Alpha-1 antitrypsin deficiency and Pi^(*)Z allele as important co-factors in the development of liver fibrosis

BACKGROUND Alpha-1 antitrypsin deficiency(AATD)is a codominant autosomal hereditary condition that predisposes patients to the development of lung and/or liver disease,and Pi*Z allele is the most clinically relevant mutation.AIM To evaluate the impact of clinical parameters and AATD phenotypes,particularly the Pi*Z allele,in liver fibrosis.METHODS Cross-sectional cohort study including consecutive patients with AATD followed in Pulmonology or Hepatology consultation.RESULTS Included 69 patients,49.3%had Pi*MZ phenotype and 10.1%Pi*ZZ.An age≥55 years,age at diagnosis≥41 years and AAT at diagnosis<77 mg/dL predicted a nonalcoholic fatty liver disease fibrosis score(NFS)not excluding advanced fibrosis[area under the curve(AUC)=0.840,P<0.001;AUC=0.836,P<0.001;AUC=0.681,P=0.025].An age≥50 years and age at diagnosis≥41 years predicted a fibrosis-4 index of moderate to advanced fibrosis(AUC=0.831,P<0.001;AUC=0.795,P<0.001).Patients with hypertension,type 2 diabetes mellitus(DM),dyslipidaemia,metabolic syndrome,and regular alcohol consumption were more likely to have a NFS not excluding advanced fibrosis(P<0.001,P=0.002,P=0.008,P<0.001,P=0.033).Patients with at least one Pi*Z allele and type 2 DM were 8 times more likely to have liver stiffness measurement≥7.1 kPa(P=0.040).CONCLUSION Risk factors for liver disease in AATD included an age≥50 years,age at diagnosis≥41 years,metabolic risk factors,regular alcohol consumption,at least one Pi*Z allele,and AAT value at diagnosis<77 mg/dL.We created an algorithm for liver disease screening in AATD patients to use in primary care,selecting those to be referred to Hepatology consultation.

Collagen-Ⅵ-alpha-1在胶质母细胞瘤中的表达及临床意义

目的探讨Collagen-Ⅵ-alpha-1(COL6A1)在胶质母细胞瘤中的表达以及与临床病理特征的关系。方法采用免疫组织化学方法检测35例胶质母细胞瘤组织和22例正常脑组织中COL6A1的表达,并对其表达进行临床病理特征分析。结果COL6A1在胶质母细胞瘤中阳性表达率为88.57%(31/35),而在正常脑组织中仅为9.09%(2/22),差异具有统计学意义(P<0.05)。COL6A1的表达在不同性别、不同年龄、不同KPS评分的患者中差异无统计学意义(均P>0.05),但在不同体积的肿瘤及是否完整切除患者中的差异有统计学意义(P<0.05)。COL6A1阳性表达的胶质母细胞瘤患者总生存期明显短于阴性表达的患者(P<0.05)。结论 COL6A1可能与胶质母细胞瘤的发生发展密切相关,有望成为胶质母细胞瘤诊断治疗的新靶点。

Efficacy of thymosin alpha-1 and interferon alpha in treatment of chronic viral hepatitis B:A randomized controlled study

