Objective To screen out single chain variable fragment antibody (scFv)specific to vascular cell adhesion mole‐cule 1(Vcam‐1)from phage recombinant antibody library ,and to evaluate its activity and compare its activity with full‐length monoclonal antibody.Methods Amplification of Vcam‐1 was performed by PCR and Vcam‐1 gene plasmid was transferred into eukaryotic cells to express Vcam‐1 antigen protein.Immune cuvette was coated with purified Vcam‐1 antigen ,and the positive clones were screened out by 4 rounds of “adhesion‐elution‐proliferation” process with gradually increasing pressure.The posi‐tive clones were tested by ELISA method and high titer clones were chosen for gene sequencing.Then the high‐titer clones were transferred into E.coli ,and the clone with the highest expression was regarded as the final requisite one.Competent cells were infected by the final requisite clone and scFv was expressed.After purification ,the activity of scFv was tested by ELISA and its affinity was evaluated.Results Molecular weight of Vcam‐1 antigen protein was 85-90 kD.Positive clones were screened out by taking Vcam‐1 protein as the antigen ,and 9 high titer clones were obtained by single phage ELISA.Gene sequencing of these clones was carried out and 3 sequences were obtained ,1 of which got the highest expression.Molecular weight of the expressed scFv was about 30 kD.The scFv got high affinity to Vcam‐1 antigen according to ELISA ,in spite of its lower activity than full‐length monoclonal antibody.Conclusion scFv antibody specific to Vcam‐1 was successfully obtained from phage display librar‐y ,which laid the foundation of subsequent in vivo diagnosis and therapy.

Objective To speculate the precipitating factors,clinical features and histopathological changes in patients with granuloma annulare (GA).Methods A study was performed in 53 patients with GA . Results The incidence of female was more prevalent than male in our study. The exposure of sunlight was found more relatedly in GGA than LGA.The onset age of GGA was inclined to be elder and GGA seemed to last longer.The histopathological study showed that palisading granuloma pattern accounted for 71.7% in all patients,and scattered histiocytic infiltration accounted for 28.3%.Such as cryotherapy,topical steroids were used for LGA and systemic administration was an alternative to GGA. Conclusions Ultraviolet may be associated with the development of generalized GA. The most common pattern is the palisading granuloma though the histopathological changes were variable.Topical therapy is effective in LGA,while the systemic therapy is mainly used for GGA.

Objective To evaluate the clinical value of quantitative tissue velocity imaging (QTVI) in assessing left ventricular(LV) global and regional myocardial function in patients with mitral valve replacement(MVR).Methods Eighty patients having their implantations for more than six months were examined by echocardiography.QTVI-derived parameters such as peak systolic velocity(Sa,Sm) and early diastolic velocity(Ea,Em) of mitral annulus and LV wall were measured from the apical four-chamber,two-chamber and long axis corresponding myocardial segments in MVR groups decreased and LV ejection fraction but negative correlation between Ea' and isovolumic relaxation time(IVRT') in patients(P<0.01).Conclusions QTVI plays an important role in determining LV function of patients after MVR accurately.

