Effect of viral load on T-lymphocyte failure in patients with chronic hepatitis B

AIM: To investigate peripheral T-lymphocyte sub-population profile and its correlation with hepatitis B virus (HBV) replication in patients with chronic hepatitis B (CHB).METHODS: Distribution of T-lymphocyte subpopulations in peripheral blood was measured by flow cytometry in 206 CHB patients. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time polymerase chain reaction (PCR). The relationship between HBV replication and variation in peripheral T-cell subsets was analyzed.RESULTS: CHB patients had significantly decreased CD3+ and CD4+ cells and CD4+/CD8+ ratio, and increased CD8+ cells compared with uninfected controls (55.44 ± 12.39 vs 71.07 ± 4.76, 30.92 ± 7.48 vs 38.94 ± 3.39, 1.01 ± 0.49 vs 1.67 ± 0.33, and 34.39 ± 9.22 vs 24.02 ± 4.35; P < 0.001, respectively). Univariate analysis showed a similar pattern of these parameters was significantly associated with high viral load, presence of serum hepatitis B e antigen (HBeAg) expression, liver disease severity, history of maternal HBV infection, and young age at HBV infection, all with P < 0.01. There was a significant linear relationship between viral load and these parameters of T-lymphocyte subpopulations (linear trend test P < 0.001). There was a negative correlation between the levels of CD3+ and CD4+ cells and CD4+/CD8+ ratio and serum level of viral load in CHB patients (r = -0.68, -0.65 and -0.75, all P < 0.0001), and a positive correlation between CD8+ cells and viral load (r = 0.70, P < 0.0001). There was a significant decreasing trend in CD3+ and CD4+ cells and CD4+/CD8+ ratio with increasing severity of hepatocyte damage and decreasing age at HBV infection (linear trend test P < 0.01). In multiple regression (after adjustment for age at HBV infection, maternal HBV infection status and hepatocyte damage severity) log copies of HBV DNA maintained a highly significant predictive coefficient on T-lymphocyte subpopulations, and was the strongest predictor of variation in CD3+, CD4+, CD8+ cells and CD4+/CD8+ ratio. However, the effect of HBeAg was not significant.CONCLUSION: T-lymphocyte failure was signifi-cantly associated with viral replication level. The substantial linear dose-response relationship and strong independent predictive effect of viral load on T-lymphocyte subpopulations suggests the possibility of a causal relationship between them, and indicates the importance of viral load in the pathogenesis of T cell hyporesponsiveness in these patients.

Hepatitis B virus DNA is more powerful than HBeAg in predicting peripheral T-lymphocyte subpopulations in chronic HBV-infected individuals with normal liver function tests

AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.

Cytotoxic T-lymphocyte antigen 4 gene polymorphisms and susceptibility to chronic hepatitis B

AIM： To assess the three polymorphJsm regions within cytotoxic T-lymphocyte antigen 4 （CTLA-4） gene, a C/T base exchange in the promoter region-318 （CTLA-4 -318C/T）, an A/G substitution in the exon 1 position 49 （CTLA-4 49A/G）, a T/C substitution in 1172 （CTLA-4 -1172T/C） in patients with chronic hepatitis B. METHODS： Fifty-one patients with chronic hepatitis B virus infection and 150 healthy subjects were recruited sequentially as they presented to the hepatic clinic. Classification of chronic hepatitis B virus （HBV）-infected patients was as asymptomatic carrier state （26 patients） and chronic hepatitis B （25 patients）. Genomic DNA was isolated from anti-coagulated peripheral blood Bully coat using Miller＇s salting-out method. The presence of the CTLA-4 gene polymorphisms was determined using polymerase chain reaction amplification refractory mutation system （ARMS）. RESULTS： We observed a significant association between -318 genotypes frequency （T＋C-, T＋C＋, T-C＋） and susceptibility to chronic hepatitis B （P=0.012, OR=0.49, 95%CI： 0.206-1.162）. However, we did not observe a significant association for ＋49 genotype frequency （T＋C＋, T＋C- T-C＋） and -1172 genotype frequency （C＋T＋, T＋C- C＋T-） and state of disease. CONCLUSION： Our results suggest that CTLA-4 gene polymorphisms may partially be involved in the susceptibility to chronic hepatitis B.