AIM： To observe the efficiency and safety of thymosin-α1 treatment in patients with hepatitis B e antigen （HBeAg） and HBV DNA positive chronic hepatitis. METHODS： Sixty-two patients were randomly divided into groups A and B. The patients in group A received subcutaneous injection of 1.6 mg thymosin-α1, twice a week （T-α1 group） for six months, and the patients in group B received 5 MU interferon alpha （IFN-α） each day for fifteen days, then three times weekly （IFN-α group） for six months. The results between two groups treated with and the group untreated with IFN-α which was followed up for 12 mo （historical control group consisting of 30 patients） were compared, and three groups were comparable between each other （P 〉 0.05） at baseline （age, sex, clinical history, biochemical, and serological parameters）. RESULTS： At the end of treatment, complete response, which was defined as alanine aminotransferase （ALT） normalization and HBV DNA and HBeAg loss, occurred in 9 of 29 （31.0%） patients in the T-α1 group and in 15 of 33 （45.5%） patients in the IFN-α group （χ^2= 1.36, P 〉 0.05）. After a follow-up period of six months, a complete response was observed in 14 of 29 （48.3%） patients in the T-α1 group and in 9 of 33 （27.3%） patients in the IFN-α group （χ^2= 2.93, P 〉 0.05）. Compared with the results observed in the historical control （HC） group untreated with IFN-α which was followed up for 12 mo, the rate of complete response was significantly higher in IFN-α group at the end of therapy （1 of 30 vs 15 of 33, 7：2 = 14.72, P 〈 0.001） and in the T-α1 group at the end of follow-up （1 of 30 vs 14 of 29,χ^2 = 15.71, P 〈 0.001）. In T-α1 and IFN-α treatment groups, the area under （the plasma concentration time） curve （AUC） of negative HBV DNA and HBeAg was 340, 17%, 31% and 19% smaller than that in the HC group. By the end of the follow-up period, the proportions of ALT normalization and negative HBV DNA in the T-α1 group were significantly higher than those in the IFN-α and HC groups. The odds of ALT normalization and negative HBV DNA at the end of the follow-up was three-fold higher in the T-α1 group than in the IFN-α group. Unlike IFN-α, T-α1 was well tolerated by all patients, and no side effects appeared in T-α1 group. CONCLUSION： The results suggest that a 6-too course of T-α1 therapy is effective and safe in patients with chronic hepatitis B. T-α1 is able to reduce HBV replication in patients with chronic hepatitis B. Furthermore, T-α1 is better tolerated than IFN-α and can gradually induce more sustained ALT normalization and HBV DNA and HBeAg loss. However, a response rate of 48.3% is still less ideal. A more effective therapeutic approach warrants further study.

Alpha-1 antitrypsin deficiency and the risk of hepatocellular carcinoma in end-stage liver disease

AIM:To evaluate the association between alpha-1 antitrypsin deficiency(A1ATD) and hepatocellular carcinoma(HCC) in patients with end-stage liver disease(ESLD).METHODS:Patients with cirrhosis and ESLD referred to the Cleveland Clinic Foundation for liver transplantation between 2003 and 2014 were included in the study(N = 675). ESLD was defined as having histological features of cirrhosis and/or radiological evidence of cirrhosis in the context of portal hypertension(ascites,variceal bleeding,thrombocytopenia,or hepatic encephalopathy). A1 ATD was diagnosed using phenotype characterization(MZ or ZZ),liver biopsy detection of PAS-positive diastaseresistant(PAS+) globules,or both. Patients with other causes of liver diseases such as hepatitis C virus(HCV),alcoholic liver disease and non-alcoholic steatohepatitis(NASH) or NASH were also included in the study. HCC was diagnosed by using imaging modalities,biopsy findings,or explanted liver inspection. Follow-up time was defined as the number of years from the diagnosis of cirrhosis to the diagnosis of hepatocellular carcinoma,or from the diagnosis of cirrhosis to the last follow up visit. The rate of HCC was assessed using time-tointerval analysis for interval censored data.RESULTS:This study included 675 patients. 7% of subjects had A1ATD(n = 47). Out of all subjects who did not have A1 ATD,46% had HCV,17% had alcoholic liver disease,19% had NASH and 18% had another primary diagnosis. Of the 47 subjects with A1 ATD,15 had a primary diagnosis of A1ATD(PI*ZZ phenotype and PAS+ globules),8 had a PI*MZ phenotype alone,14 had PAS+ alone,and 10 had both the PI*MZ phenotype and PAS+. Median follow-up time was 3.4(25th,75 th percentiles:1,5.2) years. The overall rate of hepatocellular carcinoma in all subjects was 29%(n = 199). In the A1 ATD group,the incidence rate of HCC was 8.5% compared to 31% in the group of patients with other causes of cirrhosis(P = 0.001). Patients with ESLD due to A1 ATD had the lowest yearly cumulative rate of hepatocellular carcinoma at 0.88% per year compared to 2.7% for those with HCV cirrhosis,1.5% in patients with NASH and 0.9% in alcohol-induced liver disease(P < 0.001).CONCLUSION:Within this group of patients with ESLD,there was no significant association between A1 ATD and increased risk of HCC.

