目的 探究白细胞介素-28B(IL-28B)基因多态性与慢性乙型肝炎病毒(HBV)感染相关肝硬化发生及预后的关联.方法 选取2021年10月-2022年12月于杭州市西溪医院就诊的慢性HBV感染相关肝硬化患者120例为肝硬化组,另招募同期性别、年龄匹配的体检健康人群120名为对照组,其中肝硬化患者根据疾病进展情况分为代偿期组和失代偿期组;检测血清IL-28B水平和肝功能标志物,并采用聚合酶链反应(PCR)法和直接测序法检测外周血IL-28B基因rs12979860、rs8099917位点多态性.结果 肝硬化组血清IL-28B水平低于对照组,肝功能指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和总胆红素(TBIL)均高于对照组(P＜0.05);肝硬化组和对照组rs12979860位点、rs8099917位点基因型和等位基因差异有统计学意义(P＜0.05),其中rs12979860位点C等位基因、rs8099917位点T等位基因是HBV感染相关肝硬化的影响因素(P＜0.05);失代偿期组患者血清IL-28B低于代偿期组,ALT、AST及TBIL均高于代偿期组(P＜0.05).结论 IL-28B基因rs12979860、rs8099917位点多态性与HBV感染相关肝硬化发生和预后有关,可能通过影响血清IL-28B水平参与慢性HBV感染相关肝硬化发生发展.

目的 探讨双重血浆分子吸附系统(double plasma molecular absorb system,DPMAS)联合甲泼尼龙琥珀酸钠(methylprednisolone sodium succinate,MPSS)治疗慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)的疗效.方法 选取 2020 年 1 月至 2022 年 1 月于杭州市西溪医院诊治的ACLF患者 102 例,采用简单随机法将其分为观察组和对照组,每组各 51 例.对照组患者给予MPSS静脉滴注治疗,观察组患者在对照组基础上联合DPMAS治疗.比较两组患者的临床疗效、肝功能[天冬氨酸转氨酶(aspartate aminotransferase,AST)、丙氨酸转氨酶(alanine aminotransferase,ALT)、总胆红素(total bilirubin,TBIL)]、凝血功能[凝血酶原时间(prothrombin time,PT)、活化部分凝血活酶时间(activated partial thromboplastin time,APTT)、纤维蛋白原(fibrinogen,FIB)]和甲状腺激素[游离三碘甲腺原氨酸(free triiodothyronine,FT3)、游离甲状腺素(free thyroxine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)].结果 观察组患者的总有效率显著高于对照组(χ2=4.194,P=0.041).治疗后,两组患者的AST、ALT和TBIL均显著低于本组治疗前,PT、APTT均显著短于本组治疗前,FIB、TSH均显著高于本组治疗前(P<0.05),观察组患者的AST、ALT和TBIL均显著低于对照组,PT、APTT 均显著短于对照组,FIB、TSH 均显著高于对照组(P<0.05).结论 DPMAS 联合 MPSS 治疗ACLF疗效显著,可改善患者的肝功能、凝血功能和甲状腺激素水平.

目的 研讨心理干预对妊娠合并慢性乙型肝炎患者心理状态和妊娠结局的影响.方法 选取2018年6月—2019年11月妊娠合并慢性乙型肝炎患者120例,采用随机数字表法分为心理干预组和对照组各60例.心理干预组予以综合性心理干预,对照组予以常规干预措施,记录受试者干预前、后的焦虑(SAS)与抑郁(SDS)分值、妊娠结局及分娩并发症,并进行统计分析.结果 心理干预组在干预后SAS与SDS分值比对照组显著降低,差异有统计学意义(P＜0.05).心理干预组剖宫产率及早产、新生儿窒息、羊水粪染发生率显著低于对照组,差异有统计学意义(P＜0.05).结论 在孕期实施心理干预有助于改善合并慢性乙型肝炎孕妇的不良心理状态,并降低不良妊娠结局及分娩并发症的发生率.

目的 探讨慢性乙型病毒性肝炎患者发生慢加急性肝衰竭的危险因素.方法 选取2018年1月至2020年6月在杭州市西溪医院诊断治疗的慢性乙型肝炎患者130例的临床资料进行回顾性分析,130例患者中35例发生慢加急性肝衰竭,为肝衰组,95例为非肝衰组.记录患者的一般资料及临床资料,单因素及多因素分析慢性乙型病毒性肝炎患者发生慢加急性肝衰竭的危险因素.结果 单因素分析结果显示,HBV DNA≥107拷贝/L,有家族史,合并感染,肝功能分级C级的患者发生肝衰竭的发生率更高,差异有统计学意义(P<0.001);肝衰竭组TBiL更高,清蛋白水平更低,国际标准化比率更高,凝血酶原活动度更低,与非肝衰组比较,差异有统计学意义(P<0.001).多因素分析结果显示,HBV DNA≥107拷贝/L,感染,肝功能分级C级,TBiL≥171μmol/L,国际标准化比率≥1.5是慢性乙型病毒性肝炎发生慢加急性肝衰竭的独立危险因素,PTA>40％,清蛋白>20 g/L是保护因素(P<0.001).结论 慢加急性肝衰竭是慢性乙型病毒性肝炎严重的并发症,患者预后较差.影响慢性乙型病毒性肝炎患者发生慢加急性肝衰竭的因素较多,对于具有高危因素的患者,应积极预防慢加急性肝衰竭的发生.