High value of controlled attenuation parameter predicts a poor antiviral response in patients with chronic hepatits B

BACKGROUND: Controlled attenuation parameter (CAP) is a non-invasive method for diagnosing hepatic steatosis based on vibration-controlled transient elastography. The objective of this study was to investigate the effect of high value of CAP on antiviral therapy in patients with chronic hepatitis B (CHB). METHODS: Patients with CHB receiving enticavir for initial antiviral therapy were studied; they were divided into the high CAP group and normal CAP group at baseline according to the CAP values. The effect of the antiviral therapy between the two groups were compared at week 12, 24 and 48. Patients with high CAP value at baseline were divided into three subgroups, mild, moderate and severe elevation; the therapeutic response were compared among patients with normal CAP and subgroups of patients with elevated CAP. RESULTS: A total of 153 patients were enrolled. Among them, 63 were in the high CAP group and 90 in the normal CAP group. Patients with high CAP had lower rates of ALT normalization and HBV DNA clearance in response to antiviral therapy compared with those with normal CAP at week 12, 24 and 48. Further analysis showed that the rate of ALT normalization in patients with mildly and moderately elevated CAP were significant lower than those with normal CAP at week 12 and 24; while the difference was not significant between the patients with normal CAP and those with severely elevated CAP. The rate of HBV DNA clearance was significantly lower in patients with severely elevated CAP compared with those with normal CAP at week 12, 24 and 48. CONCLUSION: CHB patients with high CAP had poor response to antiviral therapy.

Oral Immunization of Mice With Vaccine of Attenuated Salmonella typhimurium Expressing Helicobacter pylori Urease B Subunit

Objective To prepare the live recombinant vaccine of attenuated Salmonella typhimurium SL3261 expressing Helicobacterpylori （H. pylori） B subunit （UreB） and to determine whether it could be used as an oral vaccine against H. pylori infection. Methods Using genomic DNA of H. pylori Sydney strain （SS1） as template, the H. pylori UreB gene fragment was amplified by PCR and subcloned into the expression vector pTC01. The recombinant plasmid pTC01-UreB was then transferred into LBS000 to obtain modified forms, and further conversed into the attenuated Salmonella typhimurium SL3261 to obtain recombinant SL3261/pCT01-UreB as an oral immunization reagent, which was then used to orally immunize Balb/c mice twice at a three-week interval. Twelve weeks later, anti-UreB IgA antibodies in intestinal fluid and IgG antibodies in sera were determined by ELISA. The relating data in control groups （including body weight, gastric inflammation, etc.） were also collected. Results The sequencing analysis showed that the UreB gene fragment amplified by PCR was consistent with the sequence of the H. pylori UreB gene. The restriction enzyme digestion revealed that the correct pTC01-UreB was obtained. SDS-PAGE and Western blot showed that a 61KD protein was expressed in SL3261/pTC01-UreB, which could be recognized by anti-H, pylori UreB antiserum and was absent in the control containing only Salmonella typhimurium SL3261 strain. The multiple oral immunization with SL3261/pTC01-UreB could significantly induce H. pylori specific mucosal IgA response as well as serum IgG responses. IFN-T and IL-10 levels were significantly increased in SL3261/pTC01-UreB group, and no obvious side effect and change in gastric inflammation were observed. Conclusion The attenuated vaccine of Salmonella typhimurium expressing H. pylori UreB can be used as an oral vaccine against H. pylori infection.

Immunogenicity and protective efficacy of DHBV DNA vaccines expressing envelope and capsid fusion proteins in ducks delivered by attenuated Salmonella typhimurium

Duck hepatitis B virus(DHBV) shares many basic characteristics with hepatitis B virus(HBV) and is an attractive model for vaccine development. In this study, DHBV DNA vaccines were designed to express envelope and capsid fusion proteins to enhance the breadth of immune response in ducks. Attenuated Salmonella typhimurium(SL7207) was used as a carrier and adjuvant to boost the magnitude of immune response. Based on this strategy, novel DNA vaccines(SL7207-p VAX1-LC and SL7207-p VAX1-SC) were generated. Growth kinetics, genetic stabilities and relative transcription levels of the L, S and C genes introduced by these vaccine strains were measured before inoculation to guarantee safety and efficacy. The relative transcript levels of the CD4 and CD8 T genes and the antibody levels(Ig Y) in ducks receiving the vaccines were higher than those in single gene delivered groups. Additionally, the copy number of covalently closed circular DNA in hepatocytes after DHBV challenge also provided evidence that our fusion vaccines could enhance the protective efficiency against DHBV infection in ducks.

