with BBS during eyes opened and closed( r = - 0. 346 to 0. 234, P ＞ 0.05) . Moderate positive correlation presented in the comparison of TUGT result with total length of locus, speed and peripheral area during eyes closed( r = 0. 508 to 0. 583, P ＜ 0.05). The correlation did not present in the comparison of the parameters determined with stabilometer and FIM ( r = - 0. 048 to - 0. 296, P ＞ 0.05).CONCLUSION: Stabilometer test can determine balance function in quantity in patients with hemiplegia of stroke, and reflect the severity of balance disturbance, of which, total length of locus speed and peripheral area during eyes opened and closed can judge balance function of patients sensibly and reliably.

Rotation test versus water caloric test in the evaluation of the vestibular function / 中国组织工程研究

BACKGROUND: There are two methods to evaluate the vestibular function, the rotation test and the water caloric test. What is the relativity between the test results with these two methods?OBJECTIVE: To observe the consistency between the vestibular functional evaluations with the rotation test and the water caloric test.DESIGN: A comparative study on the advantage of the evaluation methodologies with patients as the objects.SETTING: Department of Otolaryngology of Chinese PLA General Hospital.PARTICIPANTS: The patients with vertigo, deafness and tinnitus were chosen, who were picked up in the inpatient department and outpatient department in the Otolaryngology Department of the Chinese PLA General Hospital from April 1990 to December 1996. With the permission, the vestibular functions of all the patients were tested to be abnormal by the Hallpike caloric test. There were 68 males and 32 females, whose ages varied from 16 to 74, and the average age was 37 ± 3. There were 91 cases with unilateral abnormality and 9 cases with bilateral abnormally. The judging standard of the abnormal vestibular function is that slow phase velocity (SPV) of the nystagmus on the normal side-SPV on the abnormal side /SPV on the normal side-SPV on the abnormal side × 100% ] ≥ 15%, and the ratio ＜ 15% means normal.METHODS: The rotation test was done to the patients with the sine swing stimulation and rotation sudden pause stimulation respectively . The stimulation scope of the sine swing stimulation was 90° and the frequency was 0. 167 Hz; In the rotation sudden pause test, when the peak velocity reached 90°/s at the acceleration of 1°- 2°/s2, the velocity was maintained until the disappearance of the nystagmus, then sudden pause was done for the further test.MAIN OUTCOME MEASURES: The results of the sine swing test and the rotation sudden pause test.RESULTS: According to the actual disposal, in the rotation test, there were 9 cases with bilateral abnormality, among them the nystagmus was induced in 8 cases and no nystagmus was induced in 1 case with bilateral acoustic neuroma, and in the other cases the nystagmus was induced. In the water caloric test, there were 91 cases with unilateral abnormality, 2 cases refused to take the rotation sudden pause test. In the sine swing test, 64% (58/91)cases had abnormal vestibular function, and in the rotation sudden pause test 61% (54/91) cases had abnormal vestibular function.CONCLUSION: The abnormality of the vestibular function can be found in the sine swing test and the rotation sudden pause test of the rotation test, but the results do not completely conform to that in the water caloric test.

Quantitative evaluation of vestibular function, balance function and optokinetic function in elderly person / 中国组织工程研究