目的 探讨外周血微小RNA(miR)-146基因多态性与乙型肝炎病毒感染患者肝硬化的易感性及其水平与病情严重程度的关系.方法 选择杭州市西溪医院肝病科2018年5月-2020年6月收治的乙型肝炎肝硬化患者100例纳入研究组,选择同期医院收治的乙型肝炎非肝硬化患者60例作为对照组.采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测两组miR-146a基因rs2910164位点基因型及表达水平,比较不同基因型患者乙型肝炎病毒载量、血清谷丙转氨酶(ALT)、谷草转氨酶(AST)及总胆固醇(TC)水平.结果 研究组miR-146a基因rs2910164位点CC基因型、C等位基因频率均高于对照组(P＜0.05);100例慢性乙型肝炎肝硬化患者根据病情程度分为Child-pugh(A、B、C)三组,Child-pugh C组CC基因型频率高于Child-pugh A组及Child-pugh B组(x2=18.291、11.431,P均＜0.001);miR-146a基因rs2910164位点CC基因型患者miR-146 mRNA、AST、ALT、TC水平高于TT/TC患者(P＜0.05),HBV-DNA水平比较差异无统计学意义.结论 miR-146a基因rs2910164位点多态性与HBV感染者发生肝硬化风险相关,miR-146 mRNA水平对肝硬化程度判断具有一定的价值.

ObjectiveTo investigate the efficacy of Peg-interferon alphaα sequential combinated with Nucleos(t)ide analogues on HBeAg positive chronic hepatitis B patients with high serum hepatitis B viral loads compared with Peg-interferon alphaα alone under response-guided therapy. Methods47 CHB patients with HBeAg-positive and HBVDNA>1×107 IU/ ml used Peg-interferon alphaα antiretroviral therapy after 12 weeks, then divided them into joint group and monotherapy group according to HBV DNA decline is greater than 2 lg IU / ml or not:joint group received Peg-interferon alphaα combinated with one nucleoside drugs,monotherapy group continued to receive Peg-interferon alphaα alone,both groups were treated to 48 weeks. Observed boths groups of ALT normalization,HBV DNA negative rate,serum HBeAg and HBsAg negative/seroconversion rate after the treatment of 24weeks and 48 weeks.ResultsHBV DNA undetectable rate of 24weeks and 48weeks in joint group were higher than that of single treatment with Peg-IFN group(56.25% & 41.94%、93.75% & 70.97%),however the differences between them had no statistical significance (χ2=0.8685、3.2702,P>0.05); At 24 weeks and 48 weeks,the HBeAg and HBsAg quantitative decline extent of joint group were faster than that of monotherapy group.Conclusions Response can be improved under the strategy of response-guided therapy on high serum hepatitis B viral loads and HBeAg positive chronic hepatitis B patients who have poor response treat with Peg-interferon alphaα sequential combinated with Nucleos(t)ide analogues,but long-term effect remains to be further studied.

Objective To evaluate the clinical significance of plasma procalcitonin (PCT) and C-reactive protein(CRP) in patients with malaria. Methods A total of 33 cases of malaria were selected and divided into the severe malaria group and non-severe malaria group. The PCT, CRP, Hb, ALT, TB, PT and serum potassium between two groups were detected and compared. Results The PCT of the severe malaria group was significantly higher than non-severe malaria group as well as the CRP (t=-3.75,-4.68, P<0.05). The concentration of Hb of severe malaria group was significantly lower than non-severe malaria group (t=2.36,P<0.05). There was no statistical difference of TB, ALT, PT and serum potassium between two groups (t=-1.55,-0.16,-0.94,-0.52, P>0.05). Conclusion PCT and CRP can be as indexes to assess the severity of malaria.