Diagnostic value of controlled attenuation parameter for liver steatosis in patients with chronic hepatitis B

AIM: To study the diagnostic value of controlled attenuation parameter (CAP), evaluated by transient elastography, for liver steatosis in patients with chronic hepatitis B (CHB).

B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价

旨在研究B亚型禽偏肺病毒减毒株(LN16-A株)对商品肉鸡的免疫效果。本研究从规模化养殖场选取3周龄的商品肉鸡,将B亚型禽偏肺病毒减毒株(LN16-A株)通过滴鼻方式免疫(200μL,病毒含量≥103 TCID50·只^(-1)),空白对照组以同样剂量的PBS滴鼻。免疫21 d后,翅下采集血液分离血清,利用ELISA方法及中和试验检测相应抗体;在6周龄时,通过滴鼻方式使用B亚型禽偏肺病毒强毒(LN16-V株)(200μL,5000 TCID50·只-1)进行攻毒,攻毒后连续观察7 d,并利用RT-qPCR检测各组鸡鼻腔拭子的病毒拷贝数,攻毒后第9天各组随机选取2只鸡进行剖杀,采集鼻甲骨、气管和肺组织,通过HE染色制作病理切片,观察组织病理学变化。抗体检测结果显示,在疫苗免疫后21 d,免疫组ELISA抗体和中和抗体阳性率均为100%,平均ELISA抗体效价为1860,平均中和抗体效价为7.44 log2;临床症状观察结果显示,对照组商品肉鸡出现混浊、黏稠、牵丝状的鼻液等明显临床症状,发病率为100%,而免疫组肉鸡无临床症状;RT-qPCR结果表明,免疫组鸡鼻腔拭子病毒基因组拷贝数相较于对照组降低了78%~96%;病理切片结果显示,对照组肉鸡鼻甲骨、气管以及肺均出现不同程度的病理损伤,免疫组肉鸡各组织器官未见明显病理变化。综上表明,B亚型禽偏肺病毒减毒株(LN16-A株)可以诱导商品肉鸡产生特异性抗体,能显著降低攻毒后的排毒水平,对商品肉鸡具有良好的保护效果。本研究结果将为肉鸡养殖场禽偏肺病毒病的防控提供重要理论指导和技术支撑。

茯苓甘草合剂辅助西药对慢性阻塞性肺疾病合并肌肉衰减患者NF-κB信号通路的影响

目的探讨茯苓甘草合剂辅助西药对慢性阻塞性肺疾病(COPD)合并肌肉衰减NF-κB信号通路的影响及分子机制。方法选取2020年8月至2022年8月COPD合并肌肉衰减患者104例,作为研究对象随机数字表法分为对照组和观察组,每组52例。对照组给予西药治疗,观察组给予西药+茯苓甘草合剂治疗。统计2组中医疗效、不良反应及治疗前后中医证候积分、NF-κB mRNA、蛋白及因子[白细胞介素-8(IL-8)、白细胞介素-1(IL-1)、肿瘤坏死因子-α(TNF-α)]、营养状况指标[转铁蛋白(Tf)、血红蛋白(Hb)、白蛋白(ALB)]、肌肉衰减相关指标[握力试验、起立-行走计时测试(TUGT)、5次坐立试验(FTSST)、肌少症筛查问卷(SARC-F)]。结果观察组中医治疗总有效率[86.54%(45/52)]高于对照组[67.31%(35/52)](P<0.05);治疗1个月、3个月后观察组中医证候积分低于对照组(P<0.05);治疗1个月、3个月后观察组NF-κB mRNA、NF-κB蛋白、IL-8、IL-1、TNF-α低于对照组(P<0.05);治疗3个月后观察组握力高于对照组,FTSST、TUGT、SARC-F评分低于对照组(P<0.05);治疗期间,2组均未出现明显不良反应。结论茯苓甘草合剂辅助西药治疗COPD合并肌肉衰减效果确切,可通过抑制NF-κB信号通路,减轻炎性反应,改善患者营养状态,有效改善临床症状,且安全性高。

