BACKGROUND:Ferroptosis is a regulated, iron-dependent form of cell death triggered by toxic accumulation of lipid peroxides on cell membranes. Although it is established that glycosphingoid metabolites can activate the ferroptosis signaling pathway, whether they are involved in regulating endoplasmic reticulum stress (ERS)-ferroptosis in immune-mediated liver injury requires further investigation. METHODS:This study used the GCS inhibitor GENZ-123346 (GENZ) to treat ConA-induced immune liver injury in mice. Inflammatory factor levels and liver histology were analyzed to evaluate GENZ's effects. Additionally, the antioxidant and lipid peroxidation capacity of GENZ was tested in vivo and in vitro. Based on the regulation of NFE2-related factor 2 (NRF2) signaling pathway expression (including si-NRF2, NRF2 inducer sulforaphane (SFN), and inhibitor ML385), the regulatory effect of GENZ on ERS-ferroptosis was investigated. RESULTS:GENZ administration improved the survival rate of mice with immune-mediated liver injury, decreased serum transaminase activity, inhibited inflammatory factor production, reduced hepatocyte ferroptosis. In in vitro experiments, ferroptosis in AML12 cells was accompanied by the activation of ERS-related proteins Glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) and mild alterations in endoplasmic reticulum ultrastructure. Inhibition of ERS by 4-phenylbutyric acid (4-PBA) attenuated ferroptosis in AML12 cells. However, when the cells were treated with the ferroptosis inducer RSL3 in combination with the ERS inducer thapsigargin (Tg), GENZ could not effectively suppress ferroptosis. Silencing NRF2 exacerbated cellular ferroptosis and ERS while diminishing the expression of downstream effectors heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1). Conversely, SFN treatment alleviated these effects and upregulated NRF2 signaling, increasing HO-1 and NQO1 expression. Furthermore, ML385 administration reversed the hepatoprotective and antioxidant effects of GENZ on immune-mediated liver injury in mice. CONCLUSIONS:GCS inhibition exerts hepatoprotective effects by modulating the NRF2 antioxidant pathway to suppress ERS and hepatocyte ferroptosis, revealing a novel role of glycosphingolipid metabolism in immune-mediated liver injury and programmed hepatocyte death.

Background: Although Th17 cells and regulatory T cells play critical roles in autoimmune hepatitis, the balance between them in acute and chronic immune hepatitis is not well-studied. The objective of this study was to explore the role of Th17/Treg balance in autoimmune hepatitis. Methods: Thirty female Wistar rats were randomly divided into the following three groups: acute model (AC), chronic model (CC), and healthy control (HC). Aspartate aminotransferase, alanine aminotransferase, and bilirubin levels were measured. Peripheral blood interleukin (IL)-17, IL-6, IL-10, and transforming growth factor-beta cytokine levels were also assessed. Flow cytometry was used to detect changes in the Th17 cell and Treg cell frequency, and the Th17/Treg ratio was calculated. Results: The frequency of Th17 cells and Treg cells were both increased (p<0.05) and the Th17/Treg ratio was higher in the AC group compared with those in the HC group (p<0.05). Th17 cell levels were significantly lower and Treg cell levels were significantly higher in the CC group, compared with those in the AC group (p<0.05). In addition, the Th17/Treg ratio was lower in the CC group compared with that in the HC group (p<0.05). Conclusion: The Th17/Treg ratio was increased in the acute immune liver injury model and decreased in the chronic liver injury model. Th17/Treg imbalance may play a critical role in liver injury occurrence and development.

布鲁菌病是一种常见的人畜共患疾病,发病范围广泛.布鲁菌病骨关节受累患病率达10％～85％,其中脊柱关节和骶髂关节更为常见.布鲁菌病骨关节损害的临床表现多样,需要与腰背部疼痛相关疾病鉴别,早期正确诊断和治疗是关键.