Objective To compare the 144-week efficacy of de-novo combination therapy with lamivudine (LAM) and adefovir dipivoxil (ADV) to that of optimize combination for chronic hepatitis B (CHB) patients.Methods A total of 83 cases with CHB were divided into 3 groups:the de-novo combination group (group A,28 cases),after 24-week mono-therapy to optimize combination group (group B,32 cases),after 48-week mono-therapy to optimize combination group(group C,23 cases).The virological,serological,biochemical indicators at baseline,week 24,48,96 and 144 were detected respectively.The gene resistance mutations of of HBV P region were analyzed for the patients whose HBV DNA were still positive (HBV DNA≥ 103 copies/mL) at week 96,144,respectively.Data of 3 groups were analyzed by the pharmacoeconomics cost-effectiveness analysis.Results HBV DNA negative conversion rates in 3 groups after 96-week therapy were 92.9％,90.6％,87.0％.There were no significant differences among 3 groups (x2 =0.509,P ＞0.05); HBV DNA negative conversion rates in 3 groups after 144-week therapy were 96.4％,93.8％,91.3％,and the differences among them were not statistically significant(x2 =0.590,P ＞ 0.05).There was 1 patient with virologic breakthrough at 144-week treatment in group A.The mutation sites in HBV P genes were L180M and M204V.HBeAg-positive patients with seroconversion rates in 3 groups after 96-week therapy were 27.8 ％,19.2％,35.7 ％,and the differences were not statistically significant (x2 =1.340,P ＞ 0.05).The seroconversion rates in 3 groups at week 144 were 38.9％,30.8％,42.9％,and the differences were not statistically significant (x2 ＝ 0.364,P ＞ 0.05).The ALT normalization rates in 3 groups at week 96 were 94.1％,90.9％,88.9％,and the differences were not statistically significant (x2 =0.303,P ＞ 0.05) ; The ALT normalization rates in 3 groups after 144-week therapy were 100.0 ％,95.5 ％,94.4％,and the differences were not statistically significant(x2=0.911,P ＞ 0.05).The cost-effect analysis showed that the initial combination group was superior to the optimize combination group.Conclusions The effects of virology negative conversion rate,seroconversion rate,ALT normalization rate and reduction the incidence of drug resistance in LAM + ADV de-novo combination therapy and optimized combination therapy are good,but optimized combination therapy is more economic than de-novo combination therapy,so optimized combination therapy can be recommended for use.

Though nucleos(t)ide analogs (NAs) and interferon have been considered as the main drugs for treatment of chronic hepatitis B,the monotherapy of NAs or interferon is prone to induce low or non-response,antiviral drugresistance and unbearable adverse reactions.Therefore antiviral drugs combination treatment for chronic hepatitis B re-at-tracts clinical attention.This paper reviews the application of antiviral drugs combination treatment for chronic hepatitis B.

[Objective]To explore the effect of bilirubin on the cell proliferation and cell cycle in hepatic stellate cell(HSC)-T6. [Methods]The immortalized rat hepatic stellate cell line HSC-T6 was commonly cultured after revival,and was allocated into normal control group and treatment groups with different concentrations bilirubin.The cell proliferation was assessed by using MTT assay,and cell cycle was analyzed by flow cytometry. [Results]①After intervention of different concentration bilirubin(the extreme concentration was 10 μmol/L,30 μmol/L,50 μmol/L,70 μmol/L,100 μmol/L respectively),the A value increased significantly(compared with the control group,all P<0.05)and showed a dose-dependent manner.②In cell cycle progression,the cell at G_0/G_1 phase decreased obviously but increased at S and G_2/M phase when bilirubin concentration at 10 μmol/L,50 μmol/L,100 μmol/L(compared with the control group,P<0.05). [Conclusion]Bilirubin can promote the proliferation of HSC-T6.

Objective To evaluate the influence to detect ascites bacteria with 16S rRNA PCR-microarray in patients with spontaneous bacterial peritonitis( SBP) after treated with antibiotics.Methods Compared to ascites bacterial culture,ascites bacterial 16S rRNA genes were detected by PCR-microarray in 76 cases of suspected SBP with chronic liver diseases,and the difference of ascites bacteria positive rates between two groups with and without antibiotics were analyzed.Results Of the 76 ascites samples in SBP patients,there were 17(22.37％ ) ascites bacteria positive detected by PCR-microarray,and 6(7.89％ ) ascites bacteria positive detected by bacterial culture.The positive rates of former were much higher(x2 =18.05,P ＜0.01).In groups with and without antibiotics,the positive rates of bacterial detection with PCR-microarray were 19.35％ and 24.44％ respectirely,there was no statistical difference between the two groups( x2 =0.274,P ＞ 0.05) ; and the positive rates of bacterial detection with bacterial culture were 0(0/31 ) and 13.33％(6/45) respectively,there was statistical difference between the two groups(x2 =4.488,P ＜ 0.05).Conclusions Compared to the ascites bacterial culture,less influence is found in ascites bacteria detection with 16S rRNA PCR-microarray in patients with SBP after treatment with antibiotics.

