Current status of hepatitis C virus infection and progress in its elimination in China / 临床肝胆病杂志

Hepatitis C virus （HCV） can develop into liver cirrhosis or hepatocellular carcinoma， imposing a heavy burden on the patient’s family and the society. Hepatitis C is one of the major public threats for humans， and eliminating hepatitis C is a common goal of all humans. Direct-acting antiviral agents （DAAs） are currently a relatively safe treatment regimen for hepatitis C that can reach a relatively high cure rate and can target different HCV genotypes， making it possible to eliminate HCV infection. China actively promotes the clinical application of DAAs， accelerates drug approval， improves the accessibility of DAAs， and strengthens population intervention. National Medical Insurance Administration has gradually included DAAs in the national medical insurance directory， providing strong support for eliminating HCV infection. In response to the WHO’s goal of eliminating viral hepatitis as a public health hazard by 2030， China has successively released national strategic plans and action plans in recent years， making significant achievements in HCV infection elimination， forming a joint prevention and control system across multiple sectors of the society， and ultimately achieving the goal of eliminating HCV infection. With a focus on the current status of HCV infection in China and prominent prevention and control strategies， this article analyzes and summarizes the practical process of the prevention， control， and micro-elimination of HCV infection， in order to provide a policy reference for carrying out HCV elimination in China and help to achieve the goal of comprehensive elimination of HCV infection.

Comparison of bacterial diversity in Bactrocera cucurbitae (Coquillett) ovaries and eggs based on 16S rRNA sequencing.

Next-generation sequencing allows for fine-scale studies of microbial communities. Herein, 16S ribosomal RNA high-throughput sequencing was used to identify, classify, and predict the functions of the bacterial communities in the eggs and ovaries of Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), which is a pest that infests a variety of cucurbit fruits at different developmental stages. Taxonomic analyses indicate that bacteria associated with B. cucurbitae represent 19 phyla, which were spread across different developmental stages. Specifically, the egg microbiota had a higher alpha diversity than those of microbiota in the primary and mature ovaries. Significant differences were not observed between the primary and mature ovaries in terms of their microbiota's alpha diversities. Pseudomonadota, Deinococcota, Bacteroidota, Bacillota, and Actinomycetota were the dominant phyla in all three developmental stages of B. cucurbitae, and Pseudomonadaceae and Enterobacteriaceae were the most abundant families. Owing to the unique physiological environment of the ovaries, the diversity of their bacterial community was significantly lower than that in the eggs. This study provides new insights into the structure and abundance of the microbiota in B. cucurbitae at different developmental stages and contributes to forming management strategies for this pest.

Key points for viral hepatitis management in "Global health sector strategies on, respectively, HIV, viral hepatitis and sexually transmitted infections for the period 2022— 2030":interpretation and clinical practice / 中华检验医学杂志

In June 2022, the World Health Organization released the"Global health sector strategies on, respectively, HIV, viral hepatitis and sexually transmitted infections for the period 2022—2030". It sets more specific and large-scale goals for eliminating the major infectious diseases, which are still threatening human health. In combination with our clinical practice, present document highlights the advances and challenges in the process of implementing the suggested strategies and ways to achieve the stated goal of eliminating the viral hepatitis in China.

Studies on the level of neutralizing antibodies produced by inactivated COVID-19 vaccines in the real world