Liver biopsy is the gold standard for staging liver fibrosis, but it has numerous drawbacks, mainly associated with bleeding and bile fistula risks. A number of non-invasive techniques have been investigated, but they all have their own disadvantages. To avoid the risks mentioned above and to improve the diagnostic value, we still need to search for a more accurate non-invasive method to evaluate the degree of liver fibrosis. This study aimed to evaluate the diagnostic performance of FibroTouch versus other non-invasive fibrosis indexes in hepatic fibrosis of different aetiologies. This study retrospectively enrolled 227 patients with chronic hepatic liver disease admitted to the first hospital of Lanzhou University from 2017 to 2020. Liver biopsy was performed in all of the patients, and their biochemical indicators were all tested. Non-invasive indexes including the fibrosis index based on four factors (FIB-4), the aminotransferase-to-platelet ratio index (APRI), and the gamma-glutamyl transpeptidase-to-platelet ratio index (GPRI) were all calculated. Transient elastography was performed using FibroTouch. The correlation between FibroTouch and the pathology of liver fibrosis was significantly higher than that between the non-invasive fibrosis indexes and the biopsy results (r = 0.771, p < 0.05). The area under the receiver operating curve (AUC) of FibroTouch was significantly higher than that of FIB-4, APRI, and GPRI for the diagnosis of significant fibrosis (≥ S2 fibrosis stage), advanced fibrosis (≥ S3 fibrosis stage), and cirrhosis (= S4 fibrosis stage) (p < 0.05). The patients were grouped according to different aetiologies. The diagnostic value of FibroTouch had much higher credibility in different fibrosis stages for different causes compared with other non-invasive indexes. The AUC of FibroTouch showed both higher specificity and higher sensitivity than FIB-4, APRI, and GPRI for different liver fibrosis stages with different aetiologies. FibroTouch demonstrates the highest diagnostic value for liver fibrosis and cirrhosis among non-invasive methods, showing better results than FIB-4, APRI, and GPRI, and surpassed only by liver biopsy. FibroTouch is reliable in assessing liver fibrosis with different aetiologies.

Objective: This study aimed to explore the effect of miR-96-5p targeting of PRC1 (protein regulator of cytokinesis 1) gene on proliferation, apoptosis and invasion of hepatoma HepG2 cells. Methods: LO2 and HepG2 cells were cultured in vitro for the study. The targeted relationship between miR-96-5p and PRC1 was verified by dual-luciferase reporter gene assay. HepG2 cells were separately transfected within seven groups including; Control group, mimic NC group, miR-96-5p mimic group, miR-96-5p inhibitor group, oe NC group, oe PRC1 group, and miR-96-5p mimic + oe PRC1 group. Expression of miR-96-5p and PRC1 were detected by quantitative reverse transcriptase PCR and western blot. Proliferation activity was detected by 5-ethynyl-2'-deoxyuridine (EdU); cell cycle and apoptosis were determined by flow cytometry. Cell metastasis and invasion were assessed by Transwell assay. The effects of the overexpression of PRC1 or miR-361-5p on tumor growth were evaluated through tumorigenesis experiment in nude mice. Results: Compared with normal liver LO2 cells, miR-96-5p expression was down-regulated and PRC1 expression was up-regulated in hepatoma HepG2 cells. Dual-luciferase reporter gene assay showed that there is a targeted relationship between miR-96-5p and PRC1. miR-96-5p targeted the 3' untranslated region (3'UTR) of the PRC1 gene and down-regulated its expression. Overexpression of miR-96-5p could inhibit proliferation, migration and invasion of HepG2 cells, suppress tumor growth and promote cell apoptosis. Inversely, overexpression of PRC1 elicited an opposite response. Meanwhile, overexpression of both miR-96-5p and PRC1 in miR-96-5p mimic + oe PRC1 group, compared with miR-96-5p mimic group, presented an enhancement of proliferation, migration and invasion of HepG2 cells as well as inhibition on apoptosis. While compared with oe PRC1 group, HEPG 2 cells in miR-96-5p mimic + oe PRC1 group presented with significantly opposite effects. Conclusion: miR-96-5p targeting of PRC1 gene inhibits proliferation and invasion of hepatoma HepG2 cells and promotes their apoptosis.