新风胶囊对膝骨关节炎患者B、T淋巴细胞衰减因子及氧化应激的影响

目的探讨膝骨关节炎患者(knee osteoarthritis,KOA)B、T淋巴细胞衰减因子(B and T lymphocyte attenuator,BTLA)及超氧化物歧化酶(superoxide dismutase,SOD)的变化及新风胶囊(XinFeng capsule,XFC)对其影响。方法将60例KOA患者按随机分为2组:30例治疗组,即XFC组(每次3粒,每日3次)和30例对照组,即氨基葡萄糖组(glucosamine,GS组,每次2粒,每日3次);2组治疗均30天为1个疗程,连续1个疗程。流式细胞术检测BTLA表达频率及活化水平,比色法测定血清SOD,采用魏氏法测定血沉(erythrocyte sedimentation rate,ESR),全自动生化分析仪测定hs-C反应蛋白(hypersensitive cross-reacting protein,hs-CRP)。酶联免疫吸附法检测2组血清中白介素(interleukin,IL)-10,IL-1β,丙二醛(methane dicarboxylic aldehyde,MDA)。结果①与NC组比较,KOA患者外周血BTLA在CD3+T细胞(CD3+BTLA+T cell)、CD8+T细胞(CD8+BTLA+T cell),CD4+T细胞上(CD4+BTLA+Tcell)的表达频率明显下降,MAD明显升高。与正常范围比较,60%的KOA患者血清SOD水平降低,且随Lequesne MG的加重及病程的延长而降低,而与年龄、性别无关。②Pearson相关分析结果显示:外周血BTLA表达水平与Lequesne MG呈明显负相关,与RP、RE、BP积分呈正相关;CD3+BTLA+T cell、CD4+BTLA+T cell与IL-1β呈明显负相关,与血清IL-10呈正相关;CD4+BTLA+T cell与血清SOD呈正相关(P<0.05或P<0.01)。③与对照组相比,SF-36总积分、BTLA表达频率、IL-10明显升高,Lequesne MG、症状分级量化评分、IL-1β、MDA明显降低。结论 XFC能提高KOA患者外周血BTLA表达,抑制T细胞活化,降低异常免疫反应和氧化应激损伤,减轻关节疼痛症状,改善患者全身机能,从而提高生活质量水平。

四种风湿病患者血小板参数变化及外周血BTLA、Treg观察

目的:观察四种风湿病患者血小板参数变化及外周血B、T淋巴细胞衰减因子(BTLA)、调节性T细胞(Treg)的表达。方法:采用流式细胞术检测风湿病患者外周血BTLA、Treg表达,生化仪检测血清血小板参数及其他实验室指标,观察风湿病患者血小板参数变化及与BTLA、Treg等的关系。结果:四种风湿病患者血小板异常率最高为RA,其次为AS、SS、OA;与正常组比较,这四种风湿病患者外周血BTLA、Treg表达降低;BTLA、Treg异常组血小板计数(PLT)、血小板压积(PCT)、血小板分布宽度(PDW)较BTLA、Treg正常组升高(P<0.05或P<0.01);血小板参数与BTLA、Treg呈相关性。结论:四种风湿病患者血小板异常可能与BTLA、Treg表达失衡有关。