BackgroundAlthough effective vaccines have been developed against COVID-19, the level of neutralizing antibodies (Nabs) induced after vaccination in the real world is still unknown. To evaluate the level and persistence of NAbs induced by two inactivated COVID-19 vaccines in China. Methods and findingsSerum samples were collected from 1,335 people aged 18 and over who were vaccinated with COVID-19 inactivated vaccine in Peking University Peoples Hospital from January 19 to June 23, 2021, for detection of COVID-19 antibodies. The WHO standard of SARS-CoV-2 NAbs was detected. The coefficients of variation between the detection results and the true values of the NAbs detected by the WHO standard were all lower than the WHO international standard 3% after the dilution of the original and the dilution of the theoretical concentrations of 500 IU/mL, 250 IU/mL, 125 IU/mL, 72.5 IU/mL, 36.25 IU/mL and 18.125 IU/mL. On day 11-70, the positive rate of NAbs against COVID-19 was 82% to 100%; From day 71 to 332, the positive rate of NAbs decreased to 27%. The level of NAbs was significantly higher at 3-8 Weeks than at 0-3 Weeks. There was a high linear correlation between NAbs and IgG antibodies in 1335 vaccinated patients. NAbs levels were decreased in 31 of 38 people (81.6%) at two time points after the second dose of vaccine. There was no significant difference in age between the group with increased and decreased neutralizing antibody levels ({chi}2 =-0.034, P>0.05). The positive rate of NAbs in the two-dose vaccine group (77.3%) was significantly higher than that in the one-dose group (18.1%), with statistical difference ({chi}2=312.590, P<0.001). A total of 206 people who were 11-70 days after receiving the second dose were tested and divided into three groups: 18-40 years old, 41-60 years old and >60 years old. The positive rates of NAbs in three groups (18-40 years old, 41-60 years old and >60 years old) were 95.14%, 78.43% and 81.8%, respectively. The positive rate of NAbs was significantly higher in 18-40 years old than in 41-60 years old ({chi}2=12.547, P <0.01). The titer of NAbs in 18-40 years old group was significantly higher than that in 41-60 years old group (t=-0.222, P <0.01). The positive rate of NAbs in male group (89.32%) was lower than in female (91.26%), but there was no significant difference ({chi}2=0.222, P >0.05). ConclusionsThe positive rate of NAbs was the highest from 10 to 70 days after the second dose of vaccine, and the positive rate gradually decreased as time went by. There was a high linear correlation between COVID-19 NAbs and IgM/IgG antibodies in vaccinators, suggesting that in cases where NAbs cannot be detected, IgM/IgG antibodies can be detected instead. The level of NAbs produced after vaccination was affected by age, but not by gender. The highest levels of NAbs were produced between shots 21 to 56 days apart, suggesting that 21 to 56 days between shots is suitable for vaccination. Author summaryO_ST_ABSWhy was this study done?C_ST_ABSO_LIAt present, the inactivated vaccines that have been approved to market in China have passed clinical trials to prove their effectiveness and safety. But the level of neutralizing antibodies induced by vaccination in the real world remains unclear. C_LIO_LISerological testing for neutralizing antibodies against COVID-19 is important for assessing vaccine and treatment responses and comparing multiple drug candidates. We assessed the levels of neutralizing antibodies produced in populations receiving inactivated vaccines and assessed the persistence of these vaccines in producing COVID-19 neutralizing antibodies in healthy adults. C_LI What did the researchers do and find?O_LIWe collected serum samples from 1,335 people aged 18 and above who had received COVID-19 vaccine in Peking University Peoples Hospital, and divided them into two groups according to one dose of inactivated vaccine and two doses of inactivated vaccine. C_LIO_LIOur study found that the positive rate of NAbs was 66.2% in adults who received one or two doses of inactivated vaccine and 77.3% in adults who received two doses of inactivated vaccine in the real world. C_LIO_LIFrom 11 to 70 days after the second dose of vaccine, the positive rate of neutralizing antibodies against COVID-19 was 82-100%; On days 71-332, the positive rate of neutralizing antibodies decreased to 27%. C_LIO_LIThe titer and the positive rate of NAbs in 18-40 years old group were significantly higher than that in 41-60 years old group. C_LI What do these findings mean?O_LIWhat is novel is we observed that in the real world, the positive rate of neutralization antibody was the highest at 10 to 70 days after the second vaccination, and with the extension of the vaccination time, the positive rate of antibody gradually decreased. Therefore, we recommend that the third dose of vaccine be administered at day 61 to day 70 for COVID-19 neutralizing antibodies levels. C_LIO_LIWe observed that there was a high linear correlation between COVID-19 neutralization antibodies and COVID-19 IgM/IgG antibodies in vaccinators, suggesting that in cases where NAbs cannot be detected, COVID-19 IgM/IgG antibodies can be detected instead. C_LIO_LIIn our manuscript, we found that the titer and positive rate of neutralizing antibodies in 18-40 years old group were higher than those in 41-60 years old group. The level of neutralizing antibodies produced after vaccination was affected by age, but not by gender. C_LIO_LIWe also observed that the highest levels of NAbs were produced between shots 21 to 35 days apart, suggesting that 21 to 35 days between shots is suitable for vaccination. C_LI

Current status of the research on low-level viremia in chronic hepatitis B patients receiving nucleos(t)ide analogues / 临床肝胆病杂志

