Expression and clinical significance of α1-antitrypsin in thyroid-associated ophthalmopathy / 上海交通大学学报（医学版）

Objective : To investigate the expression and clinical significance of α1-antitrypsin (α1-AT) in tears of thyroid-associated ophthal-mopathy (TAO) patients. Methods ¡¤ Patients diagnosed with TAO at Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January to December 2019 were included and divided into active TAO group (n=33) and inactive TAO group (n=22). Thirty eyes of 30 healthy volunteers were included (n=30). Unstimulated tear samples were collected from all subjects and the concentration of α1-AT was analyzed by enzyme-linked immunosorbent assay (ELISA). Difference of α1-AT concentration among 3 groups was compared by using Kruskal-Wallis H test. Mann-Whitney U test was performed for further comparison between each 2 groups. Correlationship between α1-AT con-centration and clinical activity score (CAS) was analyzed by Spearman relation. Statistical significance was accepted at a value of P<0.05. Re-sults ¡¤ α1-AT was significantly higher in active TAO group than that in inactive TAO group and control group (both P<0.05). There was no significant difference in α1-AT between inactive TAO group and control group. Tear α1-AT level was significantly correlated with CAS (r=0.846, P=0.000). ROC curve showed that the optimal diagnostic cut off value of α1-AT was 939.48 ng/mL, and the sensitivity and specificity were 81.8% and 100% (AUC=0.959, P<0.05). Conclusion ¡¤ α1-AT is significantly elevated in the tears of active TAO patients. Tear α1-AT level has important value in the diagnosis of clinical activity of TAO.

Effect of Tangnaikang on Imbalance Between NE and α1-AT in Spontaneous Type 2 Diabetic Mouse / 中国实验方剂学杂志

Objective: To observe the regulatory effect of Tangnaikang (TNK) on imbalance between neutrophil elastase (NE) and α1-antitrypsin (α1-AT) in ob/ob mice with type 2 diabetes mellitus (T2DM). Method: Thirty-two male SPF ob/ob mice were randomly divided into model group (DM, normal saline) and high-dose TNK group (TNKH, TNK solution 16.04 g·kg-1), middle-dose TNK group (TNKM, TNK solution 8.02 g·kg-1) and low-dose TNK group (TNKL, TNK solution 4.01 g·kg-1). Another 8 C57BL/6J mice were included in normal group (Con, saline). The experiment lasted for four weeks. The general state, body weight (BW) and fasting blood glucose (FBG) of the mice were recorded weekly, the oral glucose tolerance (OGTT) test was performed on the 25th day, the insulin tolerance (ITT) test was performed on the 27th day, and the area under the curve (AUC) was calculated. After the end of the experiment, serum was used to detect the level of fasting insulin (Fins), insulin resistance index (HOMA-IR), total triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), NE and α1-AT. Adipose tissue was used to detect the expressions of NE, α1-AT, phosphor-insulin receptor substrate 1 antibody (p-IRS1) and glucose transporter 4 (GLUT4) proteins. Result: Compared with the Con group, the BW of the ob/ob mice of the model group increased significantly, the glucose and lipid metabolism indexes showed diabetes, the serum and adipose tissue NE increased significantly (Pα1-AT decreased significantly (PPPogtt and AUCITT were significantly decreased (PPα1-AT increased significantly (PPConclusion: TNK can reduce the BW of ob/ob mice, improve glycolipid metabolism, increase α1-AT level, decrease NE level, and regulate IRS1-GLUT4 signaling pathway, which may be one of its mechanisms in improving IR of adipose tissue mediated by neutrophil.

Research progress ofα1-antitrypsin deficiency of mutant Z / 上海交通大学学报（医学版）

α1-Antitrypsin (α1-AT) belongs to serine protease inhibitor (Serpin) superfamily and is the main protease inhibitor in human circulation. It can inhibit many proteases to protect tissues from digradation. The mutant Z (Glu342Lys) of α1-AT predisposes to the early onset of emphysema due to decreased functional α1-AT in the lung and to neonatal hepatitis due to accumulation of α1-AT polymers in the endoplasmic reticulum of hepatocytes, which disrupts the balance between protease and protease inhibitors. This paper reviews recent research progress on the pathogenic mechanism and the prognosis of α1-antitrypsin deficiency.

Effect of single-hole thoracoscopic surgical treatment on elderly spontaneous ;pneumothorax patients’ postoperative assessment and serological markers / 中国医学装备

Objective: To analyze the effect of single-hole thoracoscopic surgical treatment on elderly spontaneous pneumothorax patients’ postoperative assessment and serological markers and explore single-hole thoracoscopic surgery application. Methods:78 cases of elderly patients with spontaneous pneumothorax were retrospectively analyzed, and divided into single-hole thoracoscopy group with 35 patients and conventional thoracoscopic group with 43 patients. Then the surgery indicators, postoperative pain scores and perioperative serum target level differences were compared between the two groups. Results: The operative time, postoperative drainage time and total hospital stays of single-hole thoracoscopic surgery patients were shorter than that of conventional thoracoscopic group. The differences were statistically significant (t=7.183, t=5.294, t=6.938;P<0.05). The blood loss and postoperative drainage were less. The differences were statistically significant (t=6.883, t=8.735; P<0.05). The postoperative pain scores within one week was less than conventional thoracoscopic group. The differences were also statistically significant (t=6.882, t=5.724, t=5.284;P<0.05). The serological indicators, CRP, IL-1, TNF-α,α1-AT levels of patients in the single-hole thoracoscopy group were lower than the conventional group (t=8.293, t=7.274, t=9.284, t=7.183;P<0.05). Conclusion:VATS and conventional hole thoracoscopic surgery can be used for treatment of spontaneous pneumothorax in elderly, but single-hole thoracoscopy has more advantages in reducing surgical trauma and postoperative pain and optimize systemic inflammation associated factors.

