3.The pleural membrane biopsy accumulates the fluid patient's diagnosis value to the chest cavity
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To appraise the diagnosis value of pleural membrane biopsy on pleural effussion.Meth- ods 218 patients with pleural effusion who were had pleual menbrane biopsy were retrospectively analyzed.Among them 165 cases were done only once,42 cases were done twice,11 cases were done three times.They were done 271 times of biopsies.Results The pleural membrane biopsy always masculine gender rate was 72.0%,1 pleural mem- brane biopsy succeeds the masculine gender rate was 59.6%,after the masculine gender rate which 2 pleural mem- branes biopsies succeeds was 69.7%.The illness complication formation rate was 8.7%.Conclusion Pleural mem- brane biopsy is a simply,safe and higher diagnosing-rate method.The different spot pleural membrane biopsy may cnhance diagnosis rate.
4.Affinity maturation of human scFv anti-TSLP in vitro
Dequn XIAN ; Siji NIAN ; Yingchun YE ; Wenfeng XU ; Qing YUAN
Chinese Journal of Immunology 2017;33(9):1301-1305
Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.
5.Selection and identification of full human scFv against TSLP
Jianguang ZHU ; Qing YUAN ; Li HUANG ; Wenfeng XU ; Siji NIAN
Chinese Journal of Immunology 2014;(12):1662-1665,1669
Objective:Expression of protein TSLP and selection of full human anti-TSLP single chain Fv ( scFv).Methods:The cDNA of TSLP was amplified.The amplified target gene and the expression vector pET 101/D-TOPO were ligated , and then transformed into E.coli BL21.The protein was induced to expression by IPTG and purified and identified.The biotinylated TSLP protein was used as antigen to select of human TSLP scFv from a constructed human scFv library by phage display .Results: The size of amplified cDNA of TSLP was about 423 bp,and that of expressed protein was about 26 kD.Dot blot and Western blot results showed that the expressed protein was correct.The constructed human scFv library was enriched for three rounds using biotinylated TSLP as antigen by phage display.ELISA results showed that 35% scFvs had binding activity with TSLP.The scFvs with good binding activity were selected and identified by Western blot and sequencing.Conclusion: The full human scFvs against for TSLP were selected suc-cessfully.
6.Clone and soluble fusion expression ofα-HL of Staphylococcus aureus
Tong WU ; Wenfeng XU ; Siji NIAN ; Yingchun YE ; Qing YUAN
Chinese Journal of Immunology 2016;32(4):532-535,541
Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.
7.Survey of precipitants of epileptic seizure
Yanxiao YIN ; Nian YU ; Longxiang ZHOU ; Yongfei CHENG ; Qing DI
Chinese Journal of Neurology 2017;50(4):255-260,282
Objective To investigate precipitants of epileptic seizure, and to explore the correlation between various precipitants and relationship between precipitants and clinical features of epilepsy.Methods Data were collected from 154 patients attending a tertiary-care epilepsy clinic of Nanjing Brain Hospital between April 2015 and April 2016.The patients with epilepsy were older than 16 years, had a clinical history of one year or more, and one seizure at least a year and one seizure at least in the latest three months.An enclosed questionnaire was combined with open interview to identify and characterize seizure precipitants and clinical characteristics of patients.Patients were asked respectively whether there were some precipitants three months before and during last three months.Correlation between seizure precipitants and relationship between precipitants and clinical characteristics, such as age, gender, course, seizure frequency and so on, were calculated.Results A total of 125 (81.2%) participants reported at least one precipitant.Common precipitants (in descending order) were as follows: emotional stress (56.0%), sleep disorder (38.4%), fatigue (27.2%), missed medication (20.0%).There were one to six different precipitants for one patient, and 60.8% of patients had two or more precipitants.There was a correlation between emotional stress and sleep disorders as well as fatigue (χ2=4.665, 8.668;P<0.05).Patients with idiopathic epilepsy were more sensitive to sleep disorders.There was no relationship between total precipitants and clinical features such as age, gender, age of onset, duration, type of seizure, seizure frequency, number of drug taking and so on.Conclusions Seizure precipitants were found widespread.The most common precipitants were found to be emotional stress, sleep disorders, fatigue and missed medication.There existed a correlation between emotional stress and sleep disorders as well as fatigue.There was no connection between total precipitants and patient′s demographic characteristics as well as clinical features.However, the type of seizure precipitants was different in patients with different demographic and clinical characteristics.