Delayed diagnosis of alpha-1-antitrypsin deficiency following post-hepatectomy liver failure: A case report

Post-hepatectomy liver failure(PHLF) is a leading cause of morbidity and mortality following major liver resection. The development of PHLF is dependent on the volume of the remaining liver tissue and hepatocyte function. Without effective pre-operative assessment, patients with undiagnosed liver disease could be at increased risk of PHLF. We report a case of a 60-year-old male patient with PHLF secondary to undiagnosed alpha-1-antitrypsin deficiency(AATD) following major liver resection. He initially presented with acute large bowel obstruction secondary to a colorectal adenocarcinoma, which had metastasized to the liver. There was no significant past medical history apart from mild chronic obstructive pulmonary disease. After colonic surgery and liver directed neo-adjuvant chemotherapy, he underwent a laparoscopic partially extended right hepatectomy and radio-frequency ablation. Post-operatively he developed PHLF. The cause of PHLF remained unknown, prompting reanalysis of the histology, which showed evidence of AATD. He subsequently developed progressive liver dysfunction, portal hypertension, and eventually an extensive parastomal bleed, which led to his death; this was ultimately due to a combination of AATD and chemotherapy. This case highlights that formal testing for AATD in all patients with a known history of chronic obstructive pulmonary disease, heavy smoking, or strong family history could help prevent the development of PHLF in patients undergoing major liver resection.

DETECTION OF ALPHA-1 ANTICHYMOTRYPSIN IN HEPATOCELLULAR CARCINOMA TISSUE

One hundred and fifty-three consecutive cases of HCC and 25 controls from autopsy material were studied by immunohistochemical method in this paper. A review of the histopathology and demonstration of AFP, alpha- 1-antichymotrypsin (AACT), alpha 1-antitrypsin (AAT) and CEA were made.Among the tumor markers. AACT yielded the highest positive rate, 109 cases (71%) out of 153 HCC. CEA was the next, 95 cases (62%) .AFP and AAT gave the same result, 72 cases (47%) . AACT, AAT and CEA were not found in the controls. AFP was present in a few hepatocytes in 1 of 25 controls. The results were in keeping with serum tests so far as the highest positive rate being AACT was concerned. Therefore, combined determination of AACT and AFP would seem a better screening method than by that of AFP alone for survey of HCC.

On the Activities of Pancreatic Proteases and Alpha-1 Proteinase Inhibitor in Meat-Type Chicken

The study was aimed at the evaluation of the effects of breed, age, different digestion stimulators, and dietary crude protein (CP) level on the activities of proteolytic enzymes in pancreatic tissue and duodenal chymus (in vivo), serum trypsin and α1-proteinase inhibitor (A1PI) concentrations in meat-type chicks. The study of age dynamics of trypsin and A1PI concentrations was performed on the chicks of hybrid cross “Smena-8”and two parental lines (Plymouth Rock and Cornish) fed standard commercial corn-wheat-SBM diets. Twenty birds per breed were euthanized at 1, 7, 14, 21, 28 and 35 days of age to obtain blood samples and pancreatic homogenate. Experiments on the effects of different digestion promotors (probiotic, acidifier, phytobiotic, enzymatic preparation) and different CP levels (finisher diet, CP 20%, vs. ground corn, CP 8.5%) were performed on 12 hybrid chicks with fistulated duodenum from 14 to 50 days of age. The following conclusions were made: 1) At 1 day of age high proteolytic activity in pancreatic tissue and maximal serum concentrations of trypsin and A1PI were found in both hybrid and parental lines. Since 7 to 35 days of age A1PI concentration was nearly constant, serum trypsin concentration decreased while proteolytic activity in pancreatic tissue exhibited undulate increase;2) Proteolytic activity in pancreatic tissue was higher in hybrids compared to the parental lines from 7 to 35 days of age (p 0.05);3) Supplementation of diet with exogenous enzymes stimulated the digestion due to the increase in protease activity in duodenal chymus by 9.1% compared to unsupplemented control (p 0.05);4) Proteolytic activity in duodenal chymus significantly responded to the substitution of ground corn for the complete diet by 2-fold decrease while serum trypsin concentration responded by 2.5-fold increase (p 0.001). This fact can indicate that physiological functions of digestive proteases are not confined to the digestive processes.

IMMUNOHISTOCHEMICAL STUDY OF ALPHA-1-ANTICHYMOTRYPSIN IN GLIOMA

GFAP is a specific antigen of glial element, but Alpha-1-antichymotrypsin has not been reported in the literature. Alpha-1-antichymotrypsin was guided by GFAP using PAP method to the astrocytes of 137 gliomas. 120 (87%) gliomas were positive for Alpha-1-antichymotrypsin. Of these 120 gliomas, 86 (72%) gave diffuse distribution, 17 (14%) gave focal distribution, and 17 (14%) gave scattered distributions. Alpha-1-antichymotrypsin in glioma tissue may be an important tumor marker for diagnosis.

