目的 通过分析HBV表面抗原(HBsAg)的表达量与乙型肝炎病毒DNA(HBV DNA)、乙型肝炎e抗原(HBeAg)及丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)的关系,探讨HBsAg表达量在慢性乙型病毒性肝炎(慢乙肝)患者诊疗中的意义.方法 收集2012年12月—2015年12月在石家庄市第五医院就诊的慢乙肝患者624例,采用酶免疫发光技术检测血清HBsAg及HBeAg水平,根据HBsAg值将患者分为0～50 IU/mL组、50～1000 IU/mL组、1000～5000 IU/mL组、5000～10000 IU/mL组、10000～25000 IU/mL组及>25000 IU/mL组,采用荧光定量PCR技术检测各组HBV DNA表达量,采用全自动生化仪检测各组血清ALT、AST水平,分析HBsAg不同定量值的临床意义.结果 随着患者血清中HBsAg水平不断增高,HBV DNA表达量及HBeAg阳性率亦显示增高趋势,10000～25000 IU/mL组与>25000 IU/mL组的HBV DNA表达量及HBeAg阳性率比0～50 IU/mL组、50～1000 IU/mL组、1000～5000 IU/mL组、5000～10000 IU/mL组明显增高,差异均有统计学意义(P均<0.05).HBeAg阳性患者血清HBsAg水平比HBeAg阴性患者明显增高(P<0.05).各组ALT、AST水平呈异常状态,只有HBsAg>25000 IU/mL组的患者ALT、AST水平多数在正常范围.结论 随着HBsAg水平不断增高,患者体内HBV复制活跃,且病毒数量不断增多;而HBsAg高于25000 IU/mL的患者多数处于免疫耐受期,位于1000～5000 IU/mL的患者多数处于低复制期,这部分患者均需密切监视,有利于疾病及时治疗.

目的 探讨干扰素α-1b序贯拉米夫定联合治疗对慢性乙型肝炎患儿T淋巴细胞亚群、补体表达水平的影响,并对其肝功能进行评价.方法 选取2015年11月～2017年12月石家庄市第五医院(以下简称“我院”)收治的136例慢性乙型肝炎患儿作为研究对象,根据随机数字表法将其分为研究组和对照组,每组68例.对照组患者仅给予肌内注射干扰素α-1b.研究组采用拉米夫定联合干扰素α-1b治疗.观察两组治疗前后T淋巴细胞亚群(CD4+、CD8+、CD4+/CD8+)及CD8+T细胞PD-1、Tim-3表达量、补体(C3、C4)水平;观察两组治疗前后谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBil)水平的变化.结果 研究组治疗后CD4+、CD4+/CD8+水平高于对照组,CD8+水平低于对照组,差异均有统计学意义(P＜0.05).研究组治疗后CD8+T细胞PD-1、CD8+ Tim-3水平低于对照组,C3、C4水平均高于对照组,差异均有统计学意义(P＜0.05).两组治疗后ALT、AST、TBil均较治疗前降低,且研究组低于对照组,差异有统计学意义(P＜ 0.05).结论 拉米夫定联合干扰素α-1b治疗小儿慢性乙型肝炎可以明显改善患儿的T淋巴细胞亚群,且HBV DNA、HBsAg、HBeAg水平得到明显改善,具有显著的抗肝纤维化的作用.

目的 比较高灵敏度与普通荧光定量PCR技术在慢性乙型肝炎(CHB)患者抗病毒疗效监测中的区别.方法 对49例CHB患者141份采用普通PCR检测结果为阴性(<500 IU/mL)标本,采用高敏PCR试剂进行复查.结果 根据高敏PCR血清HBV DNA定量检测结果,141份采用普通PCR检测结果阴性标本存在以下5种状况,即>500、20～500、10～20、<10 IU/mL和阴性,比例分别为13.5％、31.9％、18.4％、30.5％和5.7％.随着血清HBV DNA水平的逐渐增高,各组ALT、AST水平和异常升高率也存在逐渐增加的趋势,但是仅仅各组AST水平差异有显著意义(P<0.05).随着血清HBV DNA水平逐渐降低,各组血清HBeAg阳性率和水平也存在逐渐降低的趋势.其中,HBV DNA阴性组血清HBeAg阳性率显著低于10～20 IU/mL组和>500 IU/mL组(P<0.05).结论 与普通PCR技术相比,高灵敏度的HBV DNA荧光定量PCR技术在CHB患者抗病毒治疗监测中准确性更高.

