Objective:To explore the influences of FibroScan detection on the pregnancy outcomes of pregnant rats transfected with hepatitis B virus and the growth of their offspring rats.Methods:Sixty SPF-grade SD rats with a male-to-female ratio of 5∶1 were selected，randomly divided the female rats into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group，and the rats in the mid-pregnancy and late-pregnancy groups were subjected to FibroScan testing on the 10th and 15th days of gestation respectively. The number of offspring rats in each group was randomly reduced to 10 on the 3rd day after delivery，and the offspring rats were divided into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group. The pregnancy outcome levels of pregnant rats were observed by adopting a stratified analysis strategy，including differences in weight changes，number of deliveries，fetal rat outcomes，lactation conditions，litter size，and litter weight. The growth levels of offspring rats，including differences in weight，body length，tail length，and the time of fur growth，tooth eruption，eye opening，and ear standing on the 21st day were observed，and the survival rate of the offspring rats was compared.Results:With regard to the pregnancy outcome levels of the pregnant rats，there were no statistically significant differences in weight changes during pregnancy among the five groups of pregnant rats［ F（4，45）=2.627，Adjusted P=0.222］. There were also no statistically significant differences in the number of deliveries，fetal rat outcomes，and lactation conditions（Adjusted P=1.000）. The number of deliveries［ F（4，21.095）=2.280，Adjusted P=0.222］and litter weight［ F（4，20.128）=2.159，Adjusted P=0.222］showed no statistically significant differences. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. In terms of the growth levels of the offspring rats，among the five groups of offspring rats，there were no statistically significant differences in body weight on the 21st day［H（4）=11.623，Adjusted P=0.135］，body length on the 21st day［H（4）=10.962，Adjusted P=0.135］，and tail length on the 21st day［H（4）=9.126，Adjusted P=0.058］. Besides，the differences in the time of fur growth，tooth eruption，eye opening，and ear standing［H（4）=0.000，Adjust P=1.000］showed no statistical significance. The survival rate on the 7th day，14th day，and 21st day was 100%，with no significant differences（Adjusted P=1.000）. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. Conclusion:FibroScan detection had no significant influences on the pregnancy outcomes of pregnant rats with hepatitis B or on the growth of their offspring rats in multiple stages，dimensions，and indicators，indicating that FibroScan detection is safe for pregnant rats. This research provides an animal experimental basis for the safe use of FibroScan in pregnant women with chronic HBV infection.

Objective:To establish a new method and platform for screening, identifying, and exploring a new strategy for anti-hepatitis B immunotherapy based on hepatitis B virus (HBV)-specific TCR.Methods:Peripheral blood mononuclear cells were isolated from patients with acute hepatitis B. CD3 +CD8 +CD137 +T single cells were sorted out after stimulation with the HBsAg peptide library. The α and β chains in TCRs of single cells were amplified by PCR. TCR double-chain pairing and lentiviral packaging were performed through high-throughput sequencing. Re-infected Jurkat-76-NFAT-GFP cells and the cell lines stably expressing TCR were screened. HBsAg peptide library and immortalized B lymphocytes co-cultured with J76N-TCR were used to screen HBsAg-specific TCRs. K562 cell lines stably expressing HLA-A*24:02 were established to determine epitope peptide by screening A*24:02-restricted TCR. The screened TCRs were replaced with mouse C regions and packaged with lentiviruses. Functional validation was performed on healthy human CD4 +T and CD8 +T lymphocytes following infection. Results:Stable TCR-expressing cell lines were successfully prepared based on single-cell TCRαβ double-chain amplification and pairing technology. Twenty-one TCRs were screened using immortalized B lymphocytes, resulting in nine possible HLA-A*24:02-restricted HBsAg-specific TCRs. Further screening with K562-A2402 resulted in six A*24:02-restricted HBsAg-specific TCRs with identically recognized epitope peptide. The functional determination of the two TCR clones revealed their specific recognition function for target cells expressing HBsAg.Conclusion:HLA-A*24:02-restricted HBsAg-specific TCR with recognition function for target cells expressing HBsAg was successfully obtained based on the new experimental technology system, laying an important foundation for further exploration of antiviral immunotherapy based on HBV-specific TCR.