新风胶囊治疗对强直性脊柱炎患者BTLA^+T细胞数量和氧化应激的影响

目的探讨强直性脊柱炎(AS)患者B、T淋巴细胞衰减因子(BTLA)及活性氧(ROS)、活性氮(RNS)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、总抗氧化能力(TAOC)的变化及新风胶囊(XFC)对其影响。方法将140例AS患者随机等分为XFC组(3粒/次,tid)和柳氮磺吡啶(SASP)组(4片/次,bid);连续治疗3个月。60例健康体检者为对照组。采用流式细胞术检测患者外周血BTLA表达频率及活化水平;采用ELISA检测2组血清中氧化应激指标(ROS、RNS、MDA、SOD、CAT、TAOC)和白细胞介素4(IL-4)、IL-10、IL-1β、肿瘤坏死因子α(TNF-α)含量;采用魏氏法检测血沉(ESR);采用日立7060型全自动生化分析仪检测高敏C反应蛋白(Hs-CRP)。结果 XFC组临床疗效显著优于SASP组,具有统计学意义(P<0.01)。与健康对照组相比,AS患者外周血CD3+T细胞、CD4+T细胞BTLA的水平显著降低(P<0.01或P<0.05);与对照组相比,AS患者SOD、CAT、TAOC值均显著降低,ROS、RNS、MDA值显著升高(P<0.01或P<0.05);与对照组相比,AS患者血清IL-1β、TNF-α和ESR、Hs-CRP值显著升高(P<0.01),IL-4、IL-10水平显著降低(P<0.01或P<0.05)。与治疗前相比,2组治疗后外周血BTLA+CD3+T细胞、BTLA+CD4+T细胞、SOD、TAOC、IL-4,SF-36量表生理机能(PF)、社会功能(SF)、生理职能(RP)、情感职能(RE)、躯体疼痛(BP)、精神健康(MH)、活力(VT)以及一般健康状况(GH)八个维度积分均显著升高,ROS、MDA、TNF-α、ESR、Hs-CRP、VAS、BASDAI、BASFI、BAS-G均显著降低(P<0.01或P<0.05)。XFC组与SASP组相比,差异有统计学意义(P<0.01或P<0.05)。Pearson相关分析结果显示:外周血BTLA表达水平与SOD、RP、BP、SF、RE呈正相关;BTLA+CD3+T细胞、BTLA+CD4+T细胞与ROS、MDA、IL-1β、TNF-α、ESR、VAS、BASDAI呈明显负相关,与TAOC、IL-4、IL-10呈正相关;BTLA+CD3+T细胞与RNS、Hs-CRP、BASFI呈明显负相关;BTLA+CD4+T细胞与CAT呈正相关。结论 XFC能增强AS患者外周血BTLA表达,负性调节T细胞的激活与增殖。

BTLA在重症社区获得性肺炎患者中的表达及糖皮质激素干预动物模型的影响

目的本研究通过观察BT淋巴衰减子(B and T lymphocyte attenuator,BTLA)在重症CAP患者外周血淋巴细胞及支气管黏膜上的表达,外源性干预LPS-诱导的急性肺部炎症小鼠模型BTLA表达对肺部炎症的作用以及给予糖皮质激素对BTLA表达的影响。方法利用流式细胞学分析重症CAP患者外周血BTLA^+CD4^+淋巴细胞比例,另外对重症CAP患者支气管黏膜常规行HE染色和免疫组织化学检测BTLA表达情况。利用流式细胞学分析LPS诱导的急性肺部炎症模型右心血BTLA^+CD4^+淋巴细胞比例。收集肺泡灌洗液(bronchoalveolar lavage fluid,BALF)行炎症细胞分类计数,并利用ELISA检测炎症因子。取肺组织分别行HE染色、免疫组织化学染色和蛋白印迹。结果流式细胞学分析重症肺炎患者外周血中BTLA^+CD4^+淋巴细胞比例明显高于正常对照组。此外,支气管黏膜HE染色结果显示重症CAP患者支气管黏膜有大量炎症细胞浸润,并且伴有支气管壁增厚;进一步用免疫组织化学检测支气管黏膜BTLA表达,结果提示重症肺炎患者支气管黏膜BTLA表达明显增高,浸润的炎症细胞BTLA表达增高明显。流式细胞学分析LPS诱导的急性肺部炎症模型小鼠右心血中BTLA^+CD4^+淋巴细胞比例明显高于正常对照组。利用激动性抗-BTLA抗体6A6干预急性肺部炎症模型后淋巴细胞BTLA表达较模型小鼠明显增高,同时,激动性抗-BTLA抗体6A6能减轻肺部炎症反应和抑制NF-κB信号通路激活。利用糖皮质激素干预急性肺部炎症模型后淋巴细胞BTLA表达较模型小鼠明显增高,糖皮质激素能减轻肺部炎症反应和抑制NF-κB信号通路激活。结论通过本研究我们推测BTLA信号可能参与了重症CAP患者的免疫调节过程,BTLA可能成为重症CAP治疗的一个新靶点。同时,BTLA的表达可能反应患者机体的免疫状态,为临床使用糖皮质激素治疗提供指导作用。