BACKGROUND: Old people above 80 years old have a high incidence of dizziness and tumble, which are associated with weakening vestibular function, balance function and optokinetic function.OBJECTIVE:This study was designed to make a quantitative evaluation of vestibular function,balance function and optokinetic function in people over 80 years old.DESIGN:It was a quantitative study on posture function(vestibular and balance function),auditory function(frequency audiometry)and visual function (optokinetic function) in elderly persons.SETTING:Department of Otolaryngology ,General Hospital of Chinese PLA.PARTICIPANTS:Totally 41 old male inpatients with systemic diseases in Rehabilitation Department of the General Hospital of Chinese PLA, with an age from 80 to 94 were selected from March 1993 to January 1994.Of them,12 had tinnitus,20 had dizziness.During the same period,old cadres including 11 males and 7 females aged from 60 to 79 years with normal findings of physical examinations were selected as controls. No participant in this group suffered from tinnitus, deafness and dizziness. All the participants were voluntary.METHODS: Patients underwent full-scale otolaryngological examinations.Audiometry consisted of battery of tests including frequency audiometry and evoked auditory brainstem responses(ABR). Vestibular function (posture) was assessed by vestibular-ocular reflexes. Participants accelerated from 1 /s to 90 /s. Then, the angular speed was kept until the nystagmus disappeared,at the time a sudden stop was made.Changes in nystagmus during the tests were recorded.The eyes firstly turned to left and 10 minutes later turned to right.Slow-phase angular velocity was recorded and the imbalanced left/right rate was calculated.Vestibularspinal reflex (VSR) was measured using balance beam and the length of the sway of the center of gravity (CG) and the sway velocity of CG were all calculated. Balance tests using a cursor were employed to examine the central optokinetic system, including sweeping, tracking, and optokinetic responses.MAIN OUTCOME MEASURES: Frequency audiometry, vestibular-ocular reflexes, and vestibular-spinal reflex and optokinetic responses were examined to assess the posture function, auditory function and visual function in elderly persons over 80 years old.And the results were compared with those of balance function tests and visual function tests in old population aged from 60 to 79 years old.RESULTS:Practically,the number of old participants over 80 years old receiving the tests of frequency audiometry, vestibular-ocular reflexes, and vestibular-spinal reflex and optokinetic responses were 41,24,39,40 and 34 respectively;The number of 60-79 participants receiving balance tests and visual tests were 18 and 10.Individuals refusing to participate were omitted. ① Auditory function:In 41 elderly persons ＞80 years,13 ears (16％) had a normal middle frequency audiometry, 69 ears (84％) had a loss of hearing. ② Posture function: 15 (62％) had normal vestibular-ocular reflexes, and 9 (38％) had abnormalities in vestibular-ocular reflexes.Ten (26％) had a normal vestibular-spinal reflex and 29 (74％) had an abnormal vestibular-spinal reflex.③ Balance function:Of all the variables of balance function,the length of the sway of CG and the sway velocity of CG were most valuable. In populations in both brackets, the length of the sway of CG and the sway velocity of CG in eyes-closed condition were longer and faster than those in eyes-opened .condition (P ＜ 0.01). In over 80 years group,the length of the sway of CG and the sway velocity of CG were longer and faster than in 60-79 group (P ＜ 0.01). ④ Optokinetic responses:Optokinetic plus were not obviously different between over-80 group and 60-79 group (P ＞ 0.05) and the latent period of sweeping and the distortion of tracking were significantly longer and greater in former group than in later(P ＜ 0.05-0.01 ).CONCLUSION:Most of the elderly persons over 80 years have a decrease in auditory and posture functions.Their optokinetic and balance functions are degenerated more severely than 60-79 years old population.

Consistency of three measures commonly used in the assessment of balance and functional outcome of patients with post-stroke hemiplegia / 中国组织工程研究

BACKGROUND: To assess the balance function of post-stoke patients would be conducive to establish rehabilitative goal and to set up therapeutic plan.OBJECTIVE: To investigate the relationship between parameters of three commonly used measures and those of other functional evaluation for patients with post-stroke hemiplegia.DESIGN: Analysis of the relationship between parameters of balance rating scales.SETTING: At otolaryngology department and rehabilitative medicine department of the general military hospital.PARTICIPANTS: From September 2002 to July 2003, totally 57 patients with post-stroke hemiplegia, 45 males and 12 females, aged from 14 to 78years old, with the course of disease ranged from 0. 3 to 11 months, received rehabilitative treatment at rehabilitative department of the general military hospital, were selected to take part in the study. All were confirmed of internal carotid artery system lateral hemispheric disease.METHODS: Three balance measures, namely Berg balance scale, modified Balance subscale of the Fugl-Meyer test and Postural assessment scale for stroke patients were administered on 57 patients for evaluating their lower limb motor function, daily life capability and walking ability after rehabilitative treatments. The relationship between three measures was analyzed with Pearson-correlative analysis, and the relationship between three measures and lower limb motor function, daily life capability and walking ability were also investigated.MAIN OUTCOME MEASURES: ① relationship between three measures;② relationship between three measures and lower limb motor function, daily life capability and walking ability.RESULTS: The data from 57 stroke hemiplegic patients was analyzed without missing. ① Scores for Berg balance assessment, Fugl-Meyer balance rating scale and posture controlling scale for stroke patients was 46.70 ± 5. 15, 10. 26 ± 1.58 and 32.56 ± 2.06, respectively, and the scores of lower limb motor function in Fugl-Meyer assessment was 22. 35 ± 5.82,and moving and marching item in functional independence assessment 24.95 ± 5.2, 10 m maximum walking speed(49.06 ±24.68) m/s. The three measures were proved of significantly higher consistency( r = 0.613-0. 766,P ＜ 0.001 ). ② The three measures were also found to be obviously associated with the lower limb motor function, daily life capability and walking ability of patients with stroke( r = 0. 479-0. 718, P ＜ 0. 001 ).CONCLUSION: Berg balance assessment, modified Fugl-Meyer balance rating scale and posture-controlling scale have been proved of higher consistency and efficacy for assessing the balance function of post-stroke patients, thereby can be used as predictor for the balance functional outcomes,as well as the recovery of the lower limb motor function, daily life capability and walking ability of post-stroke patients.