Managing panniculitis in alpha-1 antitrypsin deficiency: Systematic review of evidence behind treatment

AIM To systematically review literature for management of alpha-1 antitrypsin deficiency(AATD) panniculitis. METHODS Multiple databases were searched using combinations of pertinent terms. Articles were selected describing panniculitis treatment in patients with AAT < 11 μmol and/or PiZZ genotype, with no language limitation. All relevant articles were accessed in full text. Independent review of abstracts and full manuscripts was conducted by 2 reviewers, and quality assessment by one reviewer(checked by a second). Data extraction was conducted byone reviewer(checked by a second). Narrative synthesis only was conducted, as data were unsuitable for metaanalysis.RESULTS Thirty-two case reports and 4 case series were found. Augmentation therapy(infusions of plasma-derived AAT) was the most successful, with complete resolution of symptoms in all patients. Dapsone is a less expensive option, and it achieved clinical resolution in 62% of patients, but it is very poorly tolerated. Among other single-agent antibiotics, doxycycline was the most successful with complete clinical resolution seen in 33% of patients. Immunosuppressants were largely unsuccessful; 80% of patients exhibited no response. Liver transplantation and therapeutic plasma exchange displayed complete resolution in 66% of patients. Other strategies, such as non-steroidal anti-inflammatory drugs or antibiotics other than dapsone did not show sufficient response rates to recommend their use. Authors note the risk of bias imposed by the type of evidence(case reports, case series) available in this field.CONCLUSION Dapsone is the recommended first line therapy for AATD panniculitis, followed by augmentation therapy. Plasma exchange may be an alternative in the setting of rapidly progressive disease.

SlimQuick ^_(TM)-associated hepatotoxicity in a woman with alpha-1 antitrypsin heterozygosity

Green tea (Camellia sinensis)-associated hepatotoxicity is reported. However, the presence of alpha-1 antitrypsin MZ phenotype as a predisposing factor to green tea-associated drug-induced liver injury (DILI) is unknown. A previously healthy woman with alpha-1 antitrypsin MZ phenotype who took SlimQuick?, an herbal supplement containing green tea extract, developed severe hepatotoxicity requiring corticosteroid treatment. Green tea-associated hepatotoxicity is reviewed and alpha-1 antitrypsin MZ phenotype as a predisposing factor to green tea-associated DILI is discussed. Liver biopsy demonstrated marked inflammation with necrosis suggestive of toxic injury with diffuse alpha-1 antitrypsin globule deposition on immunostaining. Corticosteroid therapy resulted in rapid clinical improvement. Alpha-1 antitrypsin MZ phenotype may increase vulnerability to herbal hepatotoxicity.

Alpha-1 Antitrypsin Deficiency Family Study

According to the latest World Health Organization report 64 million people suffer from Chronic Obstructive Pulmonary Disease （COPD）, 3 million people died from COPD and it is predicted that COPD will become the third leading cause of death worldwide by 2030. The alpha-1 antitrypsin deficiency is a rarely diagnosed hereditary disease caused by a genetic mutation and it is one of the most prevalent genetic disorders primarily affecting the lungs, especially in the form of COPD or emphysema, but in some cases also the liver or skin. The Global Initiative for Chronic Obstructive Lung Disease recommends all patients with COPD at a young age or significant family history to be examined for alpha-1 antitrypsin deficiency. This article presents the case of a 42 year old, female patient, Portuguese, with history of Chronic Obstructive Pulmonary Disease, 40 pack units/year smoker, with unknown family history, coming to her family doctor with breath shortness, especially during physical activities, with unsatisfying response to pharmacological prescribed therapy. Physical examination was normal. Alpha- 1 antitrypsin deficiency was confirmed by blood testing. All patient＇s first degree relatives were investigated showing low alpha-1 antitrypsin blood concentrations thus genetic tests were later performed. This case reinforces the need for primary care physicians to be aware of alphal-antitrypsin deficit as an underdiagnosed clinical entity.

