Systemic antibiotics increase microbiota pathogenicity and oral bone loss / 国际口腔科学杂志·英文版

Periodontitis is the most widespread oral disease and is closely related to the oral microbiota. The oral microbiota is adversely affected by some pharmacologic treatments. Systemic antibiotics are widely used for infectious diseases but can lead to gut dysbiosis, causing negative effects on the human body. Whether systemic antibiotic-induced gut dysbiosis can affect the oral microbiota or even periodontitis has not yet been addressed. In this research, mice were exposed to drinking water containing a cocktail of four antibiotics to explore how systemic antibiotics affect microbiota pathogenicity and oral bone loss. The results demonstrated, for the first time, that gut dysbiosis caused by long-term use of antibiotics can disturb the oral microbiota and aggravate periodontitis. Moreover, the expression of cytokines related to Th17 was increased while transcription factors and cytokines related to Treg were decreased in the periodontal tissue. Fecal microbiota transplantation with normal mice feces restored the gut microbiota and barrier, decreased the pathogenicity of the oral microbiota, reversed the Th17/Treg imbalance in periodontal tissue, and alleviated alveolar bone loss. This study highlights the potential adverse effects of long-term systemic antibiotics-induced gut dysbiosis on the oral microbiota and periodontitis. A Th17/Treg imbalance might be related to this relationship. Importantly, these results reveal that the periodontal condition of patients should be assessed regularly when using systemic antibiotics in clinical practice.

Four-Octyl itaconate ameliorates periodontal destruction via Nrf2-dependent antioxidant system / 国际口腔科学杂志·英文版

Periodontitis is a widespread oral disease characterized by continuous inflammation of the periodontal tissue and an irreversible alveolar bone loss, which eventually leads to tooth loss. Four-octyl itaconate (4-OI) is a cell-permeable itaconate derivative and has been recognized as a promising therapeutic target for the treatment of inflammatory diseases. Here, we explored, for the first time, the protective effect of 4-OI on inhibiting periodontal destruction, ameliorating local inflammation, and the underlying mechanism in periodontitis. Here we showed that 4-OI treatment ameliorates inflammation induced by lipopolysaccharide in the periodontal microenvironment. 4-OI can also significantly alleviate inflammation and alveolar bone loss via Nrf2 activation as observed on samples from experimental periodontitis in the C57BL/6 mice. This was further confirmed as silencing Nrf2 blocked the antioxidant effect of 4-OI by downregulating the expression of downstream antioxidant enzymes. Additionally, molecular docking simulation indicated the possible mechanism under Nrf2 activation. Also, in Nrf2-/- mice, 4-OI treatment did not protect against alveolar bone dysfunction due to induced periodontitis, which underlined the importance of the Nrf2 in 4-OI mediated periodontitis treatment. Our results indicated that 4-OI attenuates inflammation and oxidative stress via disassociation of KEAP1-Nrf2 and activation of Nrf2 signaling cascade. Taken together, local administration of 4-OI offers clinical potential to inhibit periodontal destruction, ameliorate local inflammation for more predictable periodontitis.

Effect of artificial liver support therapy on the short-term prognosis of patients with liver failure in the plateau stage: A stratified analysis based on Model for End-Stage Liver Disease score / 临床肝胆病杂志

ObjectiveTo investigate the effect of artificial liver support therapy on the short-term (28- and 90-day) mortality rate of patients with liver failure in the plateau stage through a stratified analysis based on Model for End-Stage Liver Disease (MELD) score. MethodsA retrospective analysis was performed for 187 patients with liver failure who were admitted to Nanfang Hospital, Southern Medical University, from January 2015 to April 2019, with 73 patients in the artificial liver group and 114 in the non-artificial liver group. The stratified analysis based on MELD score in the plateau stage was performed to investigate the differences in 28- and 90-day mortality rates, hospital costs and length of hospital stay of surviving patients, and incidence rate of adverse reactions of artificial liver support therapy between the two groups. The t-test was used for comparison of continuous data between the two groups, and the chi-square test or the Fisher’s exact test was used for comparison of categorical data between the two groups. ResultsCompared with the non-artificial liver group, the artificial liver group had a significant reduction in the 28-day mortality rate of the patients with an MELD score of 30-39 (5.9% vs 39.6%, P＜0.001) or those with an MELD score of 40 (25.0% vs 72.7%, P＜0.05). Compared with the non-artificial liver group, the artificial liver group had a significant reduction in the 90-day mortality rate of the patients with an MELD score of 30-39 (23.5% vs 623%, P＜0.001). Artificial liver support therapy did not significantly shorten the mean hospital stay of the surviving patients (P＞0.05) and had no significant influence on the total hospital costs of the surviving patients within 90 days (P＞0.05). The incidence rate of adverse reactions related to artificial liver support therapy was 29.1%, but the symptoms were mild and were relieved after symptomatic treatment. ConclusionPatients with an MELD score of ＜30 in the plateau stage tend to have low 28- and 90-day mortality rates, and artificial liver support therapy can be reasonably selected according to the patient’s economic conditions and willingness. Artificial liver support therapy is recommended for patients with an MELD score of 30-39 in the plateau stage if there is no obvious contraindication. For patients with an MELD score of 40 in the plateau stage, artificial liver support therapy is recommended within 28 days if there is no obvious contraindication, and liver transplantation is recommended as soon as possible. Artificial liver support therapy has no significant influence on the total hospital costs and mean hospital stay of the surviving patients within 90 days and does not increase the economic burden of patients.

