1.Purification of L-amino acid oxidase from Naja atra venom and its effects on endothelial cells
Zhou CHEN ; Jianjun HUANG ; Ling XUE ; Yunlu XU ; Lishan LIN
Chinese Pharmacological Bulletin 2003;0(08):-
Aim To purify L-amino acid oxidase(LAAO) from the venom of Naja atra and study its effect on endothelial cells.Methods The NAV-LAAO was purified by ion-exchange chromatography and affinity chromatography.The MTT assay and Western blot were used to detect the viability and apoptosis of HUVEC.The tubule-forming was used to study the angiogenesis of cells.Results The NAV-LAAO was purified successfully from the venom of Naja atra.The molecular weight of NAV-LAAO was determined to be 58 ku by SDS-PAGE.NAV-LAAO effectively inhibited the growth and tubule-forming of HUVEC,and the 50% inhibitory concentration(IC50) was 21.42 mg?L-1.Compared with control,the levels of caspase-3 and caspase-8 increased in HUVEC treated with NAV-LAAO.Conclusions The NAV-LAAO is purified successfully from Naja atra venom by ion-exchange chromatography and affinity chromatography.The NAV-LAAO inhibits the growth and tubule-forming capacity of HUVEC in a dose-dependent manner.
2.Clinical observation of therapeutic effect of external application of Chinese herbal medicine film plus acupoint-injection on breast hyperplasia
Youyi CHEN ; Lishan HUANG ; Lisheng CHEN ; Ping WANG ; Ping LIN
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(12):-
Objective:To observe the therapeutic effect ofexternal application ofChinese herbal medicine film plus acupoint-injection on breast hyperplasia.Methods:The 120 cases ofbreast hyperplasia with syndrome ofthe liver failing to maintain the normal flow ofqi were randomly divided into treatment group(40cases) which was treated by external application ofChinese herbal medicine film plus acupoint-injection, acupoint-injection group(40 cases) and control group(40cases).The changes ofsymptoms and signs, level ofserum sexual hormone and color doppler ultrasound analysis ofthree group were observed for comparison.Results:After 1 course oftreatment, the total effective rate oftreatment group was obviously higher than that ofcontrol group(P0.05).After 2 courses oftreatment, the total effective rate oftreatment group and acupoint-injection group was obviously higher than that ofcontrol group(P
3.Control study on skull to boneless head CTA by surface shaded display technique and volume computed tomographic digital subtraction angiography in diagnosing AVM
Wei LIN ; Huamin TANG ; Boli ZHANG ; Lishan WANG ; Bao CHEN
Chongqing Medicine 2016;45(11):1508-1510,1514
Objective To investigate the diagnostic value of technologies for arteriovenous malformation(AVM ) ,which are skull to boneless head CTA by surface shaded display technique (hereinafter referred to as the SSD‐CTA technology) .Methods Totally 27 patients of the First Affiliated Hospital of Chongqing Medical University were selected ,which were diagnosed with AVM by VCTDSA .The original data to our hospital CT workstation were imported ,SSD‐CTA technology to bone reconstruction were performed ,and the data of the two groups were compared .Another 5 patients in our hospital which were diagnosed with AVM by SSD‐CTA examination were selected ,and compared the results with DSA .Results The quality score of SSD‐CTA reconstructed image was less than that of VCTDSA ,but the diagnosis of the two technologies for AVM had no significant statistical difference (P>0 .05) .The number of AVM patients from our hospital wsa too less to do the statistical analysis .The detail resolution of SSD‐CTA was less than DSA ,but the diagnosis of SSD‐CTA was consistent with DSA .Conclusion SSD‐CTA has clinical practical val‐ue ,can be used as a check ,intracranial venous malformation diagnosis .
4.Clinical significance of urinary mALB ,RBP and CysC and their combined detection in early diagnosis of type 2 diabetic nephropathy*
Shaoxiong ZHOU ; Yongjian SHEN ; Lin ZOU ; Lishan HUANG
International Journal of Laboratory Medicine 2017;38(6):786-787,791
Objective To investigate the application value of urine microalbumin (mALB) ,retinol binding protein(RBP) and cys-tatin C(CysC) and their combined detection in early diagnosis of type 2 diabetic nephropathy(DN) .Methods Ninety-two inpatients with DN (DN group) and 90 people undergoing the physical examination(control group) in our hospital from June 2014 to Decem-ber 2015 were collected .Urine mALB ,RBP and CysC were detected in all subjects and detection results were analyzed statistically . Results The levels of urine mALB ,RBP and CysC in the DN group were significantly higher than those in the control group ,the differences all had statistical significance (P< 0 .05) .Among 3 indicators ,the positive rate of urine mALB for detecting DN was highest (94 .57% ) ,while which of 3-index combined detection was 97 .83% ,and significantly higher than that of single detection , the difference was statistically significant(P<0 .05) .The sensitivity ,specificity ,positive predictive value ,negative predictive value and Youden index of 3-index combined detection were all higher than those of single index .The ROC curve showed that AUC of u-rine mALB for diagnosing DN was 0 .732 ,the diagnostic cut-off value was 43 .58 mg/L ,AUC of urine RBP was 0 .685 ,the diagnos-tic cut-off value was 1 .47 mg/mL ,AUC of urine CysC was 0 .701 ,the diagnostic cut-off value was 1 .42 mg/L ,while AUC of com-bined detection was 0 .928 .Conclusion Urine mALB ,RBP and CysC are better indexes reflecting renal injury .Their combined de-tection will increase the positive rate ,sensitivity and specificity for diagnosing DN .So monitoring the levels of urine mALB ,RBP and CysC has an important significance to diagnosing the occurrence and development of DN early renal injury and prevention ,treat-ment and delaying progress of DN .