BACKGROUND: The pathological characteristics of pulmonary interstitial fibrosis are the proliferation of a large number of fibroblasts and the increasing deposition of matrix collagen that takes the place of normal lung structure. Fluvastatin can inhibit the proliferation of fibroblasts and many other cells.OBJECTIVE: To investigate the effects of fluvastatin in inhibiting the proliferation of rat lung fibroblasts cultured in vitro and its influence on bleomycin-induced pulmonary fibrosis and ventilation function.DESIGN: A randomized controlled trial.SETTING: Department of Respiratory Diseases, Xijing Hospital, Fourth Military Medical University of Chinese PLA; Teaching and Research Section of Pathology, Department of Basic Medicine, Fourth Military Medical University of Chinese PLA; Research Institute ofOrthopedics, Xijing Hospital,Fourth Military Medical University of Chinese PLA.PARTICIPANTS: The study was conducted in the laboratory of Department of Respiratory Diseases, Xijing Hospital of Fourth Military Medical University of Chinese PLA from January to December 2001. Thirty-one healthy adult male Sprague-Dawley(SD) rats of grade Ⅰ were selected in this study.INTERVENTIONS: The fibroblasts derived from the lung normal of one rat were cultured in vitro in media containing fluvastatin. The effect of fluvastatin on the growth curve and the effect of its different concentrations(0, 1 × 10-7,1 ×10-6, 1 ×10-5, 1 ×10-4, 1 ×10 3and 1 ×10-2 mol/L, fluvastatin of 0 mol/L was taken as the blank control group) in inhibiting the cultured cells were observed with MTT colorimetry. The effect of fluvastatin on the division index of the fibroblasts was analyzed by direct cell counting Hydroxyproline colorimetry was used to detect the influence of fluvastatin on the collagen secretion in the media. The other 30 SD rats were divided into six groups: normal control group, bleomycin-induced group and fluvastatin-treated groups(TH 1,TE1, TH15 and TL15 groups) named according to the date of giving fluvastatin,i. e. the 1st day and the 15th day, after the rats were given bleomycin A5. All the rats were killed 28 days later. The number of fibroblasts, the thickness of alveolar wall and the area of mesenchyma in lung tissue were measured by HE staining. The extracellular matrix and collagen in lung tissue were observed by Masson and sirius red staining, and hydroxyproline in lung tissue homogenates was measured.MAIN OUTCOME MEASURES: Fibroblast growth curve and division index of rat lung, hydroxyproline in the media and lung tissue homogenates,number of fibroblasts and the thickness of alveolar wall, the area of mesenchyma, extracellular matrix and collagen contents in lung tissue.RESULTS: Fluvastatin could inhibit the proliferation of the rat lung fibroblasts cultured in vitro(t=4.20 to 17.52, P < 0.01), and its inhibitory effect was increased with the increased dose of fluvastatin, which showed a dose-dependent effect. The 1 × 10-4 mol/L fluvastatin could completely inhibit the proliferation of the cultured cells, and the A490 value from the 2nd day on the fibroblasts by MTT colorimetry was not insignificantly different from those on the 1st day( P > 0.05) . The division index of the fibroblasts and secretion of collagen were obviously decreased by fluvastatin( t = 8. 037,P <0.01; t =3.99 to 10. 84, P <0.05 or P <0.01). In vivo, the number of fibroblasts, the thickness of lung alveolar wall, the area of mesenchyma and the content of hydroxyproline in lung tissue were significantly higher in bleomycin group than in control group( t =4. 62 to 11.93, P < 0. 01), while those in the fluvastatin-treated groups were lower than those in bleomycin group in different degrees( t = 2.69 to 7.65, P < 0.05 to 0.01 ) . The distribution of extracellular matrix and types Ⅰ and Ⅲ collagen in lung tissue were obviously increased in bleomycin group as compared with that in control group, but decreased in different degrees in fluvastatin-treated groups.CONCLUSION: Fluvastatin can significantly inhibit the proliferation of rat lung fibroblasts in vitro, suggesting that it may be an effective drug for pulmonary fibrosis. Treatment at earlier stage is more effective than at advanced stage.

Objective To investigate the imaging performance and feasibility of 99Tcm labeled scFv against VCAM-1(99Tcm-scFv-VCAM-1) on atherosclerosis model rabbits.Methods HYNIC was used as a chelator for 99Tcm labeling.The labeling efficiency and radiochemical purity of 99Tcm-scFv-VCAM-1 were measured by instant thin layer chromatography after PD-10 purification.New Zealand white rabbits were employed for establishing atherosclerotic animal models by endothelia immunity injury and high fat diet, and plaques at aorta lesions were examined by HE staining.Model rabbits were sacrificed after administration of 99Tcm-scFv-VCAM-1 at 1 or 2 h respectively, and tissue samples were measured with gamma counter and weighted to obtain in vivo biodistribution data.Planar imaging was performed 1 and 2 h after the injection of 99Tcm-scFv-VCAM-1 to investigate radioactivity of abdominal aorta.After imaging study, atherosclerosis plaque and VCAM-1 expression at aortas were confirmed by the immunohistochemistry (IHC) study.Two-sample t test was used to analyze data.Results 99Tcm-scFv-VCAM-1 was successfully synthesized.Its labeling efficiency was 75%-83%, radiochemistry purity was (98.54±1.03)% and specific activity was 216 MBq/nmol.Atherosclerosis plaque was confirmed at the aortas of experimental rabbits by HE staining, while no plaque was observed in controls.Biodistribution data indicated that the tracer was cleared mainly through the kidneys.Planar imaging showed that the tracer uptake in abdominal aorta of model rabbits was higher than that of control rabbits, the T/B ratios at 2 h of the model group and control group were statistically different (3.68±0.73 vs 2.42±0.39;t=2.950, P<0.05;n=5).Atherosclerosis plaque and high level of VCAM-1 expression were observed at aortas of model rabbits by IHC study.Conclusions It is feasible and effective to detect vulnerable plaques using 99Tcm-scFv-VCAM-1.It may provide a promising way for early diagnosis and accurate evaluation of atherosclerosis.