Background Chronic hepatitis B virus (HBV) infection-reduced liver functions are associated with intestinal microbial community dissimilarity. This study aimed to investigate the microbial community dissimilarity in patients with different grades of HBV-related liver cirrhosis. Results Serum endotoxin was increased with Child–Pugh (CP) class (A, B, and C). Veillonellaceae and Lachnospiraceae families were reduced in patients compared with controls. Megamonas and Veillonella genus was reduced and increased in patients compared with controls, respectively, especially in CPB and CPC groups. Correlation analysis showed that endotoxin content was significantly correlated with alcohol consumption (95% CI 0.100, 0.493), CP class (95% CI 0.289, 0.687) and Lachnospiraceae family level (95% CI − 0.539, − 0.122). Firmicutes/Bacteroidetes ratio was correlated with the level of Lachnospiraceae family (95% CI 0.013, 0.481), Veillonellaceae family (95% CI 0.284, 0.696), Megamonas genus (95% CI 0.101, 0.518) and Veillonella genus (95% CI 0.134, 0.545). All aforementioned bacteria were independent risk or protective factors for hepatitis. Alcohol consumption changed microbial community. Conclusions Our study demonstrated that elevated Firmicutes/Bacteroidetes ratio, reduced Megamonas genus level and increased Veillonella genus level were indicators for HBV-related liver cirrhosis. Alcohol-related pathogenesis was associated with the changed microbial community.

Objective To investigate the changes in procoagulant and anticoagulant factors and routine coagulation markers in patients with hepatitis B cirrhosis and their clinical significance. Methods A total of 105 patients with hepatitis B cirrhosis who were admitted to Department of Infectious Diseases in The First Hospital of Lanzhou University from January 2017 to October 2018 were enrolled, and according to the Child-Pugh class, these patients were divided into group A (42 patients with Child-Pugh class A cirrhosis), group B (39 patients with Child-Pugh class B cirrhosis), and group C (24 patients with Child-Pugh class C cirrhosis). Routine coagulation markers including prothrombin time (PT) and activated partial prothrombin time (APTT), blood coagulation factors (Ⅱ,Ⅴ, Ⅶ, Ⅷ, and Ⅺ), anti-thrombin (AT), protein C (PC), and free protein S (FPS) were measured for all patients. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups; a Spearman correlation analysis was also performed. Results There were significant differences in PT and APTT between the three groups (F=55.11 and 12.09, both P＜0.001); group C had significantly higher PT and APTT than groups A and B, and group B had significantly higher PT and APTT than group A (all P＜0.05). There were significant differences in the activities of blood coagulation factors Ⅱ, Ⅴ, Ⅶ, and XI between the three groups (F=32.52, 14.77, 38.88, and 9.24, all P＜0.001); group C had significantly lower activities of these coagulation factors than groups B and A, and group B had significantly lower activities than group A (all P＜0.05). There was a significant difference in the activity of blood coagulation factor Ⅷ between the three groups (F=4.44, P＜0.05); group C had a significantly higher activity than groups A and B (P＜0.05), while there was no significant difference between group A and group B (P＞0.05). There were significant differences in the activities of AT, PC, and FPS between the three groups (F=25.90, 30.46, and 15.58, all P＜0.001); group C had significantly lower activities than groups B and A, and group B had significantly lower activities than group A (all P＜0.05). The correlation analysis showed that Child-Pugh class was negatively correlated with blood coagulation factors Ⅱ, Ⅴ, Ⅶ, and Ⅺ and anticoagulant factors PC, FPS, and AT (r=-0.687, -0.460, -0.706, -0.426, -0.723, -0.646, and -0.468, all P＜0.001), and Child-Pugh class had the strongest correlation with blood coagulation factors Ⅱ and Ⅶ and the anticoagulant factor PC. Blood coagulation factors II and Ⅶ were positively correlated with the anticoagulant factor PC (r=0.851 and 0.745, both P＜0.001). Conclusion Patients with hepatitis B cirrhosis have reductions in both procoagulant and anticoagulant factors, and there is a significant correlation between them. An unstable balance may form in the body, which is not considered a conventional hypocoagulable state.

The effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the proliferation of hepatic stellate cells (HSCs) is largely unknown. The purpose of this study was to explore the mechanism of action of hUC-MSCs on the proliferation of HSCs in vitro. The upper and lower double-cell co-culture system was established between hUC-MSCs and HSCs in the experimental group. HSCs were cultured alone as a negative control group. Cell proliferation and apoptosis were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. Cell supernatants were harvested to determine the concentration of transforming growth factor-1 (TGF-1) by ELISA. mRNA and protein of TGF-1, Smad3 and Smad7 in HSCs were determined by reverse transcription-polymerase chain reaction and western blotting, respectively. In the co-culture group, the proliferation of HSCs was significantly inhibited compared with the negative control group at 24 and 48 h (p<0.05). Apoptosis of HSCs in the co-culture group increased compared with that in the negative control group, which was more obvious at 48 h (p<0.05). The concentration of TGF-1 in the co-culture group was significantly lower than in the HSCs cultured alone (p<0.05). After HSCs were co-cultured with hUC-MSCs for 48 h, expression of TGF-1 and Smad3 mRNA and protein was reduced and expression of Smad7 mRNA and protein was increased compared with the negative control group (p<0.05). hUC-MSCs inhibited proliferation of HSCs, possibly through inhibiting TGF-1 and Smad3 expression and increasing Smad7 protein expression.

Objective To explore the application of the teaching method combining case-based learning(CBL) with problem-based learning(PBL) in clinical teaching of infectious diseases. Methods A total of 60 students for infectious diseases on clinical training during the february to july of 2017 were divided into PBL combined with CBL group and LBL group.Evaluation of the teaching effect of two kinds of teaching methods. Results The results of Theoretical examination,case analysis and total score in PBL+CBL group were higher than those of LBL group. Difference is statistically significant(P<0.05).Meanwhile,knowledge understanding, clinical practice interesting,communication ability between doctors and patients, capability of consulting documents and satisfaction with teaching methods were better than those in LBL group (P<0.05). Conclusion The PBL combined with CBL teaching method can improve the teaching effect of clinical practice of infectious diseases,which is worthy of further popularization and application.

It has been confirmed that the inhibitors of poly ADP-ribose polymerF(<^>9ase-1 (PARP-1) can inhibit the proliferation, apoptosis and invasion of tumor cells. However, the effects of inhibitors of PARP-1 on hepatocellular carcinoma remain to be elucidated. The aim of the present study was to investigate the effect of three types of PARP-1 inhibitor on the proliferation, apoptosis and migration of hepatocellular carcinoma in vitro. An MTT assay was performed to detect the proliferation of HepG2 cells following treatment with the PARP-1 inhibitors, AG014699, BSI-201 and AZD-2281. Flow cytometry was used to detect the apoptosis of HepG2 cells, Western blot analysis was used to detect the protein expression of Casepase-3, Casepase-8, B-cell lymphoma 2 (Bcl-2) -associated X protein (Bax), Bcl-2, phosphatase and tensin homolog (PTEN), tissue inhibitor of metalloproteinase (TIMP) 3 and matrix metalloprotease (MMP) 3. A Transwell assay was performed to detect the migration of HepG2 cells. The results showed that AG014699, BSI-201 and AZD-2281 had an inhibitory effect on the proliferation of HepG2 cells in a time-and concentration-dependent manner. AG014699 at concentrations of 10, 30 and 50 mu mol/l, and BSI-201 at concentrations of 20, 40 and 60 mu mol/l induced the apoptosis of HepG2 cells, and the apoptotic rates were particularly high at 48 h (31, vs. 0.01%; P < 0.01 and 24.12, vs. 0.03%, respectively; P < 0.01). The protein expression levels of Caspase 3, Caspase 8, Bax, PTEN and TIMP 3 increased with increasing drug concentrations, whereas the protein levels of Bcl-2 and MMP3 decreased with increasing drug concentrations, and were significantly different compared with those in the control group (P < 0.01). In conclusion, AG014699, BSI-201 and AZD-2281 inhibitors of PARP-1 significantly inhibited the proliferation of HepG2 cells, however, AG014699 and BSI-201 demonstrated more sensitivity, induced apoptosis and inhibited migration of the hepatocellular carcinoma cells, which may be associated with alterations of the apoptosis signaling pathway and the expression of proteins associated with migration.