Nucleos(t)ide analogues (NAs), which are widely used as the first-line anti-hepatitis B virus (HBV) drugs in clinical practice, can effectively inhibit the replication of HBV DNA, significantly slow down disease progression in chronic hepatitis B (CHB) patients, and reduce the development of end-stage liver diseases such as liver failure and liver cancer. However, for some CHB patients receiving first-line NAs for 48 weeks or longer, serum HBV DNA is still persistently or intermittently higher than the lower detection of limit of sensitive nucleic acid detection reagents. After discussion by the authors, low-level viremia (LLV) is defined as follows: persistent LLV refers to the condition in which CHB patients, who receive entecavir, tenofovir disoproxil fumarate, or tenofovir alafenamide fumarate for ≥48 weeks, test positive for HBV DNA by two consecutive detections with sensitive quantitative PCR, with an interval of 3-6 months, but have an HBV DNA level of ＜2000 IU/ml; intermittent LLV refers to the condition in which patients test positive for HBV DNA intermittently by at least three consecutive detections with sensitive quantitative PCR, with an interval of 3-6 months, but have an HBV DNA level of ＜2000 IU/ml. For the diagnosis of LLV, the issues of poor compliance and drug-resistant mutations should be excluded. LLV might be associated with the increased risk of progression to liver fibrosis or hepatocellular carcinoma in patients with liver cirrhosis under NA treatment, but there are still controversies over whether the original treatment regimen with NAs should be changed after the onset of LLV. This article summarizes the incidence rate of LLV under NA treatment and the influence of LLV on prognosis and analyzes the possible mechanisms of the osnet of LLV, so as to provide a reference for the management of LLV in patients treated with NAs.

FOXP3 gene polymorphisms and hepatocellular carcinoma / 中华普通外科杂志

Objective To investigate the correlation between single nucleotide polymorphisms (SNPs) of FOXP3 gene and the susceptibility of hepatocellular carcinoma (HCC). Methods Two SNPs rs2280883 and rs3761549 of FOXP3 gene in 392 HCC patients and 372 healthy controls were analyzed by Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry (MALDI-TOF).Results At rs3761549,C allele frequency was significantly higher ( OR =1.32,95％ CI 1.03 -1.70,P =0.027) in HCC patients than healthy controls.Compared with healthy controls,HCC patients had higher frequencies of TT genotype (79.6％ ) at rs2280883 or CC genotype (77.6％) at rs3761549 of FOXP3 gene.Patients carrying rs2280883 TT genotype ( OR =1.53,95％ CI 1.10 - 2.14,P ＜ 0.00001 ) or rs3761549 CC genotype ( OR =1.92,95％ CI 1.39 - 2.64,P ＜ 0.00001 ) were more susceptible to HCC.Stratified analysis showed that rs3761549 CC genotype was significantly associated with higher incidence of portal vein tumor thrombus ( x2 =5.578,P =0.018 ),and rs3761549 TT/CT genotype was significantly associated with higher rate of tumor recurrence in HCC patients (x2 =6.561,P =0.010).Conclusions FOXP3 gene polymorphisms at rs2280883 and rs3761549 might be associated with increased susceptibility to HCC. rs3761549,CC genotype and TT/CT genotype were respectively associated with higher incidence of portal vein tumor thrombus and tumor recurrence in HCC patients.

The effect of endothelial progenitor cell transplantation on liver cirrhosis in rats / 中华普通外科杂志

Objective To study the effect of endothelial progenitor cells (EPCs) transplantation on CCl4 induced hepatic cirrhosis in rats. Methods Eight male SD rats were used as normal control. Thirty rats were induced liver cirrhosis by feeding with 25% CCl4/olive oil for 12 weeks, and then were subdivided into cirrhosis group (n = 10), EPCs transplanted group (n = 10) and saline control group (n = 10). EPCs were transplanted into the portal vein for 4 weeks in EPCs transplanted group. Rats in EPCs nontransplanted group were sacrificed at the beginning of the 12th week. Rats in EPCs transplanted group and saline control group were killed at the beginning of 16th week. Serum biochemical parameters were examined. The degree of liver cirrhosis was evaluated by Masson staining and by detecting the expression of α-SMA, Collagen Ⅲ and Ki67. Results The volumes of liver in cirrhosis group were twice as much as that in normal rats. 12 weeks after CCl4 administration, compared with saline control group, in EPCs transplanted group, hepatic activity index (HAI) ( F = 75. 062, P < 0. 01 ), the levels of ALT( F = 29. 942, P<0.05), AST(F=16.618,P<0.05) and TBIL(F=9.911 ,P<0.05) in serum decreased, the level of Alb ( F = 4. 944, P < 0. 05 ) and Ki67 ( F = 45. 966, P < 0. 01 ) was increased, the expression of α-SM A ( F = 7.86,P<0.05) and collagen Ⅲ (F = 135.787,P <0.01) decreased (P <0.05). Compared with untransplanted group, in EPCs transplanted group, the levels of ALT, AST and TBIL in serum were lower; In saline control group, the levels of ALT, AST and TBIL in serum were higher, the level of Alb and Ki67was lower, the expression of α-SMA and collagen Ⅲ were higher( P < 0. 05 ). Compared with normal rats, in saline control group, the levels of INR were higher (P < 0. 05 ). Conclusion EPCs transplantation improves hepatocye regeneration and ameliorates established hepatic cirrhosis.