Objective To observe the effect of aptamer-siRNA nucleic acid compound on the apoptosis of K562 cells in human chronic myelogenous leukemia (CML)and explore its acting mechanisms.Methods K562 cells were transfected with different concentrations of aptamer-siRNA solution.The effects of aptamer-siRNA on the proliferation and apoptosis of K562 cells were detected by MTT method and AnnexinV/PI double staining method,respectively.The effects of aptamer-siRNA on the expressions of bcl-2,Bax and casepase-3 at protein and mRNA levels in K562 cells were detected by Western blot and RT-PCR method,respectively.Results Compared with the control group,the proliferation of K562 cells was significantly inhibited,early apoptosis rate of K562 cells increased significantly,the expression levels of bcl-2 protein and mRNA were significantly decreased,while the expression levels of Bax and caspase-3 protein and mRNA were significantly increased after transfection with aptamer-siRNA (P <0.05).Aptamer-siRNA nucleic acid complex at the concentration of 50 -250 μmol/L)had a significant dose-effect relationship on bcl-2,Bax and caspase-3 mRNA.Conclusion Aptamer-siRNA nucleic acid compound can promote the decreased number of bcl-2 gene and the growth of Bax and caspase-3 genes,thus promoting the apoptosis of K562 cells.

目的 观察清热抗毒煎剂联合利巴韦林治疗手足口病普通病例的临床疗效,并探讨其相关机制.方法 将120例手足口病普通患儿随机分为治疗组和对照组,每组60例.治疗组给予清热抗毒煎剂联合利巴韦林,对照组单独给予利巴韦林,观察2组的临床疗效.检测2组治疗前后白细胞(WBC)计数、C-反应蛋白(CRP)、血清免疫球蛋白IgA、IgG、IgM及补体C3、C4的含量.结果 治疗组症状消退时间短于对照组(P<0.05);治疗组临床有效率高于对照组(P<0.05);治疗后2组血清WBC计数、CRP均有降低,治疗组下调更为明显(P<0.05);治疗后2组血清IgA、IgG表达均有升高,治疗组上调更为明显(P<0.05).结论 清热抗毒煎剂联合利巴韦林治疗手足口病普通病例能够显著提高临床疗效,缩短病程;其机制可能与抑制机体炎性反应、调节机体体液免疫功能相关.

Objective To investigate the distribution characteristics of HBsAg quantification in different clinical status after chronic hepatitis B virus (HBV) infection and its correlation with HBV DNA level.Methods A cross-sectional study was performed for 268 patients with HBV infection who did not receive antiviral therapy in The Fifth Hospital of Shijiazhuang from January 2013 to December 2015.According to the stage of HBV infection,these patients were divided into immune tolerance group with 29 patients,immune clearance group with 169 patients,low-or non-replication group with 19 patients,and reactivation group with 51 patients.Roche electrochemical luminescence was used to measure serum HBsAg quantification and PCR was used to measure HBV DNA level.Logarithmic transformation was performed for the results of HBsAg quantification and HBV DNA level,and a statistical analysis was performed for the results of different stages.The Kruskal-Wallis H rank sum test was used for the analysis of multiple groups,and the Mann-Whitney U test was used for further comparison between any two groups;a Spearman rank correlation analysis was also performed.Results HBsAg quantification varied in different stages of HBV infection (x2 =71.830,P ＜ 0.001);the immune tolerance group had significantly higher HBsAg quantification than the other three groups (all P ＜ 0.05),and the immune clearance group had significantly higher HBsAg quantification than the low-replication group and the reactivation group (all P ＜ 0.05).There was a significant difference in HBV DNA level between groups (x2 =117.028,P ＜0.001);the immune tolerance group had a significantly higher HBV DNA level than the other three groups (all P ＜ 0.05),the immune clearance group had a significantly higher HBV DNA level than the low-replication group and the reactivation group (all P ＜ 0.05),and the reactivation group had a significantly higher HBV DNA level than the low-replication group (both P ＜ 0.05).HBsAg quantification was correlated with HBV DNA level in immune clearance stage (r =0.559,P ＜ 0.001) and reactivation stage (r =0.338,P =0.014).Conclusion HBsAg quantification and HBV DNA level in immune tolerance stage are higher than those in the other three stages,and HBsAg quantification is correlated with HBV DNA level in immune clearance stage and reactivation stage.