alpha-1抗胰蛋白酶在Stanford A型主动脉夹层血管重构中的作用及分子机制的研究

目的:探究α-1抗胰蛋白酶在A型主动脉夹层发病和治疗中的作用与机制。方法:取2012年4月～2016年4月在我院行手术治疗的Stanford A型主动脉夹层患者8例、升主动脉瘤患者8例、正常组8例的资料信息,比较各组血管组织中AAT的蛋白和基因表达水平同时检测血清中AAT含量的变化。使用人原代血管内皮细胞验证AAT对于血管内皮细胞在胰酶刺激下发生细胞凋亡的保护作用,采用皮下缓释angⅡ和血管注射胰酶构建比格犬血管夹层模型,给予AAT治疗后检测Caspase家族蛋白及基因的表达水平。结果:使用RT-PCR及Western blot检测,发现在A型血管夹层患者和主动脉瘤及正常人血管组织中均有不同程度的AAT表达,其中在主动脉瘤患者中AAT表达水平较正常组显著升高(P<0. 05),在A型主动脉夹层患者中AAT表达较正常组显著下调(P<0. 05);在人原代血管内皮细胞中使用AAT预孵育12 h后原代在MMP-2/9压力的作用下,其组织表达的Caspase家族蛋白Caspase-3、Caspase-8较PBS预孵育组显著下调。在动物模型体内,AAT治疗能够显著下调Caspase家族蛋白的mRNA和蛋白水平(P<0. 05)。结论:AAT能够通过抑制Caspase家族蛋白的激活进而抑制血管内皮细胞的凋亡,最终保护血管组织,阻止其形成动脉夹层。

Study of the Effect of Zhuang Medicine Aponeurotic System Triple Therapy on Lumbar Disc Herniation and Alpha-1 Acid Glycoprotein Level

Objective:To analyze the application effect of Zhuang medicine aponeurotic system triple therapy in the treatment of lumbar disc herniation and its effect on the level of alpha-1 acid glycoprotein(alpha-1 AGP).Methods:200 patients with lumbar disc herniation were selected and randomly divided into a treatment group and a control group,100 cases in each group.The control group was given conventional acupuncture,and the treatment group was treated with manipulation+fire needling+cupping.The alpha-1-AGP levels before and after treatment,as well as the lumbar spine function and pain scores before and after treatment,and the adverse reactions occurred during treatment between the two groups were compared.Results:Before treatment,there was no significant difference in alpha-1 AGP levels,lumbar function,and pain scores between the two groups(P>0.05).After treatment,the lumbar function scores of the two groups were significantly increased,with the treatment group having higher scores than the control group(P<0.05);the incidence of adverse reactions in the treatment group was 2.00%,which was much lower than the control group(P>0.05).Conclusion:Appropriate application of Zhuang medicine aponeurotic system triple therapy in the clinical treatment of lumbar disc herniation can promote the improvement of alpha-1 AGP index level,reduce the pain degree of patients,and improve their lumbar spine function.At the same time,Zhuang medicine also has significant advantages in terms of safety,while ensuring the efficacy and safety of the treatment.

Liver Characterization of a Cohort of Alpha-1 Antitrypsin Deficiency Patients with and without Lung Disease

Background and Aims:Alpha-1 antitrypsin deficiency(AATD)is a genetic disorder characterized by the misfolding and accumulation of the mutant variant of alpha-1 antitrypsin(AAT)within hepatocytes,which limits its access to the circulation and exposes the lungs to protease-mediated tissue damage.This results in progressive liver disease secondary to AAT polymerization and accumulation,and chronic obstructive pulmonary disease(COPD)due to deficient levels of AAT within the lungs.Our goal was to characterize the unique effects of COPD secondary to AATD on liver disease and gene expression.Methods:A subcohort of AATD individuals with COPD(n=33)and AATD individuals without COPD(n=14)were evaluated in this study from our previously reported cross-sectional cohort.We used immunohistochemistry to assess the AATD liver phenotype,and RNA sequencing to explore liver transcriptomics.We observed a distinct transcriptomic profile in liver tissues from AATD individuals with COPD compared to those without.Results:A total of 339 genes were differentially expressed.Canonical pathways related to fibrosis,extracellular matrix remodeling,collagen deposition,hepatocellular damage,and inflammation were significantly upregulated in the livers of AATD individuals with COPD.Histopathological analysis also revealed higher levels of fibrosis and hepatocellular damage in these individuals.Conclusions:Our data supports a relationship between the development of COPD and liver disease in AATD and introduces genes and pathways that may play a role in AATD liver disease when COPD is present.We believe addressing lung impairment and airway inflammation may be an approach to managing AATD-related liver disease.