Hepatitis B virus facilitates the expression of MMP-2 and TIMP-1 in hepatic stellate cells / 中华临床感染病杂志

Objective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.

Characteristics of hepatic progenitor cells activation in liver tissues of patients with hepatitis B cirrhosis / 中华临床感染病杂志

Objective To observe the characteristics of hepatic progenitor cells(HPCs)activation in liver tissues of patients with hepatitis B cirrhosis,and to investigate the relationship between the number of HPCs and HBV infection.Methods Cytokeratin 7(CK7)-was stained immunohistochemically in liver tissues of 16 patients with hepatitis B cirrhosis.HPCs and duetular reactions were quantitively analyzed.The expression of HBsAg and HBcAg were also detected to evaluate its relationship with HPCs activation.Results HPCs were extensively activated and marked duetular reactions can be observed in cirrhotic liver tissues.Tlle expression of HBsAg was positively correlated with HPCs activation.Conclusions HPCs are extensively activated in cirrhotic liver tissues,and HBV infection may facilitate its activation.

Effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with IL-2 or TNF-alpha / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2) or tumor necrosis factor-alpha (TNF-alpha).</p><p><b>METHODS</b>Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman's method. Using the isolated cells cultured and treated with IL-2 or TNF-alpha, we studied the effects of vitamin E on their proliferation and collagen synthesis through an (3)H-thymidine and (3)H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope.</p><p><b>RESULTS</b>Adding IL-2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-alpha, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-alpha to reproduce or synthesize collagen.</p><p><b>CONCLUSION</b>Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities.</p>

Transfusion of activated immune cells induced fluctuations of HCV titer without mutations in hepatitis C virus core and E2 region CTL epitopes / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the variations of HCV core and E2 region epitopes during transfusion of activated immune cells.</p><p><b>METHODS</b>Four patients receiving transfusion of activated immune cells were under continuously observation. HCV titers were measured by quantitive PCR. HCV core and E2 regions were cloned and sequences were analyzed by computer software.</p><p><b>RESULTS</b>During the follow-up the serum ALT levels and the HCV virus titers fluctuated greatly in each of these persons. After transfusion, no significant variations were observed in HCV core and E2 coding regions.</p><p><b>CONCLUSIONS</b>The alteration of host immune attacks could induce the fluctuations of HCV load without any mutations in the currently observed coding genes of the epitopes.</p>

Amplification of hepatitis C virus 5' untranslated region gene by RACE and its secondary structure analysis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To obtain very end full-length cDNA of hepatitis C virus (HCV) 5' untranslated region (5' UTR), and analyse its primary and secondary structure.</p><p><b>METHODS</b>By reverse transcription-nested polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP), a patient infected with genotype 2a HCV was found. Total RNA isolated from the serum as template, the cDNA of 5' noncoding region was amplified using rapid amplification of cDNA ends methods (RACE), the fragments were recombined by A-T clone strategy, the recombinants were confirmed by RFLP and PCR then sequenced. Secondary structures were analysed by RNA draw.</p><p><b>RESULTS</b>Very end full-length cDNA of 2a genotype HCV 5' UTR was obtained by RACE. In five clones obtained, three contained full-length 5' UTR cDNA, and A21G, G170A, T222C, T247C, C339T substitutions were found compared with HC-J6. he homologies with HCV-1,HC-J6,HC-C2, HC-J8 were 93.6%-94.4%, 92.1%-93.0%, 98.8%-99.7%, 96.2%-96.5%, respectively; however, the substitutions did not alter the secondary structure. Two out of five clones were deleted to have 53 and 144 bases at 5' terminus of HCV 5' UTR, respectively.</p><p><b>CONCLUSIONS</b>RACE is rapid and effective, works well to obtain very end of virus genome. With that, Authors obtained full-length cDNA of genotype 2a of HCV 5' UTR. There are genes deleted at 5' terminus circulated in hepatitis C patients.</p>

mRNA of MAGE genes as specific markers in detection of tumor cells in the peripheral blood of patients with hepatocellular carcinoma / 中华预防医学杂志