With the development and application of the double antigen sandwich method for quantification of hepatitis B core antibody (HBcAb) in recent years, there is increasing knowledge of the ability of HBcAb to reflect the body′s anti-viral capability. This article introduces the commonly used measurement methods for HBcAb and the new trends in HBcAb measurement and summarizes the association of serum HBcAb level with viral antigen and the body′s immune response, as well as research advances in effective prediction of antiviral effect with baseline HBcAb measurement before antiviral therapy. It is also pointed out that the clinical application of HBcAb needs further investigation.

目的:分析石家庄地区乙型肝炎病毒(HBV)基因型分布以及核苷(酸)类似物耐药基因突变状况.方法:对2011年1月-2013年12月在石家庄市五医院就诊的遵医嘱口服核苷(酸)类似物6个月以上的354例慢性乙型肝炎患者进行HBV P区基因序列测定.结果:HBV B型为17例(4.8％),C型为337例(95.2％);共检出突变182例,在B型中检测出有10例突变(58.8％),在C型中检测出有172例突变(51.1％),两组间突变率无显著差别;182例HBV P区存在突变的病例中以rtL180M+ rtM204I/V/S联合突变最多,占突变比例24.2％;其次是rtM204I/V/S单位点突变,占突变比例20.3％,这两种突变模式的比例达到44.5％;第3位是rtL180M+ rtM204I/V/S再加上其他一个或多个位点的突变,占突变比例12.6％.结论:石家庄地区HBV基因型以C型为主;核苷(酸)类似物耐药基因突变以拉米夫定和替比夫定耐药最常见.

Objective To detect the effects of CA 125 on cellular immune function of the ovarian cancer ( OC) patients in vitro.Methods Totally 58 patients with confirmed OC in our hospital from May 2012 to June 2013 were divided into three groups based on the CA 125 levels.Meanwhile, 20 healthy subjects (control group) and 20 patients with benign gynecological diseases ( benign diseases group ) were also included .The in vitro abil-ity of healthy human lymphocytes in secreting interferon-γ( IFN-γ) under the stimulation of phytohemagglutinin ( PHA) was observed by cell culture , enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immuno-spot (ELISPOT).Results In OC patients, the count and percentage of lymphocytes decreased gradually with elevated levels of CA 125 .The concentration of IFN-γin the supernatant of cultured lymphocytes from healthy in-dividuals decreased significantly under the stimulation of PHA and high concentration of CA 125 [(36.16 ± 16.89 ) ng/ml] compared with that of PHA alone [ (45.68 ±18.01 ) ng/ml] or PHA and low concentration of CA125 [(46.22 ±19.30)ng/ml] (both P<0.05).Compared with the control group (spots number 573.0) and the benign diseases group ( spots number 523.0 ) , the high ( spots number 108.2 ) and intermediate ( spots number 371.6 ) concentration of CA 125 had significant inhibitory effect on the cellular immune system , as detec-ted by ELISPOT ( P<0.01 , P<0.05 , respectively ) .After removal of CA125 from the serum by antibody , the inhibitory effect was weakened significantly .Conclusions The concentration of CA 125 in OC patient is nega-tively correlated with the number and activity of lymphocytes , in particular their abilities in releasing cytokines . CA125 is not only a biomarker of OC but also may serve as an immunosuppressive factor .