携带HBV包膜大蛋白DNA疫苗的减毒沙门菌在小鼠诱导免疫应答

探索一种简便、有效的乙型肝炎病毒DNA疫苗免疫方法。将编码绿色荧光蛋白的真核表达质粒pEGFPN1转化到减毒鼠伤寒沙门菌SL72 0 7,灌胃饲服BALB c小鼠 ,流式细胞术检测出小鼠脾细胞内表达的绿色荧光蛋白 ;构建编码HBV包膜大蛋白的DNA疫苗pCI S1S2S ,分别以SL72 0 7为载体的口服途径或直接肌肉注射途径免疫BALB c小鼠 ,检测小鼠的血清抗体、T细胞增殖和细胞毒性T淋巴细胞反应 ,结果表明两种免疫途径均能在小鼠体内诱生细胞和体液免疫应答 ,但口服途径诱导免疫应答的强度明显强于肌肉注射途径。口服携带HBVDNA疫苗的减毒伤寒沙门菌可能代表一种简便。

风湿病患者肺功能降低与BTLA阳性细胞及调节性T细胞的数量减少有关

目的探讨B、T淋巴细胞衰减因子(BTLA)、调节性T细胞(Treg)与风湿病患者肺功能降低的关系。方法选取482例风湿病患者,包括类风湿性关节炎(RA)198例,强直性脊柱炎(AS)114例、干燥综合征(SS)102例、骨关节炎(OA)68例,应用肺功能仪检测患者肺功能水平,采用流式细胞术检测患者外周血B、T淋巴细胞衰减因子(BTLA)和调节性T细胞(Treg)表达。结果与正常组比较,风湿病患者肺功能降低,外周血BTLA、Treg表达降低;与风湿病BTLA、Treg正常组比较,BTLA、Treg异常组肺功能参数第1秒用力呼气容积(FEV1)、最大呼气流量(PEF)、50%肺活量位的最大呼气流量(FEF50)、FEF75降低(P<0.05或P<0.01);相关分析显示,肺功能参数用力肺活量(FVC)、最大通气量(MVV)、FEV1、FEF25、FEF50分别与BTLA、Treg呈正相关,FVC、FEV1、FEF50、FEF75分别与IgA、IgM呈负相关(P<0.05或P<0.01)。结论风湿病患者肺功能降低可能与BTLA、Treg表达降低,促使T细胞、B细胞过度异常活化有关。

类风湿关节炎患者心肺功能变化及与B、T淋巴细胞衰减因子及氧化应激的相关性分析

目的观察类风湿关节炎患者的心、肺功能变化及其与外周血BTLA和氧化应激指标变化的相关性,探讨心肺功能下降的机制。方法选取RA患者100例并抽取40例健康人作为正常对照。采用肺功能仪检测患者肺功能参数FVC、FEV1、MVV、PEF、MEF25、MEF50、MEF75。采用超声心动图检测2组心功能参数EF%、SV%、FS%、E、A、E/A。流式细胞术检测BTLA表达频率及活化水平,魏氏法测定ESR,全自动生化仪测定hs-CRP。酶联免疫吸附法检测外周血细胞因子(IL-17、TNF-α、IL-4、IL-35)及氧化应激指标(MDA、ROS、SOD、TAOC)。结果 1)与NC组相比,RA患者肺功能参数均有不同程度降低;心功能参数EF、E峰、E/A明显降低,A峰明显升高,SV无明显差异;外周血IL-17、TNF-α和炎性指标ESR、CRP值显著升高,BTLA、IL-4、IL-35值显著降低,ROS、MDA明显升高,SOD、TAOC明显降低。2)相关性分析显示心功能参数与CD24+细胞、CD19+CD24+细胞、TAOC负相关,与BTLA、CD19+Cell、ROS、MDA、SOD呈正相关;肺功能各参数均与ESR、Hs-CRP、DAS28呈负相关,与BTLA、CD19+CD24+、SOD、TAOC、IL-4、IL-35呈正相关,与CD24+BTLA+、ROS、MDA、IL-17、TNF-α呈负相关。结论 RA患者存在心肺功能降低,并与机体BTLA表达减弱、抗氧化能力下降密切相关。