Objective To compare the efficacy of combination therapy with lamivudine(LAM) and adefovir dipivoxil(ADV) for LAM-poorly responsed and LAM-resistant to those de novo combination therapy for chronic hepatitis B (CHB).Methods Twenty CHB patients with poorly response to LAM were selected as group A,20 LAM-resistant patients as group B,other 20 CHB patients received combination therapy for the first time as group C.All the patients were given LAM ( 100 mg/d) and ADV (10 mg/d) for 48 weeks,the level of HBV DNA,ALT and HBeAg in patients were detected and HBV mutation was detected for whose HBV DNA were still positive after treatment.Results The negative rate of HBV DNA in group A,B and C were 85％,80％,90％ ( F =0.784,P ＞ 0.05),and the negative rate of HBeAg were 31.3 ％,30.8 ％,33.3 ％ ( F =0.025,P ＞ 0.05) in group A,B,C,respectively.The level of HBV DNA and ALT decreased remarkably after 48 combination treatment in group A,B,C ( t =9.706,10.871,11.807,P ＜ 0.01 ;t =2.157,2.109,2.653,P ＜ 0.05).HBV DNA in 9 patients were still positive after treatment,but no HBV mutations were detected in three groups.Concusions The combination therapy of LAM and ADV can effectively inhibit HBV replication,improve ALT level in CHB patients,but the efficacy in HBeAg seroconversion rate are not obvious.The efficacy of combination therapy in three groups are similar.No HBV mutations are detected for three groups.

Antiviral therapy is regarded as the key of chronic hepatitis B treatment. In terms of virology, biochemics and histology, no matter nucleoside/nucleotide analogues are used in monotherapy or combination therapy, these medicines can benefit all the patients, whose HBeAg are positive or negative. However, the drug-resistance occurs frequently in the long-term application, limiting its clinic effects. This thesis aims to give general introduction of the current status of chronic hepatitis B treatment with nucleoside/nucleotide analogues by monotherapy and combination therapy.

[目的]观察恩替卡韦联合复方鳖甲软肝片治疗慢性乙型肝炎肝纤维化临床疗效。[方法]将96例慢性乙型肝炎肝纤维化患者随机分为恩替卡韦片与复方鳖甲软肝片联合治疗(治疗)组及恩替卡韦片(对照)组,每月查肝功能等生化指标,每3个月检测HBVM、HBVDNA,分别于治疗前和治疗12个月后放射免疫法各检测1次血清肝纤维化指标。[结果]两组患者治疗前后肝功能好转、HBVDNA水平下降,但两组治疗后比较无明显差异(P>0.05),治疗组肝纤维化程度改善更明显(P<0.05)。[结论]恩替卡韦片与复方鳖甲软肝片联合治疗在抗纤维化程度方面优于单用恩替卡韦片。

Objective To evaluate clinical applications on the quantitative PCR detection of ascitic bacterial 16S rRNA genes in the diagnosis of spontaneous bacterial peritonitis with non-neutrophil ascites. Methods The bacterial 16S rRNA was detected by quantitative fluorescent PCR in 64 cases of suspected spontaneous bacterial peritonitis with nonneutrophil ascites, 6 cases of chronic liver disease accompanied with non-infected ascites at the same time period, as compared to the ascitic bacterial culture. Results Bacterial 16S rRNA quantitative PCR can detect DNA duplicate as low as 10 copies. The rate of detectable bacterial 16S rRNA by quantitative PCR( 15.63% ) was significantly higher than the rate of positive bacterial culture (3.13% ) ( x2 = 5.52, P ＜ 0.05 ); and 6 cases of non-infected ascites were negative analyzed by quantitative fluorescent PCR and bacterial cultures. Conclusions The ascitic bacterial 16S rRNA quantitative PCR detection is high specificity and sensitivity for the diagnosis of spontaneous bacterial peritonitis with non-neutrophils ascites, whtich is of important clinical significance.

OBJECTIVE:To evaluate the value of ascitic bacterial 16S rRNA gene determination in the rapid diagnosis of spontaneous bacterial peritonitis (SBP).METHODS:16S rRNA gene from bacterial DNA in ascites was determined by quantitative fluorescent polymerase chain reaction (PCR) in 76 patients with suspected SBP and 6 patients with non-infectious ascites. The results were compared with those obtained from bacterial culture.RESULTS:The positive rate of SBP was 22.4% among patients detected with ascitic bacterial 16S rRNA gene determination-based quantitative fluorescent PCR, which was significantly higher than that (7.9%) in patients only received bacterial culture (P<0.05). In addition,in 6 patients with non-infectious ascites,both the 16S rRNA gene determination-based quantitative fluorescent PCR and bacterial culture showed negative results.CONCLUSIONS:16S rRNA gene determination-based quantitative fluorescent PCR can be an effective tool for the rapid diagnosis of SBP. It is more sensitive than the bacterial culture.