Objective It has been confirmed that the inhibitors of poly ADP-ribose polymerase-1 (PARP-1) affect the proliferation,apoptosis and invasion of tumor cells.However,the impact of the inhibitors of PARP-1 on hepatoma cells remains unclear.The purpose of this study was to investigate the effect of three kinds of PARP-1 inhibitors on the proliferation,apoptosis and migration of hepatocellular carcinoma cells in vitro.Methods MTT assay was performed to detect the proliferation of HepG2 cells after treatment with PARP-1 inhibitors,AG014699,BSI-201,and AZD-2281.Thereafter,AG014699 and BSI-201,which had the most suppressive effect,were used to treat HepG2 cells.Western blot assay was used to detect the protein expression of asepase3,casepase8,Bax,Bcl-2,PTEN,tissue inhibitor of metalloproteinase 3 (TIMP3),and matrix metalloprotease 3 (MMP3).Transwell assay was performed to detect the effect of AG014699 and BSI-201 on the migration of HepG2 cells.Results AG014699,BSI-201 and AZD-2281 all could inhibit the proliferation of HepG2 cells in a time-and concentration-dependent manner.The protein expression levels of caspase3,caspase8,Bax,PTEN,TIMP3 increased and those of Bcl-2 and MMP3 decreased,with the increasing of the drug concentration,in HepG2 cells treated for 48 h,which showed a significant difference as compared with the control group (P<0.01).Conclusion All PARP-1 inhibitors AG014699,BSI-201,and AZD-2281 could significantly inhibit the proliferation of HepG2 cells,with AG014699 and BSI-201 having better sensitivity.AG014699 and BSI-201 could significantly induce the apoptosis and inhibit the migration of hepatoma cells,which may be related to the alteration of apoptosis-related signaling pathway and the expression of migration-related proteins.

Objective To explore the influencing factors for chronic hepatitis C (CHC) with thyroid dysfunction (TD) in untreated Chinese patients and provide evidence for clinical individualized treatment. Methods One thousand and twelve untreated CHC patients were collected nationwide in China. Thyroid function and associated influencing factors (region, age, gender, and hepatitis C virus (HCV) RNA replication level) in the patients were investigated. The relationships between the influencing factors and CHC with TD were analyzed. Between-group comparison of categorical data was performed by χ2 test and Fisher′s exact test. Results There were geographical differences between different types of CHC with TD. Across different regions, the incidence of TD was highest in north and northwest China, i.e., 28.3% and 26.5%, respectively. Subclinical hypothyroidism was the most common type of TD, accounting for 58.8% of the total TD cases. Middle-aged patients were most common among the cases of CHC with TD (44.0%), who had a significantly higher incidence of hypothyroidism than other age groups (χ2=10.10、6.17, P=0.001、0.013). Females with CHC had a significantly higher incidence of TD than male patients (58.9% vs. 41.1%, χ2=13.1, P=0.00). Although a high HCV RNA replication level was most common in Chinese patients with CHC, this factor had little influence on TD. Conclusion In China, CHC with TD is influenced by geographic distribution, gender, and age, but less associated with HCV RNA replication level.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are considered to be a potential therapy for end-stage liver disease. However, the therapeutic mechanism of BM-MSCs remains unclear. The aim of the current study was to investigate the role of paracrine signaling in BM‑MSCs in liver cirrhosis in vitro. Human BM‑MSCs and hepatic stellate cells (HSCs) were cultured using a vertical double cell co‑culture system. Groups were divided into HSCs alone (control group) and the co‑culture system of BM‑MSCs with HSCs (experimental group). HSC morphology was observed by inverted phase contrast microscopy. The proliferative capacity of HSCs was measured with the MTT assay and flow cytometry. Hoechst staining was performed to examine the apoptosis of HSCs. Transforming growth factor (TGF)‑β1 and Smad7 mRNA expression were detected by reverse transcription‑quantitative polymerase chain reaction and western blotting. BM‑MSCs did not inhibit the proliferation of HSCs at 24 h, however significantly inhibited the proliferation of HSCs at 48 and 72 h. BM‑MSCs additionally induced the apoptosis of HSCs at 48 h. The concentration of TGF‑β1 in the supernatant at 24 h and 48 h in the co‑cultured system was observed to be significantly lower than in the control group (P<0.05). The level of TGF‑β1 mRNA in the experimental group at 48 h was significantly lower than the control group, however Smad7 mRNA levels were significantly greater than in the control group. Additionally, TGF‑β1 protein levels were significantly lower than in the control group, however levels of Smad7 were greater than the control group. It was concluded that BM‑MSCs are able to inhibit the proliferation and promote the apoptosis of HSCs. In addition, the mechanism may be associated with inhibition of the TGF-β1/Smad pathway in HSCs.