目的 分析石家庄地区L-核苷类抗病毒药物耐药突变情况.方法 对石家庄市第五医院就诊的慢性乙型肝炎患者119例进行乙型肝炎病毒逆转录酶区(HBV P区)基因序列测定.结果 119例患者HBV基因分型中B型5例(4.2％),C型114例(95.8％).在B型中4例rtM204I突变;C型中73例rtM204I突变,40例rtM204V突变,1例rtM204I/V突变.119例存在204位点变异的慢性乙型肝炎患者中,共有9种突变模式,其中rtM204I单位点突变占总突变比例的23.5％(28/119),rtM204V单点突变占总突变比例的6％(7/119),rtL180M+rtM204I联合突变占总突变比例的22.7％(27/119),rtL180M+ rtM204V联合突变占总突变比例的27.8％(33/119).结论 慢性乙型肝炎核苷类药物抗病毒治疗过程中,及时监测HBV P区基因耐药情况,尤其是出现rtM204I/V突变后,很容易联合其他位点突变.

目的：探讨石家庄地区重度慢性乙型肝炎患者HBV耐药突变的状况。方法对51例2012年1月至2013年12月在石家庄市五医院就诊的重度慢性乙型肝炎患者进行HBV P区基因序列测定。结果51例重度慢性乙型肝炎患者中，25例存在HBV P区发生突变，突变率为49·．0％。突变率在HBeAg阳性组和阴性组之间无显著差别（ P ＞0．05）。 HBV P区突变组血清HBV DNA水平显著高于未突变组（ P ＜0．05），ALT、AST和TBIL在突变组也显著升高（ P ＜0．05）。对25例存在HBV P区基因突变患者进行突变模式分析，居于前四位的突变模式的总和占所有突变的72％。突变模式主要为rtM204I／V单独突变以及联合rtL180M和（或）其他突变位点、rtA181V／T单独突变以及联合rtN236T或rtM250V／L突变。结论重度慢性乙型肝炎患者服用拉米夫定及阿德福韦酯等进行抗病毒治疗过程中，应实时监测HBV P区耐药突变状况，及时调整用药。

OBJECTIVETo determine the changes in the airway inflammation-related cytokine/chemokine profiles after exposure to cigarette smoke (CS) and smoking cessation (SC).METHODSA total of 18 male C57BL/6 mice were equally divided into three groups: CS group, SC group, and normal control group. The airway resistance, lung morphology, and collagen deposition around airways were determined. HE staining and Masson trichrome staining were used for histopathological analysis. The inflammatory cells in bronchoalveolar lavage fluid (BALF) were assessed. The inflammation-associated cytokines were determined using real-time PCR and immunohistochemistry. Expressions of CXCR3 ligands including the CXCL9, CXCL10, CXCL11 and other cytokines in lung tissue and BALF were also analyzed.RESULTSThe airway resistance significantly increased in both CS group and SC group when compared with the normal control group. Lung pathological scores in both CS group and SC group were also higher than that in the normal control group, while there was no significant difference between the CS group and SC group. Inflammatory cells including the neutrophils, macrophages, and lymphocytes also increased in both the CS group and SC group at both mRNA and protein levels. The mRNA levels of CXCL9, CXCL10, MMP9, and MMP12 were significantly higher in CS group and SC group than those in the normal control group (all P<0.05). The protein expression levels of CXCL9, CXCL10, CXCL11, MMP2, MMP9, MMP12, and TGF-Β1 were significantly higher in CS group and SC group than those in the normal control group (all P<0.05). Compared with the normal control group,the concentrations of CXCL9, CXCL10, CXCL11, IL-8, and TGF-Β1 in the BALF supernatants of the CS group and SC group significantly increased (P<0.05); in addition, the IL-6 and TNF-Α concentrations also increased in the CS group (both P<0.05).CONCLUSIONSCS exposure triggers inflammatory cell flux and accumulation in the lung parenchyma and BALF. As a consequence, the inflammatory cytokines increase dramatically. After CS, the cytokines/chemokines can decrease, but is still higher than in non-smokers.