Metabolic syndrome (MS) is a group of syndromes caused by the disorder of metabolism of various substances in the body. The main clinical manifestations are dyslipidemia, central obesity, hypertension, abnormal glucose tolerance and insulin resistance. With the changes of diet structure and habits, the incidence rate of MS is increasing, and the patients are younger. It is an important factor in many diseases, such as diabetes, atherosclerosis, coronary heart disease, hyperlipidemia, cirrhosis and some cancers. MS has seriously affected people's lives and health. Central obesity and insulin resistance are recognized as important pathogenic factors. At present, the pathogenesis of MS and its components has not been fully understood. The clinical manifestations of metabolic syndrome are complex and diverse. Traditional Chinese medicine (TCM) believes that the occurrence of metabolic syndrome is related to such factors as proper diet, emotional disorders, excessive escape and little movement, old age and physical deficiency. TCM scholars have studied the pathogenesis of MS in such pathological factors as phlegm and blood stasis, such visceral functions as liver, spleen and kidney, roles of Qi and blood, and emotional factors. As the basic substance of organism, Qi is closely related to the process of metabolism. The occurrence of MS is closely related to the rise and fall of Qi moving to and from the body as well as the abnormal gasification function of the transformation of Qi. Qi is derived from the five internal organs, which are respectively called Heart Qi, liver Qi, spleen Qi, lung Qi and kidney Qi. The "Qi of the five internal organs" is involved in the whole process of the generation, transportation and excretion of the essence of the body. Based on the "Qi of five internal organs", this paper discusses the pathogenesis of MS with phlegm, blood stasis and water drink as pathological factors.

OBJECTIVETo establish an HPLC-ELSD internal standard method for determination of astragaloside IV in Astragali Radix.

METHODWith Ginsenoside Rb2 as internal standard, the separation were carried out on an Agilent TC-C18 (4.6 mm x 150 mm, 3.5 microm) column with methanol-water (72: 28) as mobile phase. The flow rate was 1.0 mL x min(-1) and the drift tube temperature of the ELSD was 75 degrees C. The gas pressure was set at 172.4 kPa using the clean and dry compressed air as spray gas.

RESULTThere was good linearity in the range of 0.5624-5.624 microg of astragaloside IV (r = 0.9999); The average recovery was 98.06% with RSD of 0. 98%.

CONCLUSIONThe internal standard method is accurate and reproducible, and suitable for quality control of radix astragali.

Astragalus Plant ; chemistry ; Chromatography, High Pressure Liquid ; standards ; Drugs, Chinese Herbal ; analysis ; Linear Models ; Reference Standards ; Saponins ; analysis ; Triterpenes ; analysis

Objective:To investigate the effect of nonalcoholic fatty liver disease (NAFLD) on glucose and lipid metabolism during pregnancy.Methods:A retrospective analysis was performed in women who gave birth in Minhang Hospital Affiliated to Fudan University from January 2013 to June 2020. The data on demographic, clinical examination, and delivery were obtained via electronic medical record abstraction. According to the ultrasound imaging, all pregnant women were divided into NAFLD group and control group. The difference of glucose and lipid metabolism indexes, incidence of gestational diabetes mellitus, and gestational hypertension between two groups were compared. Logistic regression model was used to examine potential associations between NAFLD and metabolic related adverse pregnancy outcomes.Results:A total of 14 708 pregnant women with a mean age of (29.1±4.7) years and a mean body mass index of (21.0±2.8) kg/m 2 were included in our study. Of those eligible women, 554 (3.8%) were confirmed by ultrasound as NAFLD. Pregnant women with NAFLD presented higher circulating levels of fasting glucose [(4.2±0.5)mmol/L vs (4.1±0.5)mmol/L, P<0.01], 1 h plasma glucose [(7.4±1.7)mmol/L vs (6.6±1.6)mmol/L, P<0.01] and 2 h plasma glucose [(6.2±1.4)mmol/L vs (5.7±1.3)mmol/L, P<0.01] after glucose loading, HbA 1C [(5.2±0.4)% vs (5.1±0.5)%, P<0.01], triglyceride [(2.1±1.1)mmol/L vs (1.6±0.7)mmol/L, P<0.01], total cholesterol [(4.8±0.8)mmol/L vs (4.7±0.9)mmol/L, P<0.01], low density lipoprotein-cholesterol [(2.6±0.7)mmol/L vs (2.5±0.7)mmol/L, P<0.01], uric acid [(224.1±51.8)μmol/L vs (203.0±45.9)μmol/L, P<0.01] level. After adjusting for potential confounders, NAFLD significantly increased the risk of gestational diabetes mellitus ( OR=1.722, 95% CI 1.079-2.747, P=0.023) and gestational hypertension ( OR=3.845, 95% CI 2.247-6.582, P<0.001). Conclusions:Compared to non NAFLD, women with a diagnosis of NAFLD had more significant glucose and lipid metablic aberrations during pregnancy and increased incidence of gestational diabetes and gestational hypertension. Pregnant women with NAFLD should be closely monitored on glucose and lipid metabolism and blood pressure to prevent gestational diabetes mellitus and hypertension.