Purpose To investigate the patterns of change in the topological organization of individual level gray matter morphological networks in children with autism spectrum disorder(ASD).Materials and Methods The clinical and high-resolution T1 structural image data of 52 ASD children and 69 typical developing children from Autism Brain Imaging Data Exchange public database were selected.The individual gray matter structural networks were constructed by calculating the similarities of regional gray matter morphology.The global and local topological metrics of individual gray matter networks were analyzed using a graph theory method and compared between the two groups.Partial correlation analysis was used to evaluate the correlation between topological metrics showing significant between group differences and clinical characteristics in the ASD group.Results The verbal IQ was significantly lower in the ASD group compared with the typical developing group(t=-3.151,P=0.002),and no significant differences were found in full scale IQ and performance IQ between the two groups(P>0.05).Compared with the typical developing group,the ASD group showed significantly higher global efficiency(P=0.014),lower clustering coefficient(P=0.044)and characteristic path length(P=0.020)of gray matter structural networks.In the ASD group,abnormal nodal centralities were found in multiple brain regions(P<0.05,false discovery rate corrected),including the bilateral dorsolateral superior frontal gyrus,bilateral precuneus,bilateral caudate nucleus,right superior temporal gyrus,left middle temporal gyrus and bilateral anterior cingulate gyrus.No significant associations were found between the abnormal network topological metrics and IQ scores in the ASD group(r=0.069-0.187,all P>0.05).Conclusion Altered topological organization of individualized gray matter morphological networks is found in children with ASD,as characterized by decreased local segregation and increased global integration of networks.The areas with abnormal nodal centralities in children with ASD are mainly located at the brain regions involved in emotional processing,social and high-order cognitive functions.

Purpose To investigate the patterns of change in the topological organization of individual level gray matter morphological networks in children with autism spectrum disorder(ASD).Materials and Methods The clinical and high-resolution T1 structural image data of 52 ASD children and 69 typical developing children from Autism Brain Imaging Data Exchange public database were selected.The individual gray matter structural networks were constructed by calculating the similarities of regional gray matter morphology.The global and local topological metrics of individual gray matter networks were analyzed using a graph theory method and compared between the two groups.Partial correlation analysis was used to evaluate the correlation between topological metrics showing significant between group differences and clinical characteristics in the ASD group.Results The verbal IQ was significantly lower in the ASD group compared with the typical developing group(t=-3.151,P=0.002),and no significant differences were found in full scale IQ and performance IQ between the two groups(P>0.05).Compared with the typical developing group,the ASD group showed significantly higher global efficiency(P=0.014),lower clustering coefficient(P=0.044)and characteristic path length(P=0.020)of gray matter structural networks.In the ASD group,abnormal nodal centralities were found in multiple brain regions(P<0.05,false discovery rate corrected),including the bilateral dorsolateral superior frontal gyrus,bilateral precuneus,bilateral caudate nucleus,right superior temporal gyrus,left middle temporal gyrus and bilateral anterior cingulate gyrus.No significant associations were found between the abnormal network topological metrics and IQ scores in the ASD group(r=0.069-0.187,all P>0.05).Conclusion Altered topological organization of individualized gray matter morphological networks is found in children with ASD,as characterized by decreased local segregation and increased global integration of networks.The areas with abnormal nodal centralities in children with ASD are mainly located at the brain regions involved in emotional processing,social and high-order cognitive functions.

Objective:To explore the influences of FibroScan detection on the pregnancy outcomes of pregnant rats transfected with hepatitis B virus and the growth of their offspring rats.Methods:Sixty SPF-grade SD rats with a male-to-female ratio of 5∶1 were selected，randomly divided the female rats into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group，and the rats in the mid-pregnancy and late-pregnancy groups were subjected to FibroScan testing on the 10th and 15th days of gestation respectively. The number of offspring rats in each group was randomly reduced to 10 on the 3rd day after delivery，and the offspring rats were divided into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group. The pregnancy outcome levels of pregnant rats were observed by adopting a stratified analysis strategy，including differences in weight changes，number of deliveries，fetal rat outcomes，lactation conditions，litter size，and litter weight. The growth levels of offspring rats，including differences in weight，body length，tail length，and the time of fur growth，tooth eruption，eye opening，and ear standing on the 21st day were observed，and the survival rate of the offspring rats was compared.Results:With regard to the pregnancy outcome levels of the pregnant rats，there were no statistically significant differences in weight changes during pregnancy among the five groups of pregnant rats［ F（4，45）=2.627，Adjusted P=0.222］. There were also no statistically significant differences in the number of deliveries，fetal rat outcomes，and lactation conditions（Adjusted P=1.000）. The number of deliveries［ F（4，21.095）=2.280，Adjusted P=0.222］and litter weight［ F（4，20.128）=2.159，Adjusted P=0.222］showed no statistically significant differences. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. In terms of the growth levels of the offspring rats，among the five groups of offspring rats，there were no statistically significant differences in body weight on the 21st day［H（4）=11.623，Adjusted P=0.135］，body length on the 21st day［H（4）=10.962，Adjusted P=0.135］，and tail length on the 21st day［H（4）=9.126，Adjusted P=0.058］. Besides，the differences in the time of fur growth，tooth eruption，eye opening，and ear standing［H（4）=0.000，Adjust P=1.000］showed no statistical significance. The survival rate on the 7th day，14th day，and 21st day was 100%，with no significant differences（Adjusted P=1.000）. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. Conclusion:FibroScan detection had no significant influences on the pregnancy outcomes of pregnant rats with hepatitis B or on the growth of their offspring rats in multiple stages，dimensions，and indicators，indicating that FibroScan detection is safe for pregnant rats. This research provides an animal experimental basis for the safe use of FibroScan in pregnant women with chronic HBV infection.