Association of baseline alanine aminotransferase levels with therapeutic effects of entecavir and interferon- in patients with chronic hepatitis B / 南方医科大学学报

OBJECTIVE@#To evaluate the therapeutic effects of entecavir (ETV) and interferon- (IFN-) treatments for 48 weeks for chronic hepatitis B (CHB) in patients with different baseline alanine aminotransferase (ALT) levels.@*METHODS@#We retrospectively analyzed the data of 369 CHB patients receiving ETV and IFN- treatments for 48 weeks. We compared the virological response rates, HBsAg clearance, and HBsAg reduction between the patients receiving ETV and IFN- treatments with different baseline ALT levels[≤ 5×upper limits of normal (ULN) level (subgroup 1), 5-10×ULN (subgroup 2), and > 10× ULN (subgroup 3)].@*RESULTS@#In patients receiving ETV treatment, the virological response rate was 83.3% in subgroup 1, 91.4% in subgroup 2, and 95.5% in subgroup 3, as compared with 19.7%, 40%, and 42.9% in the 3 subgroups with IFN- treatment, respectively, showing significantly differences both among different subgroups with the same treatment and between the same subgroup with different treatments ( < 0.05). HBeAg clearance rates in the 3 subgroups were 8.3%, 16.7% and 35.5% in patients with ETV treatment and were 1.8%, 41.9%, and 38.1% in patients with IFN- treatment, respectively, showing significant differences among the 3 subgroups with the same treatment ( < 0.05); in the same subgroups with different treatments, the rates differed significantly only between subgroups 2 ( < 0.05). In ETV group, the rate of HBsAg reduction to below 200 IU/ml was 2.5% in subgroup 1 and 13.8% in subgroup 2, showing no significant difference between the two subgroups; in IFN- group, the rates were also similar between subgroups 1 and 2 (30.6% 33.3%, > 0.05); but the rates differed significantly between the same subgroups with different treatments ( < 0.05).@*CONCLUSIONS@#In all the subgroups with different baseline ALT levels, ETV treatment for 48 weeks results in significantly higher virological response rates than IFN- treatment in patients with CHB. In patients with a baseline ALT of 5-10 ×ULN, IFN- can result in a higher HBeAg clearance rate than ETV. In patients with comparable baseline ALT level, IFN- more effectively reduces HBsAg level than ETV. The patients with a relatively high baseline ALT level (> 5 × ULN) show better responses to both ETV and IFN- treatment than those with ALT level below 5×ULN. We thus recommend IFN- for patients with a baseline ALT of 5-10×ULN and ETV for patients with a baseline ALT either below 5 × ULN or beyond 10×ULN.

Association between APOBEC3G polymorphisms and susceptibility to chronic hepatitis B / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association between rs185983011 single-nucleotide polymorphisms (SNP) of apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G (APOBEC3G) and the susceptibility to chronic hepatitis B.</p><p><b>METHODS</b>The blood samples were collected from 186 healthy subjects and 159 patients with chronic hepatitis B. The rs185983011 SNP was detected and genotyped by sequencing with Sanger's method to analyze the relationship between rs185983011 SNP and chronic hepatitis B.</p><p><b>RESULTS</b>Only C/C and C/T genotypes of the alleles of rs185983011 SNP were found in the tested subjects, and the C/C genotype was predominant (97.7%). The distribution frequencies of rs185983011 SNP genotypes and alleles showed no significant difference between healthy subjects and patients with chronic hepatitis B (P>0.05).</p><p><b>CONCLUSION</b>The predominant genotype of rs185983011 SNP of APOBEC3G is C/C in the tested subjects, and rs185983011 SNP does not appear to associate with the susceptibility to chronic hepatitis B.</p>