<p><b>OBJECTIVE</b>To detect tumor cells in the peripheral blood of patients with hepatocellular carcinoma (HCC) by using the mRNA of the MAGE-1 and MAGE-3 genes as specific tumor markers.</p><p><b>METHODS</b>Peripheral blood was obtained from 25 HCC patients and 20 healthy volunteers. The mRNA of the MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMCs) was detected by nested RT-PCR. The MAGE-1 and MAGE-3 transcripts in the tumor tissues of these HCC patients were also detected by RT-PCR.</p><p><b>RESULTS</b>Of the 25 HCC patients, MAGE-1 and MAGE-3 mRNA were positive in 44% (11/25) and 36% (9/25) of PBMCs respectively, and in 68% (17/25) and 56% (14/25) of HCC tissues respectively. In the PBMCs of the 25 HCC patients, 16 (64%) samples were detected to express at least one type of MAGE mRNA. MAGE mRNA were not detected in the PBMCs from the patients whose tumors did not express the MAGE genes, nor in the PBMCs from the 20 healthy donors. The positive rate of MAGE mRNA in the PBMCs was closely correlated with the TNM stages and the diameter of tumors, but there was no correlation between the positive rate of MAGE mRNA in PBMCs and tumor differentiation degree or serum alpha-FP level. Of 9 HCC patients whose serum alpha-FP was normal or slightly elevated (< 50 ng/ml), 6 were MAGE-1 and/or MAGE-3 mRNA positive in their PBMCs.</p><p><b>CONCLUSION</b>MAGE-1 and MAGE-3 mRNA could be specifically detected with high percentage in the PBMCs of HCC patients by our method. They can be used as specific tumor markers for the detection of the circulating HCC cells, and the detection results may be helpful to evaluate the prognosis of HCC patients.</p>

Comparison of Serum-Free Medium AIMV with Standard Serum-Containing Medium in Culturing Cells Activated with IFN-?,IL-2 and Anti-CD3 / 中国肿瘤生物治疗杂志

Objective: To compare serum-free medium AIMV with standard serum-containing medium in culturing cells activated with IFN-?,IL-2 and anti-CD3. Methods: Peripheral blood mononuclear cells (PBMC) separated from donors were cultured with IFN-?,IL-2 and anti-CD3 in serum-free medium AIMV or in standard serum-containing medium separately. Proliferation, phenotypes and cytokine secretion of cells cultured in the two different media were compared. The inhibitory effects on hepatitis B virus replication in vivo were observed after transfusion of cells cultured in different media. Results: Compared with cells cultured in standard serum-containing medium, cells cultured in serum-free medium AIMV could proliferate well enough to be applied to immunotherapy; More percentage of cells cultured in AIMV expressed CD25; Cells cultured in serum-free medium secreted more IFN-?; After transfusion of cells cultured in AIMV, hepatitis B replication could be inhibited more evidently. Conclusion: Serum-free medium AIMV was better than serum-containing medium in culturing cells for immunotherapy.

The immune response of CD8~+T cells from subjects of different HLA-A~*02 subtype detected by intracellular cytokine staining / 中国免疫学杂志

Objective:Intracellular cytokine staining(ICCS) is applied to the detection of the immune response of peripheral blood mononuclear cells (PBMC)from subjects of different HLA A 02 supertype to influenza A matrix peptide.Methods:HLA A 02 subtype were identified by the way of SSP methods;PBMC from subjects with HLA A 02 were cocultured with influenza A matrix peptide (IMP) which was HLA A*0201 restricted CTL epitope, secreting cytokine such as IL 2, IFN ?,TNF ? were measured by ICCS.Results:PBMC from HLA A 02 positive subjects had memory immune response to IMP.Conclusion:①The method of ICCS can be used for quantitive detecting T cell activating status specificly;②the memory immune response to IMP58 66 peptide exists in all of the HLA A 02 positive persons;③among different HLA A 02 subtype, there are cross reactivity to HLA A 0201restricted peptide.