Objective:To establish a new method and platform for screening, identifying, and exploring a new strategy for anti-hepatitis B immunotherapy based on hepatitis B virus (HBV)-specific TCR.Methods:Peripheral blood mononuclear cells were isolated from patients with acute hepatitis B. CD3 +CD8 +CD137 +T single cells were sorted out after stimulation with the HBsAg peptide library. The α and β chains in TCRs of single cells were amplified by PCR. TCR double-chain pairing and lentiviral packaging were performed through high-throughput sequencing. Re-infected Jurkat-76-NFAT-GFP cells and the cell lines stably expressing TCR were screened. HBsAg peptide library and immortalized B lymphocytes co-cultured with J76N-TCR were used to screen HBsAg-specific TCRs. K562 cell lines stably expressing HLA-A*24:02 were established to determine epitope peptide by screening A*24:02-restricted TCR. The screened TCRs were replaced with mouse C regions and packaged with lentiviruses. Functional validation was performed on healthy human CD4 +T and CD8 +T lymphocytes following infection. Results:Stable TCR-expressing cell lines were successfully prepared based on single-cell TCRαβ double-chain amplification and pairing technology. Twenty-one TCRs were screened using immortalized B lymphocytes, resulting in nine possible HLA-A*24:02-restricted HBsAg-specific TCRs. Further screening with K562-A2402 resulted in six A*24:02-restricted HBsAg-specific TCRs with identically recognized epitope peptide. The functional determination of the two TCR clones revealed their specific recognition function for target cells expressing HBsAg.Conclusion:HLA-A*24:02-restricted HBsAg-specific TCR with recognition function for target cells expressing HBsAg was successfully obtained based on the new experimental technology system, laying an important foundation for further exploration of antiviral immunotherapy based on HBV-specific TCR.

Objective:To investigate the long-term characteristic changes of virus, immune status, and liver fibrosis markers in human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infected patients after receiving direct-antiviral agents (DAAs).Methods:HIV/HCV co-infected patients who visited the Guangzhou Eighth People’s Hospital, Guangzhou Medical University from May 2014 to December 2019 were selected as the research subjects. The changes of virological response rate, peripheral blood CD4 +T lymphocyte level and serological markers of liver fibrosis (APRI score and FIB-4 index) were observed during 144 weeks of follow-up course after the end of DAAs treatment. Kruskal-Wallis test was used for statistical approach. Results:A total of 103 cases were included in the study. There were 87 males (87.5%), with a median age of 44 years. Sustained virological response rate at 12 weeks (SVR12) after DAAs treatment was 97.6%, and the SVR during the entire follow-up period was at least 95.9%. Compared with baseline, CD4 +T lymphocyte count were significantly increased equally at 12 weeks ( Z = -2.283, P = 0.022), 24 weeks (Z = -3.538, P < 0.001), 48 weeks ( Z = -3.297, P = 0.001), 96 weeks ( Z = -3.562, P < 0.001), and 144 weeks ( Z = -2.842, P = 0.004). APRI score ( Z = -6.394, P < 0.001) and FIB-4 index ( Z = -2.528, P = 0.011) were significantly lower than baseline at week 4 of DAAs treatment, and thereafter remained at a low level, without further declination. Conclusion:HIV/HCV co-infected patients can maintain high SVR for a long time, acquire good immune reconstitution, and significantly improve liver fibrosis after DAAs treatment.