滤减UV-B辐射对烟叶可溶性蛋白、光合色素和类黄酮的影响

在云南玉溪烟区种植烤烟海拔最高(1806.0m)的通海县,通过盆栽烤烟K326试验,研究了在滤减自然的太阳UV-B辐射强度25%、50%和65%条件下,UV-B辐射对烟叶发育过程中可溶性蛋白、光合色素和类黄酮的影响。结果表明:随叶龄增加,可溶性蛋白含量下降,光合色素降解,类黄酮在老叶中积累,蛋白质在生理成熟期对UV-B辐射最敏感。与对照相比,减弱UV-B辐射处理降低了烟叶类黄酮和可溶性蛋白含量,但光合色素含量上升;较低的UV-B辐射降低了叶绿素的降解速度。结果从一侧面说明UV-B辐射对烟叶蛋白质的合成是有益的,类黄酮和叶绿素的变化是对UV-B辐射变化的适应性反应,类黄酮与蛋白质之间可能存在一定的偶联关系。

两种栽培荞麦对日光UV-B辐射的响应

两种荞麦(甜荞和苦荞)栽培在不同紫外线B(UV-B)辐射[减弱(-UVB)、近充足(NA,Near-abient)、增强(+UVB)]保护小区内,以研究当前和未来增强UV-B辐射对其生长、发育、产量、生物量和卢丁含量等的影响。结果表明,不仅未来增强UV-B辐射而且当前高原条件下的UV-B辐射显著抑制了荞麦的生长、降低了产量和生物量积累,但促进了荞麦发育使其开花期提前。另一方面,叶片卢丁和UV吸收化合物在NA和增强UV-B辐射下显著增加,而多酚氧化酶在UV-B辐射下大幅度提高。两种栽培荞麦对UV-B辐射非常敏感,在高紫外辐射地区或未来增强UV-B辐射条件下荞麦生产除应考虑到高产外,还应该考虑到荞麦对紫外线辐射的耐性。

低纬高原滤减UV-B辐射对烤烟营养生长期形态性状的影响

在云南玉溪主产烟区的通海县(海拔1806.0m)自然环境中,通过覆膜减弱UV-B的辐射强度(分别是自然环境下的25%、50%和65%),采用盆栽试验研究了UV-B辐射对烤烟K326团棵中后期-现蕾期部分形态特征的影响。结果表明:减弱自然环境下的UV-B辐射对K326株高、最大叶长、叶宽及叶面积、茎围、节间距和叶数等没有显著影响。但在不同减弱UV-B处理下,株高、最大叶长、最大叶面积和节间距等呈现增加的趋势,而叶数则有减少的趋势。说明目前通海的UV-B辐射水平已对K326的形态建成产生了一定的抑制效应。

烤烟叶片光合气体交换参数对滤减UV-B辐射强度的响应

以烤烟品种‘K326’为试验材料,通过覆盖不同的透明薄膜滤减UV-B辐射处理方式,并应用多元统计的岭回归分析方法,研究了100%(CK)、75%(A)、50%(B)、35%(C)自然UV-B辐射处理下烤烟成熟初期叶片的光合气体交换参数与50 cm和150 cm高度UV-B辐射强度的关系。结果表明:对于净光合速率,不同高度的UV-B辐射对A、B、C处理的抑制强度都是A>C>B;3个处理的瞬时水分利用效率(WUE)受UV-B辐射抑制强度为B>C>A;在A、C处理条件下,影响烟叶净光合速率变化的主要是气孔因素,而B处理条件下却主要是非气孔因素。可见,‘K326’烤烟叶片的光合气体交换参数对不同的UV-B辐射强度响应区间的敏感性存在一定的差异。