5.Simultaneous determination of amlodipine, benazepril and benazeprilat in human plasma by LC-HESI/MS/MS method.
Hualing PAN ; Lishan LIN ; Juefang DING ; Xiaoyan CHEN ; Dafang ZHONG
Acta Pharmaceutica Sinica 2014;49(1):95-100
The study aims to develop a rapid, sensitive and specified method of liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) for simultaneous determination of amlodipine, benazepril and benazeprilat in human plasma using amlodipine-d4 and ubenimex as internal standards (ISs). Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the positive mode for mass spectrometric detection. Analytes and ISs were extracted from plasma by simple protein precipitation. The reconstituted samples were chromatographed on a C18 (100 mm x 4.6 mm, 5 microm) column with mixture of methanol-acetonitrile-5 mmol.L- ammonium acetate-formic acid (30 : 30 : 40 : 0.1) as mobile phase at a flow rate of 0.6 mL.min-1. The standard curves were demonstrated to be linear in the range of 0.02 to 6.00 ng.mL-1 for amlodipine, 0.2 to 1,500 ng.mL-1 for benazepril and benazeprilat with r2>0.99 for each analyte. The lower limit of quantitation was identifiable and reproducible at 0.02, 0.2 and 0.2 ng mL-1 for amlodipine, benazepril and benazeprilat, respectively. The intra-day and inter-day precision and accuracy results were within the acceptable limit across all concentrations. The plasma samples were stable after four freeze-thaw cycles and being stored for 93 days at -20 degrees C. The method was applied to a pharmacokinetic study of a fixed-dose combination of amlodipine and benazepril on Chinese healthy volunteers.
6.Comparing the estimated glomerular filtration rate through different equations in evaluating kidney function of the population
Lishan SUN ; Liu LU ; Mingdong WANG ; Lin LU ; Ming YU ; Lieying FAN
International Journal of Laboratory Medicine 2015;(10):1370-1372,1375
Objective To compare the difference of kidney function evaluated by using 3 different estimated glomerular filtration rate(GFR) equations in populations .Methods Retrospectively analyzed 65 856 patients who measured serum creatinine and Cysta‐tin C at the same time ,and come from the outpatients or inpatients of the hospital .The estimated GFR (eGFR) were calculated through 3 equations ,then compared the eGFR in the population and among different groups according to different kidney functions , and then grouped the people enrolled in the study again according to the eGFR calculated by using the 3 different equations and compared the differences among groups .Results Compared with the eGFR calculated by using Creatinine equation ,the correlation coefficients of the eGFRs calculated by using the other two equations were 0 .81 and 0 .90 ,respectively ,both P<0 .05 ;The differ‐ence between the means of eGFR were 6 .19 and 1 .79 mL/(min × 1 .73 m2 ) respectively with obvious significance (P<0 .01) ,in consistency analysis .There were obvious overestimation of kidney function when using Creatinine equation to calculate eGFR .Con‐clusion There is consistence and obvious difference by using the 3 CKD‐EPI′s eGFR equations .Physicians should choose suitable equations to evaluate kidney function in different populations .