Changes in Ca2+-ATPase in a guinea pig endolymphatic hydrops model / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the localization of Ca(2+)-ATPase (Ca(2+) pump) in the cochlear and its change after endolymphatic hydrops, and to study the relationship between compound action potential (CAP) threshold and the Ca(2+)-ATPase activety.</p><p><b>METHODS</b>The left endolymphatic sac was ablated to induce endolymphatic hydrops in fourteen healthy guinea pigs with normal action potential thresholds measured after a sliver ball electrode placed on the round window. Ca(2+)-ATPase activity was studied cytochemically using a lead citrate reaction in control and hydropic ears. The reaction product was lead phosphate particles as an expression of Ca(2+)-ATPase activity, observed with an eletron microscope.</p><p><b>RESULTS</b>Ca(2+)-ATPase activity is mainly found on the endolymphatic surface of Reis sner's membrane, the stereocilia and cuticular plate of inner and outer hair cells, and along the infolded plasma membrane of strial marginal cells. CAP thresholds of filtered click are increased and Ca(2+)-ATPase activity significantly decreased after endolymphatic hydrops in the mentioned locations.</p><p><b>CONCLUSIONS</b>CAP thresholds are increased and Ca(2+)-ATPase activity are significantly decreased in the cochlea after endolymphatic hydrops. These results suggest that there is a negative correlation between them.</p>

THE EFFECT OF QUANTUM DOTS ON CYTOCHEMISTRY OF MICE ABDOMINAL CAVITY MACROPHAGES IN VITRO / 解剖学报

Objective Study the effect of quantum dots(QDs) on macrophage cytochemistry and enzyme activity of mice in vitro. Methods To observe the macrophage biocompatibility with QDs and effect of QDs on PAS reaction,Feulgen reaction,ATPase,acid phosphatase(AcP),alkaline phosphatase(ALP),ANAE,SDH and LDH using inverted phase contrast microscope,fluorescence microscope,and cytochemistry methods were used.Results QDs hadn't effect to macrophage structure in(3.125mg/L) density of QDs,but its cytochemistry and cell enzyme activity had changed differently,that was PAS reaction,Feulgen reaction,AcP,ALP,ANAE and Mg~(2+)-ATP showed positive,and SDH and LDH showed negative.Feulgen reaction,ANAE,ALP and Mg~(2+)-ATP had statistical significance with the control group(P0.05).Conclusion In cell level,macrophage enzymes have changed differently by quantum dots,but QDs don't affect macrophage structure and phagocytosis. QDs can be applied to label living cell in invariably limited dose of QDs in biomedicine area,and cells doesn't have visible influence.

THE LOCALIZATION AND EFFECT OF QUANTUM DOTS ON ULTRASTRUCTURE OF MOUSE ABDOMINAL CAVITY MACROPHAGES IN VITRO / 解剖学报

Objective To observe the distribution and the effect of the quantum dots(QDs) on mouse abdominal cavity macrophages.Methods The QDs were co-cultured with mouse abdominal cavity macrophages in vitro.The differentiation and effect of the QDs on macrophage ultrastructures were observed under electronic microscope. Results The QDs were enveloped with unit membrane and internalized in the cytoplasm of the macrophage under transmission electron microscope.And it formed vacuolelike structures in the macrophage.There were many lamellar processes on the surface of the macrophage under scanning electron microscope.Conclusion The QDs can promote macrophage activation,and make its surface projection increased.The QDs were internalized by the macrophage,distributed in the cytoplasm,and formed vacuolelike structures enveloped with unit membrane.