Efficacy and observation of Bufei Huoxue soup combined with Western medicine for stable COPD and airway inflammation reduction and lung function improve-ment / 中国免疫学杂志

Objective:To analyze Bufei Huo Xue Tang combined with Western medicine for chronic obstructive pulmonary disease( COPD) and its effect on airway inflammation reduction and lung function improvement. Methods: 100 patients with stable COPD in our department from February 2013 to March 2015 were randomly divided into combination group and control group. The control group received conventional oxygen,salmeterol and fluticasone propionate inhalation powder(1 absorption /times,2 times /day) and expectorant treatment. Combination group were based on the addition of lungs Bufei Huo Xue Tang with a course of 12 weeks. The clinical efficacy and α1-AT and IL-8 levels in two groups of patients were compared. Results:The cough,sputum,difficulty breathing, wheezing,wheezing symptom score in the combined group after treatment was significantly lower than the control group ( P<0. 05 ) . FVC,FEV1 and FEV1 /FVC in the combined group after treatment was significantly lower than those in the control group(P<0. 05).α1-AT and NO in the combination group was significantly higher after treatment. IL-8 and TNF-α was significantly lower than the control group(P<0. 05). BODE scores in the combined group after treatment were significantly lower than the control group(P<0. 05). Conclusion:Bufei Huoxue soup combined with Western medicine for COPD can inhibit inflammation,improve clinical symptoms,lung function and quality of life.

Detection ofα1 antitrypsin activity by chromogenic substrate assay with initial veloci-ty of enzymatic reaction / 军事医学

Objective To detect the activity of α1 antitrypsin(AAT) with initial velocity of enzymatic reaction in order to detect the activity of samples in the process of separating and purifying plasma protein ,chromogenic substrate assay was optimized.Methods The effect of trypsin concentration and reaction time on enzymatic reaction was acquired by the kinetic monitoring mode of the microplate reader .Initial velocity was calculated to confirm the largest concentration of trypsin which was saturated by substrate .AAT was incubated with trypsin and absorbance produced by enzymatic reaction of remaining trypsin and substrate could reflect the activity of AAT .A standard curve was established with △D fitting with the activity of AAT standard.The activity of related samples was detected and the precision and accuracy of the method were evaluated . Results Trypsin concentration was 0.0625 mg/ml.Within 20 minutes, enzymatic reaction was with initial velocity .The range of the standard curve was 200-1200 IU/ml.Correlation coefficient was more than 0.99.The activity of Cohn Ⅳ, samples of pre-processing and elution were (720.59 ±18.63), (601.84 ±19.18),and (568.09 ±24.83)IU/ml, respec-tively.The relative standard deviation was less than 10%. Sample recovery rate was 90%-110%.Conclusion The optimized chromogenic substrate assay greatly improves accuracy and precision .The method can be used for the detec-tion of AAT activity of samples in laboratories and workshops .

Effect ofα1-antitrypsin Z variant overexpression on autophagy regulation / 中国药理学通报

Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.

Acquired alpha 1-antitrypsin deficiency in tropical pulmonary eosinophilia.

Background & objectives: Observation of an increased frequency of an intermediate deficiency of serum alpha1-antitrypsin (α1-AT) in patients with Tropical Pulmonary Eosinophilia (TPE) was earlier reported. Though the possibility of existence of an acquired deficiency was suggested, without phenotyping a hereditary α1-AT deficiency in TPE could not totally be ruled out. In this study, we have done Pi (Protease inhibitor) phenotyping to investigate the possibility of association of any heterozygous (or homozygous) α1-AT deficiency in patients with TPE. Methods: Serum a1antitrypsin (α1-AT) was measured in 103 patients (Group A) with TPE, 99 patients with pulmonary eosinophilia who had associated intestinal worm infestation (Group B) and 43 healthy volunteers who served as controls. In 19 α1-AT deficient patients (9 of Group A and 10 of Group B), α1-AT level was measured before and after treatment. In 58 patients with TPE and in 5 controls, phenotyping was done. Results: Fifteen patients of Group A and 16 from Group B showed intermediate α1-AT deficiency (150 mg % or less. None of the control subjects had α1-AT deficiency (<200 mg%). After treatment with DEC and/or deworming, in 19 patients there was a significant (P < 0.001) rise in α1-AT levels. Results of phenotyping showed that all had M1 or M2 allele and none had S or Z variant (either homozygous or heterozygous) thus ruling out any underlying genetic cause for the observed α1-AT deficiency. Interpretation & conclusions: The observed α1-AT deficiency may be due to the chronic inflammation in TPE and associated oxidative stress. However, in such α1-AT deficient patients with TPE and those with worm infested pulmonary eosinophilia, faecal α1-AT concentration and faecal α1-AT clearance should be routinely estimated to rule out the possibility of any intestinal protein loss.