OBJECTIVE:To investigate the distribution of HCV genotypes in Chinese Han population with chronic hepatitis C (CHC).METHODS:This randomized multicenter study included 1 014 CHC patients from 28 hospitals in different regions of China. SPSS 20.0 was applied to analyze the relationship among region, HCV genotype, gender and the replication level of HCV-RNA.RESULTS:HCV 1 genotype (56.80%) was the most common genotype. The majority of CHC patients were of genotype 1, 2, 3, 6 in the order of frequency, except those in southwestern, southern and central China. HCV 1, 2, 3, 6 genotypes were most common among male patients in southern China; among female patients in northern China; among male patients in northern and northwestern China and among male patients in northwestern China, respectively (all P <0.05). There was no statistical significance between different genders in other regions. The high viral load was more common than the low viral load among HCV 1, 2, 3, 6 genotype-infected patients.CONCLUSION:There are different distributions of HCV genotypes among the different regions. In addition, HCV genotypes are correlated with gender and HCV-RNA load.

OBJECTIVE:To investigate the association of hepatitis C virus (HCV) genotype with glycolipids iron metabolism in Gansu Han population.METHODS:The genotypes of HCV 1b type and 2a type were detected in Gansu Han HCV carriers. The Glu, Insulin, CHOL, TG, UIBC, TRF, TIBC, SF, Serum Iron, AST, ALT, TBil, IBil, DBil, ALP, GGT were measured and compared between patients with different HCV genotypes.RESULTS:There were 84 cases with HCV1b type and 136 cases with 2a type. There were significant differences in TG, ALT, TRF, TIBC between 1b type and 2a type genotype HCV carriers.CONCLUSION:The 2a type HCV carriers may be more inclined to develop hyperlipidemia and liver damage, and 1b type HCV carriers are likely to have iron metabolism defect.

ObjectiveTo investigate the genotypes of hepatitis C virus (HCV) among the hepatitis C patients of Han nationality in Gansu, China, as well as their possible routes of transmission and relationship with clinical indices. MethodsThe complete data regarding hepatitis C, including sex, age, possible routes of transmission, HCV RNA level, and biochemical parameters, were collected from 220 hepatitis C patients of Han nationality in Gansu. Continuous data were analyzed by t test and rank sum test; categorical data were analyzed by chi-square test; multivariate logistic regression analysis was used to investigate the relationship between HCV genotypes and clinical indices. ResultsHCV genotype 1b was detected in 84 (38%) of all patients, and genotype 2a in 136 (62%) of these patients. The univariate analysis of 11 factors including age, sex, route of transmission, HCV RNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), indirect bilirubin (IBil), direct bilirubin (DBil), alanine phosphatase (ALP), and gamma-glutamyl transpeptidase (GGT) showed that the route of transmission, ALT, and HCV RNA were correlated with HCV genotypes (χ2 = 23.947, P＜0.001; Z=-3.349, P=0.001; t=-2.325, P=0.021), but age, sex, AST, TBil, IBbil, DBil, ALP, and GGT were not correlated with HCV genotypes (P＞0.05). The multivariate logistic regression analysis showed that the above three factors were correlated with HCV genotypes (χ2=3.993, P=0.046; χ2=9.308, P=0.002; χ2=11.652, P=0.001). The number of genotype 2a patients was larger than that of genotype 1b patients among patients with high HCV RNA levels (≥106 IU/ml) and those with high ALT levels (＞49 U/L). HCV genotype 1b was more likely to be transmitted by blood transfusion and oral treatment compared with other routes. ConclusionGenotype 1b and genotype 2a are the main genotypes of HCV among the hepatitis C patients of Han nationality in Gansu, and genotype 2a is more common. Genotype 2a patients are more prone to high HCV RNA level and liver damage. In Gansu, HCV is transmitted mainly by blood transfusion, followed by oral treatment, and genotype 1b is more likely than genotype 2a to be transmitted by the two routes.