目的 应用血清PG Ⅰ、G-17和PG Ⅰ/PGⅡ比值及Hp感染状况,对早期胃癌、萎缩性胃炎患者胃粘膜状况的评价,探讨胃癌高危人群的非侵袭性血清学筛查方法.方法 内镜和组织病理学确诊胃癌患者65例,内镜确诊萎缩性胃炎患者70例、正常对照组50例.ELISA法检测各组血清PG Ⅰ、PGⅡ、G-17及Hp-IgG.结果 胃体萎缩时,胃癌组血清PG Ⅰ水平及PG Ⅰ/PGⅡ比值降低,与对照组比较均具显著差异(P＜0.01),与胃炎组比较亦均有显著差异(P＜0.05).多灶性萎缩性时,胃癌组血清PG Ⅰ水平和PG Ⅰ/PGⅡ比值显著降低,与正常对照组比较具非常显著差异(P＜0.01和P＜0.001),与胃炎组比较亦具有显著差异(P＜0.05和P＜0.01),血清G-17水平显著降低,与正常对照组和胃炎组比较具有统计意义(P＜0.001和P＜0.05),且多灶性萎缩病变在胃癌中的比例明显多于胃炎组.而Hp感染不影响胃癌患者血清PG Ⅰ和G17的水平.结论 低水平血清PG Ⅰ、PG Ⅰ/PGⅡ比值和G-17提示患者可能有胃癌高风险的多灶性萎缩病变,血清PGs和G-17的检测可作为胃粘膜萎缩的非侵袭性检查方法.

目的 探讨循环DNA在急性脑梗死患者中的应用价值.方法 采集20例急性脑梗死患者(研究组)发病后24 h内静脉血2 ml,采用双重荧光定量PCR技术检测血浆DNA水平,同时检测血清超敏C-反应蛋白(hs-CRP).采集同期20例健康体检者静脉血为对照组.结果 治疗前研究组血浆循环DNA水平显著高于对照组(t=5.237,P=0.000),研究组hs-CRP显著高于对照组(t=4.890,P=0.000);治疗后循环DNA显著下降(t=4.461,P=0.000);hs-CRP治疗前后差异有统计学意义(t=3.964,P=0.000).脑梗死患者血浆循环DNA水平与其血清hs-CRP水平呈正相关(r=0.613,P=0.022).结论 循环DNA可作为反映脑梗死后脑组织细胞出现损伤及严重程度的一项指标.

目的 探讨高血压性脑出血患者血浆循环DNA与纤维蛋白原的相关性.方法 采集30例高血压性脑出血患者发病后24h静脉血,采用双重荧光定量PCR技术检测血浆循环DNA水平,并同时检测血浆纤维蛋白原水平.采集30例健康体检者静脉血作为对照组.结果 脑出血患者血浆循环DNA水平(313.2±224.7)mg/L,显著高于对照组的(26.1±5.9)mg/L,差异有统计学意义(P＞0.05).脑出血患者轻、中、重度3组血浆循环DNA水平之间差异有统计学意义(P＜0.05),重度组血浆循环DNA水平显著高于轻度组和中度组(P＜0.05),中度组显著高于轻度组(P＜0.05).脑出血患者血浆纤维蛋白原水平(4.25±1.51)g/L,显著高于对照组的(2.76±0.85)g/L,差异有统计学意义(P＜0.05);重度组血浆纤维蛋白原水平显著高于轻度组和中度组(P＜0.05);中度组与轻度组间血浆纤维蛋白原水平差异无统计学意义.脑出血患者血浆循环DNA水平与其纤维蛋白原水平呈显著正相关(r=0.632,P＜0.05).结论 血浆循环DNA可作为反映高血压性脑出血后脑组织细胞损伤及严重程度的灵敏指标.

Objective To explore the related factors of circulating DNA level in patients with chronic HBV infection. Methods One hundred and forty-five patients with chronic HBV infection were enrolled in the study, including 30 cases with mild chronic hepatitis B, 30 cases with moderate chronic hepatitis B, 25 cases with severe chronic hepatitis B, 30 cases with HBV-induced liver cirrhosis, 30 cases with hepatocarcinoma. Thirty healthy persons were also enrolled as control. The level of plasma circular DNA was detected by duplex real-time polymerase chain reaction assay. Results As the condition of patients with chronic hepatitis B exacerbated, the level of circulating DNA increased gradually,but it reduced in patients with HBV-induced liver cirrhosis, then increased significantly in patients with hepatocarcinoma. There were significant differences among groups(P<0.05). The level of circulating DNA in plasma HBeAg positive group(68.79±14.23 ng/mL) was significantly higher than that of HBeAg negative group(23.68±12.56 ng/mL)(t=2.162, P=0.032).There was a significantly positive relationship between circulating DNA and alanine aminotransferase and aspartate aminotransferase (r=0.612;0.632,P=0.014;0.021). There was no relationship between circulating DNA and HBV DNA in serum(P> 0.05). Conclusion The level of circulating DNA in patients with chronic HBV infection had no relationship with viral replication, but reflects the degree of hepatocyte inflammatory necrosis.