Investigated the serum α1-antitrypsin concentrations in 1352 petrochemical workers / 国际检验医学杂志

Objective To observe the effect of atmospheric pollution on workers'α1-antitrypsin concentrationb in petrochemical enterprises.Methods During health examination,the workers were indivded into 3 groups according tO the length of job experience.Ⅰ group(<10 years)included 291 cases,aged 22-35 years old,Ⅱ group(10-20 years)included 634 cases,aged 33-45 years old,and Ⅲgroup(>20 years)included 427 cases,aged 44-55 years old.The normal control were individed int different group according to the status of petrochemical workers.,all the serum samples were detected on automatic biochemical analyzer,α1-antitrypsin levels were analyzed between the different groups.Results In group Ⅰ,the α1-AT concentration was 2.58±0.38 g/L(P>0.05)without significant difference compared with control.In group Ⅱ and Ⅲ,the concentration α1-AT were 2.02±0.43 g/L and 1.72±0.41 g/L respectively with significant differences compared with control.And the significant difference existed among the three groups of petrochemical workers..But the sigficant difference did not exist between different group of control people.Conclusion During the process of oil refining,the plants exhausted the polluted gas of nitrogen dioxide,olefins,alkanes,alcohols,carbonyl and other harmful substances,it influenced theα-AT levels of body.It is a good suggestion to decreas the petrochemical pollution,and to monitor the serum concentration of α-AT content among petrochemical workers,it might to reduce the incidence of lung disease of great significance.

Protective effects of α-1 antitrypsin on human islets injury after transplantation by pancreas exocrine cells / 中华器官移植杂志

Objective To investigate the protective effects of α-1 antitrypsin on human islets injured by protease released from pancreas exocrine cells. Methods ( 1 ) in vivo experiment. Parts of the cadaveric pancreas was digested with collagenase, islets were selected artificially, and pancreatic exocrine cells were collected. 8-9 weeks olds male BALB/c-Nu nude mice were induced into diabetic mice with STZ (240 mg/kg body weight, i. p) and randomly divided into two groups: the control group (n = 6), 250 islets were transplanted into left kidney subcapsule of diabetic nude mice; cotransplant group (n = 7), 250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule. Blood glucose level was monitored. Nephrectomies were performed after 28 days. The expression of anti-amylase antibodies in subcapsule was detected by using immunohistochemical staining. (2) Islets culture: Three groups were randomly set up. Group 1: purified islet group, 250 islets were incubated into a 6-well culture plate; Group 2: non-purified islet group, 250 purified islets and equal volume of exocrine cells were incubated; Group 3: nonpurified islet + Al AT group, 250 purified islets and equal volume of exocrine cells were incubated with α-1 antitrypsin added (0. 5 mg/ml). After 48 h, insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured. Results 10000 islets were collected.After islets transplantation, the blood glucose levels in control and co-transplant groups were normal,but a delayed islet function in reversing diabetes was in the co-transplant group, and ehe mice in both groups became hyperglycemic after nephrectomy. A large number of anti-amylase antibody-positive cells were found in renal subcapsule in the co-transplant group while little seen in the control group.Insulin levels in the non-purified islet group were decreased as compared with purified islet group,those in the non-purified islet group + A1AT group were higher than in the non-purified islet group,but lower than in the purified islet groups. Trypsin concentration in the non-purified islet group was increased as compared with purified group, that in the non-purified islet group + A1AT group was lower than the non-purified islet group, but higher than in the purified islets group (all P＜0. 01).Conclusion Protease released from acinar cells during pancreatic digestion has detrimental effect on islet function after transplantation. Co-cultivation of islets and pancreatic exocrine cells with A1AT added can prevent islet cell damage caused by trypsin.

Prevalence of alpha1-Antitrypsin Genotypes in Koreans / 결핵

BACKGROUND: Alpha-1-antitrypsin (A1AT) deficiency is the only established genetic resk factor for emphysema. This study was undertaken to investigate the prevalence of the genotypes of A1AT genotypes in healthy Koreans. METHOD: The study population consisted of 380 healthy Koreans enrolled at the Health Promotion Center in Kyungpook National University Hospital. The polymerase chain reaction (PCR) and restriction fragment length polymorphim (RFLP) for detecting the A1AT variants M1(Ala), M1(Val), M2, S and Z were used. RESULTS: The genotypes of subjects were as follows : M1(Val)/M1(Val), 254(66.8%) ; M1(Val)/M2, 105(27.6%) ; M2/M2, 19 (5.0%) ; and M1(Val)/M1(Ala), 2 (0.5%). There was no case with 'deficiency' alleles such as S and Z found in this study. CONCLUSION: These results suggest that A1AT deficient alleles are either extremely rare or not present in Koreans.