人血清Alpha-1-酸性糖蛋白的糖基化分析

获取真实准确的蛋白质糖基化信息是全面了解糖基化修饰生物学功能的前提.针对简单蛋白质的糖基化分析通常采用反相高效液相色谱-串联质谱技术在肽的水平上对糖基化信息进行采集和解析.本文以人血清Alpha-1-酸性糖蛋白(AGP)酶解液为对象,发展了一套简单有效的蛋白质糖基化分析方法.本方法分为三个步骤,第一步是建立糖肽的理论m/z值表;第二步是获取糖蛋白酶解液的LC-MS谱图,并将每一个色谱峰中所包含糖肽的实际m/z值与理论m/z值进行人工匹配;第三步是对每个色谱峰的糖肽结构归属进行LC-MS/MS验证.采用本方法,我们从AGP酶解液中共鉴定出172条糖肽.与单独采用Survey模式的方法相比,本方法能够显著提高糖肽的覆盖率.

Alpha-bisabolol逆转ABCB1介导的肿瘤多药耐药及机制

目的研究alpha-红没药醇(alpha-bisabolol)是否能有效逆转ATP结合盒转运体B1(ABCB1)介导的肿瘤多药耐药(MDR)并探讨其机制。方法应用网络药理学技术分析仙鹤草(Agrimonia Eupatoria)的活性成分,预测其靶基因,并进行基因本体论(GO)与京都基因与基因组百科全书(KEGG)通路分析。进一步通过分子对接及热稳定性实验分析alpha-bisabolol和ABCB1的互作。通过MTT法检测alpha-bisabolol能否有效逆转ABCB1介导的肿瘤MDR。通过Western blot法、免疫荧光与流式细胞术研究alpha-bisabolol逆转ABCB1介导肿瘤MDR的机制。结果网络药理学实验结果表明,Agrimonia Eupatoria的活性成分alpha-bisabolol与肿瘤的发生密切相关。分子对接及热稳定性实验结果显示,alpha-bisabolol能够与ABCB1形成稳定的结合。逆转实验结果表明,alpha-bisabolol能够有效逆转ABCB1介导的肿瘤MDR。此外,机制实验结果显示,alpha-bisabolol对肿瘤细胞内ABCB1蛋白的表达及定位并不产生影响。alpha-bisabolol通过抑制ABCB1蛋白的外排功能,提高了细胞内药物的蓄积水平,从而有效逆转肿瘤MDR。结论alpha-bisabolol能够有效逆转ABCB1介导的肿瘤MDR,为化疗耐药的肿瘤患者提供了一种新的治疗策略。

Thymosin Alpha-1 Inhibits Complete Freund’s Adjuvant-Induced Pain and Production of Microglia-Mediated Pro-inflammatory Cytokines in Spinal Cord

Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been reported to have anti-inflammatory and neuroprotective functions in rodents.However,how Tα1 affects inflammatory pain remains unclear.In the present study,intraperitoneal injection of Tal attenuated complete Freund's adjuvant(CFA)-induced pain hypersensitivity,and decreased the up-regulation of pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6)in inflamed skin and the spinal cord.We found that CFA-induced peripheral inflammation evoked strong microglial activation,but the effect was reversed by Tα1.Notably,Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter(VGLUT)and down-regulated the vesicular γ-aminobutyric acid transporter(VGAT)in the spinal cord.Taken together,these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microgliainduced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.

拟南芥Alpha-dioxygenase1的原核表达、纯化及活性的检测

从经水杨酸处理的拟南芥开花期植株中获得cDNA,扩增得到Alpha-dioxygenase 1(DOX1)基因,进行原核表达、纯化和生物活性检测。利用原核表达载体pMAL-c4x在T7 Express CompetentE.coli和BL21(DE3)-RIPL codon+菌株中表达DOX1,经Amylose Resin亲和层析柱纯化。SDS-PAGE结果表明,重组融合蛋白在BL21(DE3)-RIPL codon+中的表达通过灰度值比较分析以可溶性为主,表达率约3.7%,纯化的DOX1纯度可达45%。愈创木酚法表明,可溶性重组蛋白不具有过氧化物酶活性;2,4-DNP法测试表明可溶性重组蛋白具有脂肪酸双加氧酶活性。