血浆循环DNA是一种游离的细胞外DNA，主要存在于血液、滑膜液等体液中，生理情况下体内循环DNA的含量很少，当机体处于炎性反应、组织细胞损伤或肿瘤时，循环DNA水平会相应的增加［1］，增加的循环DNA主要来自破坏的细胞释放至外周血或者肿瘤细胞主动分泌到外周循环，因而定量检测循环DNA含量的变化有助于疾病的诊断及病情的监测。艾滋病是由HIV感染引起的慢性传染病。 HIV主要侵犯、破坏人体CD4＋T淋巴细胞，导致机体免疫功能受损乃至缺陷，最终并发各种严重机会性感染和肿瘤。因此，早期监测HIV感染者病情变化对于并发症的发生、提高生存质量、延长患者寿命有着重要意义。本研究采用荧光定量PCR技术检测各期HIV感染者外周血循环DNA的含量，探讨循环DNA水平作为HIV感染者病情判断的指标。

Objective To study the effect of Ruangan Huajian granule on the expressions of apoptosis related gene Bcl-2 and Bax in liver of C57 mice with experimental hepatic fibrosis. Methods Forty C57 mice were randomly divided in to four groups : normal control group, model control group, low-dose of Ruangan Huajian granule group and high-dose of Ru angan Huajian granule group. There were 10 mice in every group. C57 mice with experimental hepatic fibrosis were induced by intraperitoneal injection of 10% CCl 4 . The expressions of Bcl-2 and Bax in liver tissues of mice were observed by immune histochemical technique. The score of fibrosis stages in the liver tissue was also analyzed. Results The expressions of Bcl-2 and Bax were mainly in the eosinophilic liver cells, especially in model control group. The eosinophilic liver cells were significantly reduced in low-dose and high-dose of Ruangan Huajian granule groups. The score of fibrosis stages and the positive expressions of Bcl-2 and Bax were significantly higher in model control group than those of normal control group ( P 0.05), but the value of Bcl-2/Bax was significantly lower in model control group than that of normal control group ( P 0.05). The positive expression of Bcl-2 was significantly higher in low-dose and high-dose of Ruangan Huajian granule groups than that of model control group and normal control group ( P 0.05). The positive expression of Bax was significantly lower in low-dose and high-dose of Ruangan Huajian granule groups than model control group, but it significantly enhanced than that of normal control group ( P 0.05). The value of Bcl-2/Bax was significantly higher than that of model control group and normal control group ( P 0.01). Conclusion Ruangan Huajian granule can modulate the expressions of Bcl-2 and Bax in liver tissues, and inhibit the liver cell apoptisis, showing an anti-fibrotic effect.

目的对阿德福韦酯治疗过程中发生病毒学突破的慢性乙型肝炎患者进行rtN236位点、rtA181位点以及两者联合突变的分析,观察各自的病毒学特征是否有差异。方法选择42例慢性乙型肝炎患者,收集病毒学突破后的血清标本,采用罗氏cobase601电化学发光分析仪检测血清HBsAg含量;采用ABI7300实时荧光定量PCR仪进行血清HBVDNA定量检测;采用美国雅培C8000全自动生化分析仪检测血清ALT。结果 42例患者中发生rtA181位点突变27例,rtN236位点突变6例,两者联合突变9例;单纯rtA181变异组血清表面抗原明显低于单纯rtN236变异组和rtA181+rtN236变异组(P=0.04);rtA181+rtN236变异组血清HBVDNA和ALT明显高于单纯rtA181变异组和单纯rtN236变异组(P=0.03,0.04);所有患者乙型肝炎病毒基因型中只有3例是B型,而且这3例均在单纯N236变异组,其他均为C型。结论阿德福韦耐药rtN236T突变株与rtA181T突变株有各自的临床特点,在平时的检测和诊断中应引起足够重视。