8.Clinical significance of detecting glucagon like peptide-1 in late-onset Alzheimer′s disease
Mingdong WANG ; Lishan SUN ; Ming ZONG ; Liu LU ; Lin LU ; Lieying FAN
Chinese Journal of Laboratory Medicine 2015;(8):543-547
Objective To explore the serum level of Glucagon like peptide-1 in late-onset Alzheimer′s patients and its clinical significance.Methods Case control study.Collecting cerebral vascular disease fifty-five cases, diagnosed with late-onset Alzheimer′s disease sixty-one cases, type 2 diabetes mellitus fifty-one cases , type 2 diabetic patients combined with late-onset Alzheimer′s disease thirty-seven patients from the Shanghai East Hospital and partly Pudong area elderdly hospital during October 2013 to March 2014, and forty healthy persons as normal control from physical examination center of Shanghai East Hospital during September 2013 to February 2014.Measuring the concentrations of GLP-1,β-amyloid, Tau protein and other routinely used clinical tests in the serum of patients from the normal controls , cerebrovascular disease , late-onset Alzheimer′s disease, type 2 diabetes and type 2 diabetes mellitus combined with late-onset Alzheimer′s disease by ELISA method developed in our laboratory.The blood samples were also collected at three fixed time including fasting time ,1 hour after taking glucose , 2 hour after taking glucose, the concentrations of GLP-1 were determined in the LOAD group , T2DM group and the T2DM combined with LOAD group and normal control group.The concentrations of serum GLP-1 among groups were compared with single factor analysis of variance , and the concentrations of serum GLP-1 between the two groups were compared using LSD-t test.Analysing the correlation between GLP-1 and other indicators with Pearson analysis.Results The fasting GLP-1 levels of LOAD group were ( 123.4 ±20.8 ) nmol/L, and they were highest between the normal control group (78.6 ±6.0) nmol/L and the cerebral blood vessel disease group(89.0 ±8.7)nmol/L (F values were 3.46 and 1.98, P<0.05).The fasting GLP-1 levels of T2DM combined with LOAD group (157.9 ±28.6) nmol/L were higher than the LOAD group (123.4 ± 20.8) nmol/L (t =1.63,P <0.05), but there were no difference of the fasting GLP-1 levels between T2DM combined with LOAD group (157.9 ±28.6) nmol/L and T2DM group(153.8 ±18.0)nmol/L(t=0.96,P>0.05).Deficient secretion of GLP-1 after taking glucose 1 hour in most of the patients of T2DM combined with LOAD group (99.1 ±14.2) nmol/L, LOAD group(73.9 ±6.6 ) nmol/L and T2DM group (96.3 ±7.0 ) nmol/L could be concluded .The GLP-1 levels of T2DM combined with LOAD group after taking sugar 2 hour were (115.4 ±18.6)nmol/L ,and were higher than that of normal levels (63.3 ±6.2) nmol/L after taking sugar 2 hour(t=4.49,P<0.05).There were no difference between the GLP-1 levels of the LOAD group (73.6 ±5.8 )nmol/L and the GLP-1 levels of the normal group(63.3 ±6.2)nmol/L after taking sugar 2 hour (t=0.94,P>0.05).Pearson correlation analysis showed that the relationship of the levels of GLP-1 with Aβ( 1-42 ) and the levels of GLP-1 after taking glucose 1 h and 2 h were positively relative, and its coefficients of correlation were 0.401,0.436,0.722.Conclusions LOAD and T2MD are similar, and they have GLP-1 secretion shortage phenomenon after taking glucose , so monitoring dynamic change of GLP-1 after taking glucose may contribute to the auxiliary diagnosis of LOAD.
9. The relationship between bolus volume and hyoid displacement in dysphagia patients with nasopharyngeal carcinoma after radiation therapy
Lishan CHEN ; Huichang ZHOU ; Pande ZHANG ; Chuke LIN ; Peng LIANG ; Zhiyong GUAN ; Jiajian YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2019;41(12):894-899
Objective:
To evaluate the relationship between bolus volume and hyoid displacement in dysphagia patients with nasopharyngeal carcinoma after radiation therapy.
Methods:
Twenty-three nasopharyngeal carcinoma patients with dysphagia were recruited and their swallowing of 3, 5, 10 and 20ml of liquid food was studied fluoroscopically. The vertical and horizontal displacement of the hyoid as well as its time in motion were measured, and the relationship between the bolus volume, hyoid displacement and time in motion time was evaluated.
Results:
The largest vertical displacement of the hyoid (1.01±0.65cm) was observed when swallowing a 10ml bolus. The hyoid showed the smallest average horizontal displacement (0.39±0.34cm), when swallowing a 3ml bolus. The average motion time of the hyoid was (2.11±0.65) seconds. It was shorter when swallowing a 10 or 20ml bolus than when dealing with a smaller one. Hyoid motion time was negatively correlated with the horizontal displacement of the hyoid bone, and the volume of a swallow was negatively correlated with the hyoid motion time but positively correlated with the penetration-aspiration scale score.
Conclusion
Bolus volume affects hyoid displacement and hyoid motion time in nasopharyngeal carcinoma patients with dysphagia after radiation therapy. For patients with a penetration-aspiration scale score of 5 or less, the optimum bolus volume is 5 to 10ml.
10.Advance of mitophagy in diabetic cardiomyopathy
Cuihua HUANG ; Lu LIN ; Lishan HUANG ; Libin LIU
Chinese Journal of Endocrinology and Metabolism 2023;39(6):522-526
Diabetic cardiomyopathy is a myocardial complication associated with abnormal glucose metabolism and dyslipidiaemia, which increases the risk of death and heart failure in diabetic patients. Mitochondrial dysfunction is involved in the occurrence and development of diabetic cardiomyopathy. Recent studies have confirmed that scavenging damaged mitochondria in cardiomyocytes through mitophagy can restore mitochondrial homeostasis, reduce oxidative stress and improve diabetic cardiomyopathy. Therefore, this article provides a comprehensive review of the mechanisms and characteristics of mitochondrial autophagy in diabetic cardiomyopathy. It aims to offer new insights and theoretical basis for the prevention and treatment of diabetic cardiomyopathy.