Associations of Polymorphism of rs9944155, rs1051052, and rs1243166 Locus Allele in Alpha-1-antitrypsin with Chronic Obstructive Pulmonary Disease in Uygur Population of Kashgar Region

Background： Previous studies conducted in various geographical and ethnical populations have shown that Alpha-1 -antitrypsin （Alpha-1-AT） expression affects the occurrence and progression of chronic obstructive puh-nonary disease （COPD）. We aimed to explore the associations of rs9944155AG, rsl051052AG, and rs1243166AG polymorphisms in the A lpha-1-A T gene with the risk of COPD in Uygur population in the Kashgar region. Methods： From March 2013 to December 2015. a total of 225 Uygur COPD patients and 198 healthy people were recruited as cases and controls, respectively, in Kashgar region. DNA was extracted according to the protocol of the DNA genome kit, and Sequenom MassARRAY single-nucleotide polymorphism technology was used for genotype determination. Serum concentration of Alpha-1-AT was detected by enzyme-linked immunosorbent assay. A logistic regression model was used to estimate the associations of polymorphisms with COPD. Results： The rs1243166-G allele was associated with a higher risk of COPD （odds ratio [OR] = 2.039, 95% confidence interval [CI]： 1.116-3.725, P = 0.019）. In cases, Alpha-1-AT levels were the highest among participants can-yiug rs1243166 AG genotype, followed by AA and GG genotype （χ2 = 11.89, P = 0.003）. Similarly, the rs1051052-G allele was associated with a higher risk of COPD （OR = 19.433, 95% CI： 8.783-43.00, P 〈 0.001）. The highest Alpha-1-ATlevels were observed in cases carrying rs1051052 AA genotype, followed by cases with AG and GG genotypes （χ2= 122.45, P 〈 0.001）. However, individuals with rs9944155-G allele exhibited a lower risk of COPD than those carrying the rs9944155-A allele （OR = 0.121, 95% CI： 0.070-0.209, P 〈 0.001 ）. in both cases and controls, no significant difference in Alpha-l-AT levels was observed among various rs9944115 genotypes. Conclusions： rs 1243166, rs9944155, and rs 1051052 sites of Alpha- I-A Tmay be associated with the COPD morbidity in Uygur population. While rs 1243166-G allele and rs1051052-G allele are associated with an increased risk of developing COPD, rs9944155-G allele is a protect locus in Uygur population. Alpha1-AT levels in Uygur COPD patients were lower than those in healthy people and differed among patients with different rs 1051052 AG and rs 1243166 AG genotypes.

原发性肺腺癌alpha-B-crystallin、TTF-1、NapsinA表达研究

目的分析alpha-B-crystallin、甲状腺转录因子1(TTF-1)、Napsin A在原发性肺腺癌中的表达及意义,为今后该病临床诊断及治疗提供参考。方法收集原发性肺腺癌患者80例作为观察组,将鳞癌、腺癌、腺鳞癌、大细胞癌(共50例)归为对照组;使用免疫组化检测alpha-B-crystallin、TTF-1、Napsin A,分析三指标与原发性肺腺癌病例类型、分期、预后等相关性。结果 alpha-B-crystallin、TTF-1、Napsin A在观察组(肺腺癌)中阳性率分别为20.00%、81.25%、88.75%,与对照组有显著差异(P<0.05);Napsin A、TTF-1、alpha-B-crystallin实性为主阳性率分别为54.20%、65.03%、0;alpha-B-crystallin阳性率在不同肿瘤大小、分期、预后中有显著差异;Napsin A阳性率在不同肿瘤大小、分期、组织分化、淋巴结转移、预后中有显著差异;TTF-1阳性率在不同肿瘤大小、组织分化、分期、预后中有显著差异(P均<0.05)。结论 1TTF-1、Napsin A阳性表达能明确区分原发性肺腺癌、鳞癌、大细胞癌和腺鳞癌,可辅助临床肺癌鉴别;2TTF-1、Napsin A在原发性肺腺癌中有高阳性率,用于肺腺上皮肿瘤病理诊断能提高肺腺癌诊断准确性;3alpha-B-crystallin、TTF-1、Napsin A阳性表达均与肺腺癌病例分期、肿瘤大小、预后相关,对该病预后判断有辅助作用。