Objective: To investigate the relationship between fine motor skills and executive function in school aged boys with attention deficit hyperactivity disorder (ADHD) aged 6-10 years, so as to provide a reference for promoting executive function. Methods: From November 2022 to May 2021,65 boys with ADHD were recruited from a tertiary hospital in Beijing and a public elementary school in Beijing, and 65 boys with typical development according to age and sex were recruited as controls. Children s fine motor skills were assessed using the movement assessment battery for children, second version (MABC-2), and children s inhibitory control, working memory, and cognitive flexibility were assessed using the Stroop color word test (SCWT), Rey osterrich complex figure test (ROCFT) and trail making test (TMT), respectively. Independent samples t-tests were used to analyze the differences between the two groups, and Pearson correlation analysis and stratified regression analysis were used to explore the relationship between fine motor skills and executive functioning in boys with ADHD. Results: In the fine motor skills test, boys with ADHD scored significantly lower (8.68±3.62) than control boys (11.22±2.27)( t =4.80, P <0.05); in the executive function test, the results of SCWT, ROCFT, and TMT tests were significantly worse in boys with ADHD than in normal boys ( t =-4.53-4.42, P < 0.05). Correlation analysis showed that the fine motor skill scores of boys with ADHD were negatively correlated with the number of word sense errors ( r =-0.35) and color errors ( r =-0.42), and positively correlated with memory scores for delayed structure ( r =0.30) and detail ( r =0.25), which were negatively correlated with TMT-A ( r =-0.34),TMT-B reaction time ( r =-0.26), number of errors ( r =-0.43) ( P <0.05). Stratified regression analysis showed that fine motor skill scores of boys with ADHD were predictive of test results for inhibitory control, working memory and cognitive flexibility, with explanatory rates of 15%, 7%, and 19%, respectively. Conclusion Fine motor skills in boys with ADHD are correlated with executive functions, and fine motor skills could predict inhibitory control, working memory and cognitive flexibility to some extent. The development of fine motor skills in boy with ADHD could promote their executive functions.

Objective:To observe any effect of electroacupuncture applied to the Jialianquan points in treating post-stroke oropharyngeal dysphagia.Methods:Forty-five stroke survivors with oropharyngeal dysphagia were randomly divided into an electroacupuncture group ( n=15), a neuromuscular electrical stimulation group ( n=15) and a control group ( n=15). All groups received 30 minutes of routine swallowing function training 5 times a week for 3 weeks. The electroacupuncture group was additionally provided with 30 minutes of electroacupuncture applied to the Jialianquan (CV23) points, and the neuromuscular electrical stimulation group instead received 30 minutes of neuromuscular electrical stimulation over the bilateral submental muscles. Videofluoroscopic swallowing studies were performed before and after the 3 weeks of treatment. Standardized swallowing assessment was conducted producing functional oral intake scale ratings, modified barium swallow impairment profiles and the penetration-aspiration scale scores. Surface electromyography was also employed to evaluate submental muscle functioning through measuring the swallowing time, average EMG (AEMG) value and peak amplitude. Results:After the treatment, significant improvement was observed in all of the evaluations with both groups, but the average scores were significantly better in the electroacupuncture and neuromuscular electrical stimulation groups compared with the control group and significantly better in the electroacupuncture group than in the neuromuscular electrical stimulation group.Conclusion:Electroacupuncture at the Jialianquan point can significantly improve the swallowing of stroke survivors with oropharyngeal dysphagia. It is more effective than neuromuscular electric stimulation.