8.Sociodemographic and clinical factors associated with interictal depression symptoms in adults with epilepsy
Yong HU ; Qing DI ; Nian YU ; Yanfang ZHANG ; Lingying SU
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(3):241-243
Objective To investigate the prevalence of adult epileptic patients with interictal depression symptoms(IDs) and identify early predictors of IDs. Methods Adult patients with epilepsy were recruited ( n =110,45 females and 65 males) ,age between 16 and 67 years ( median 24 years). The sociodemographic and clinical factors of patients were recorded. Hamilton Depression Scale ( HAMD ) were applied to evaluate interictal symptoms of depression ( at least 72 hours after the last epileptic seizure). According to HAMD score,the epileptic patients were divided into IDs ( ≥8 ) and non-IDs(<8) groups. The sociodemographic and clinical factors were compared between the two groups to identify the prevalence and early predictors of IDs in adult epileptic patients.Results The prevalence of IDs in adult patients with epilepsy was 38.2% ,49.0% in active epilepsy and 12.1 %in seizure freedom. 30.0% ,5.5% ,and 2.7% were experiencing mild-to-moderate (HAMD score≥8),moderateto-severe ( ≥ 18 ) and severe ( ≥25 ) depression. 42 patients who met the HAMD score≥8 were classified as IDs group,and the remaining 68 patients were classified as non-IDs group. With multiple stepwise backward logistic regreasion, independent predictors of IDs were epileptic seizures ( OR = 8. 845, P = 0. 003 ); symptomatic or cryprogenic epilepsy ( OR = 3.132, P = 0. 045 ); prolonged duration of illness ( OR = 1. 106, P = 0.004 ) and employment status (OR =0. 154, P=0.001 ). There were no relationship between seizure frequency and severity of IDs ( Kruskal-Wallis test, x2 = 4.5, P = 0. 104). Conclusion IDs is a frequent psychiatric comorbidity in adult patients with epilepsy. The prevalence of IDs is higher in those with active epilepsy compared with those in seizure freedom and most of them are mild-to-moderate. Epileptic seizure, symptomatic or cryprogenic epilepsy, prolonged duration of illness and employment status are independent predictors of IDs, but seizure frequency has nothing to do with the IDs severity of patients.
9.Neuropsychological characteristics in active epilepsy and its risk factors
Yong HU ; Qing DI ; Nian YU ; Yanfang ZHANG ; Lingying SU
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(6):516-518
Objective To assess the neuropsychological characteristics in active epileptic patients and investigate itsrisk factors. Methods Ninety adult epileptic patients included 60 active epileptic patients (two or more unprovoked seizures within 12 months) and 30 age-, sex-, education-, course of disease- and seizure type-matched seizure-free subjects (without epileptic seizure for at least 1 year) . The neuropsychological tests including trail making test,digit symbol test, verbal fluency test,digit span test and hamilton depression scale( HAMD) ,were used to detect mental and motor speed, attention, language, working memory and depression symptoms respectively. The neuropsychological tests were compared between active and seizure-free epileptic patients and identified the risk factors of neuropsychological deficits in active epileptic patients. Results Compared to seizure-free subjects, active epileptic patients had significantly worse scores in digit symbol test, verbal fluency test, digit span test ((47.45 ±18. 812) vs(56.40 ±13. 631), (25. 25 ±8. 163) vs(30.40 ±8. 414), (10. 39 ±2. 228) vs( 11. 80 ± 2.074) respectively) ; more time to accomplish the trail making test A and B((64. 35 ±31.710) vs( 45. 47 ± 16. 309) , ( 133. 18 ± 47. 331 ) vs ( 98. 00 ± 35. 003 ) respectively) ; and higher scores in depressive symptoms ((9.12 ±6.219)vs(3.77 ±3.997) ,all P<0.05). Within active epileptic group,significant predictors of neuropsychological deficits were identified in a stepwise linear regression analysis: advancing age was significantly negatively correlated with digit symbol test(β = -0. 468, P = 0. 000) , digit span test (β = -0. 439, P = 0. 000), trail making test A (β =0.365, P = 0.003) and B(β = 0.346, P=0.002) ; higher scores on depressive symptoms was significantly negatively correlated with digit symbol test (β = -0.244, P = 0.015) ; mental work,high-education level and monotherapy were positively correlated with some of the cognitive function subscales. Conclusion This study suggests that active epilepsy can have a direct adverse effect on cognition and depression symptoms. Multi-drug therapy, severity of depression symptoms, advancing age, low-education level and non-mental work are the predictors of neuropsychological impairment in active epilepsy. In addition, good seizure control even after 1 year can have a beneficial impact on cognitive and depression prognosis.
10.Endoscopic implantation of 5-FU slow-release particles for advanced gastroenteric tumor
Nian FU ; Qing WU ; Xuefeng YANG ; Zhaoxia LIU ; Fei PENG
Clinical Medicine of China 2011;27(1):75-77
Objective To investigate the feasibility, safety and short-term efficacy of endoscopic implantation of 5-FU Slow-release Particles for advanced gastroenteric tumor. Methods During the endoscopy procedure,slow-releasing 5-FU agents were implanted densely into the tumors and infiltrated area. Forty-five to sixty pieces of agents ( each piece equivalent to 1.67 mg 5-FU)were injected, which containing an average dose of 100 mg 5-FU. Results A total of 13 advanced gastric cancer patients were enrolled into this study. Significant effects were observed in 3 patients and good effects in 8 patients, but 2 cases with no effects. The total effective rate was 84.62%. The endoscopy examination showed that the size of tumors reduced in various degree at 1 - 2months after the implantation. No hemorrhage or perforation was observed. Parameters of hepatorenal function and routine blood test were stable after implantation. Conclusion Endoscopic implantation of 5-FU Slow-release Particles can relieve the symptoms of patients and limit tumor growth in advanced gastroenteric tumor with no marrow and hepatorenal functional impair.