Objective:To investigate the immunological features of 135 patients with corona virus disease 2019 (COVID-19), and to provide reference for the pathogenesis of the disease.Methods:The clinical and laboratory data of 135 confirmed COVID-19 patients in Guangzhou Eighth People′s Hospital from January 23 to February 29, 2020 were collected. The features of lymphocytes (CD4 + and CD8 + T lymphocytes, B lymphocytes, natural killer cells and natural killer T cells), and cytokines (interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α and interferon (IFN)-γ) of patients at a median of 19 (14, 27) days of admission were analyzed. Kruskal-Wallis test, Mann-Whitney U test, chi-square test and Spearman rank correlation were used for statistical analysis. Results:Patients were divided into three groups according to the relevant diagnostic criteria, including mild group (14 cases), ordinary group (92 cases) and severe group (29 cases). Decreased CD4 + T lymphocytes were found in 44.4% (60/135) patients, while decreased CD8 + T lymphocytes were found in 42.2%(57/135) patients. Compared to mild group and ordinary group, level of CD4 + T lymphocytes in severe group was significant lower ( Z=4.379 and 3.799, respectively, both P<0.01), and level of CD8 + T lymphocytes was also significant lower ( Z=2.684 and 3.306, respectively, P=0.022 and 0.003, respectively). Decreased B lymphocytes were found in 25.3% (24/95) patients and significant different among the three groups, with the lowest levels ((88(56, 189)/μL; Z=6.199, P=0.045) and most frequency of decreased levels ((52.2%(12/23); χ2=11.723, P=0.003) in the severe group. Compared to the mild group and the ordinary group, IL-6 level in severe group was significant higher ( Z=-4.022 and -4.108, respectively, both P<0.01) and IL-10 level was also significant higher ( Z=-3.261 and -4.006, respectively, both P<0.01). Similar levels of IL-2, IL-4, TNF-α and IFN-γ were found among three groups (all P>0.05). The IL-6 level was positively correlated with the persistence of viral shedding ( r=0.301, P=0.007). Conclusion:The immune-mediated inflammation may be the important cause of disease deterioration of COVID-19, which might be the key target of the treatment of severe cases.

Objective: To analyze the characteristic mutations of epitopes in HBV Pre-S/S region in HIV/HBV co-infected patients’ peripheral blood to provide basic data for studying the pathogenesis of HIV/HBV co-infection. Methods: The chronic hepatitis B infected patients admitted to the Infectious Disease Center of the Eighth People′s Hospital of Guangzhou from January 2009 to December 2011 were enrolled into HIV/HBV co-infected group and HBV mono-infected group according to the result of HIV antibody detection respectively before treatment. HBV DNA in serum was extracted and Pre-S/S region of HBV DNA was amplified by nested-PCR. After sequencing of the obtained PCR products (direct sequencing), ContigExpress software was used for sequence splicing and BioEdit software was used for sequence alignment. With reference to the standard sequence of the matched genotype HBV, mutants of HBV Pre-S/S region in HIV/HBV co-infected group and HBV mono-infected group were analyzed respectively. Statistical analysis was performed by chi-square test with SPSS19.0 statistical analysis software. Results: HBV Pre-S/S fragments were successfully amplified from 150 patients, including 90 cases of HIV/HBV co-infected group and 60 cases of HBV mono-infected group, with matched gender, age, genotype, HBeAg status, alanine aminotransferase (ALT), aspartate aminotransferase (AST). The result of analyzing mutants of HBV Pre-S/S region indicated that the incidence of mutation in all epitopes for cytotoxic T cells (CTL cells) was higher in the HIV/HBV co-infected group, and Pre-S2 aa1-15 epitope was significantly higher (χ²=6.964, P=0.008). The incidence of deletions in PreS2 aa1-15 epitope in HIV/HBV co-infected group (11.1%) was higher than HBV mono-infected group (3.3%) (χ²=2.959, P=0.085). In the B cell epitopes, the incidence of mutations in Pre-S2 aa1-26 in the HIV/HBV co-infected group was significantly higher than HBV mono-infected group (χ²=6.924, P=0.010), and there was no statistical significance between two groups in other B cell epitopes. No differences in helper T cell (Th cell) epitopes were found between the two groups. Conclusions Co-infection with HIV increased the CTL cell epitopes’ mutations in the HBV Pre-S/S region, especially the 5′ end epitope mutations in Pre-S2 region, which indicated that HBV mutation is related to the host immune status, and showed guiding information for further study on the pathogenesis of HIV/HBV co-infection

Objective: To investigate the clinical, immunological and virological characteristics of HIV-1 infected patients in the acute phase, for the sake of improving the diagnosis of acute infection with HIV-1. Methods: We retrospectively analyzed the clinical manifestation and laboratory data of patients with acute HIV-1 infection who were admitted to the Center of Infectious Diseases, Guangzhou Eighth People’s Hospital from January 2012 to June 2017. Results: Forty-four patients were enrolled into the study, 86.4% of them were male. 59.1% patients were homosexually transmitted. Clinical symptoms and signs mostly consisted of fever (84.1%), lymphadenopathy (56.8%) and so on, while 15.9% patients had central nervous system symptoms. Most common opportunistic infection included lung infection (50.0%) and oropharyngeal candidiasis (22.7%). Leucopenia (10 patients, 22.7%), and decreased CD4⁺ T cell count (267.5 cells/μl), inverted CD4⁺ /CD8⁺ ratio (86.4%) was mostly seen. Compared to patients who had HIV RNA load less than 6 lg copies/ml, the group of patients who had HIV RNA load more than 6 lg copies/ml had lower levels of CD4⁺ T cells (t=-3.724, P=0.001). Conclusions Patients with acute HIV infection have many different kinds of clinical symptoms and can be accompanied by opportunistic infection, and with high viremia.