Analysis of the factors for predicting the outcomes of interferon-α and entecavir treatments for chronic hepatitis B with positive HBeAg / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the predictive factors of the therapeutic effects of interferons (IFNs) and entecavir (ETV) treatments for 48 weeks in patients with chronic hepatitis B (CHB) positive for HBeAg.</p><p><b>METHODS</b>This retrospective analysis compared the treatment efficacy of IFNs and ETV in 129 CHB patients positive for HBeAg. Twenty-seven of the patients were treated with PEG-IFNα-2a (180 µg once a week, PEG-IFN group), 51 patients with conventional IFNα (5 MIU three times a week, IFN group), and 51 with ETV (0.5 mg once daily, ETV group) for 48 weeks.</p><p><b>RESULTS</b>After completion of the treatment cycles, the patients in ETV group showed a significantly higher HBV DNA undetectable rate and a significantly lower HBeAg seroconversion rate than those in PEG-IFN and IFN groups (P<0.05); HBeAg seroconversion rates were similar between PEG-IFN group and IFN group (Χ(2)=0.709, P=0.400). In PEG-IFN and ETV groups, HBeAg seroconversion rates were not associated with age, gender, baseline HBeAg, baseline HBV DNA and baseline ALT. In IFN group, HBeAg seroconversion rates were associated with baseline HBeAg (P=0.048) but not with age, gender, baseline HBV DNA and baseline ALT. In PEG-IFNα-2a group, ROC analysis showed that the sensitivity and specificity of HBeAg seroconversion at 48 weeks were 0.778 and 0.889, respectively, when the decline rate of HBeAg between baseline and week 24 exceeded 97.81%, with the corresponding positive and negative predictive values (PPV and NPV) of 0.778 and 0.889, respectively; the sensitivity and specificity of HBeAg seroconversion at 48 weeks were 0.889 and 0.722, respectively, when the decline rate of HBeAg between week 12 and week 24 was over 42.75%, with the corresponding PPV and NPV of 0.615 and 0.929, respectively.</p><p><b>CONCLUSION</b>Treatments with PEG-IFNα-2a and conventional IFNα for 48 weeks can achieve a higher HBeAg seroconversion rate than ETV, but the latter produces a higher HBV DNA undetectable rate. For PEG-IFNα-2a treatment, the decline rate of HBeAg between baseline and week 24 over 97.81% is the best predicting factor for HBeAg seroconversion at week 48 in CHB patients positive for HBeAg.</p>

Cloning and expression and purification of hepatitis B e-antigen precursor in Escherichia coli / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of the 25 kD hepatitis B e antigen (HBeAg) precursor that only exist inside hepatocytes and study its effect on the pathopoiesis of hepatitis B and QIAGEN expression and purification system.</p><p><b>METHODS</b>Hepatitis B virus (HBV) preC/C gene for the 25 kD HBeAg precursor was cloned into the expression vector pQE30 and the 25 kD HBeAg precursor was expressed in Escherichia coli (E. coli) and purified. Its antigenicity for 21 kD mature HBeAg was tested by western blot analysis.</p><p><b>RESULTS</b>Cloned fragments in the expression vector were sequenced and verified to be homogeneous with that of HBV (ayw subtype). Expression of the HBeAg precursor in E. coli under the transcriptional regulation of T5 promoter yielded a soluble cytosolic protein with an apparent molecular mass of 25 kD. Recombinant HBeAg precursor exhibited identical potencies with 21 kD mature HBeAg that reacted with anti-HBeAg antibodies. The purification rate of the expressed HBeAg precursor was up to 89.6% and the yield of purified HBeAg precursor from this procedure was 2.4 mg/L.</p><p><b>CONCLUSION</b>25 kD HBeAg precursor exhibited biological activity and might play an important role in pathopoiesis of hepatitis B.</p>

ESTABLISHMENT AND CHARACTERISTICS OF Epstein-Barr VIRUS TRANSFORMED B CELL LINES FROM HEPATITIS B PATIENTS / 解放军医学杂志

To study the characteristics of EBV transformed human peripheral blood B cell lines from hepatitis B patients and to provide a basis for further studies on the cellular immune function of hepatitis B patients.High titer of EBV was produced by B95 8 cell by induction with sodium n butyrate,and peripheral mononuclear cells from two different groups of hepatitis B patients were harvested and infected with EBV. The results showed that 4～8 weeks after infection 19 EBV transformed B cell lines from hepatitis B patients were established. It was found that colony formation of cells from A group appeared 2～3 weeks later than B group. Cells grew healthily and had good activity after 4 weeks of freezing.CD19 and CD20 were detected in 88 34% and 37 48% of the immortalized B cell membrane respectively. HBV DNA could not be detected in a11 19 immortalized B cell lines.It suggested that the time needed for the establishment of B cell lines was related with the immune state of the patients.HBV DNA could not exist persistently, and it would disappear finally in the immortalized B cell lines.

Construction of mutant gene of HBV and investigating its biological significance / 中华传染病杂志

Objective To study the biological significance of the common mutant preC/C gene in clinical HBV in China. Methods Site directed mutagenesis based on the unique enzyme site elimination was used to construct eukaryocyte expression vector with mutant HBV/C gene(V60、G87、L97). Expression vectors with wild and mutant preC/C gene were transferred into HepG2 cell. Culture supernatant was detected by ELISA for HBeAg. Results Result of DNA sequencing showed that the constructed mutant HBV preC/C gene had only one specific site variation compared with the wild type sequence. Goal DNA fragment was detected positive in the HepG2 cells transferred with wild and mutant preC/C gene. A value of HBeAg in the supernatant of the cells harboring L97 variant was higher than that of the wild and other variant strains( P

Construction of hepatitis B virus C gene retroviral vector and its expression in immortalized human peripheral blood B cell lines / 中华传染病杂志

Objective In order to study the biological significance of HBV/C gene variant in vitro.Methods Retroviral vector pXT1 was used to construct the HBV/C gene expression vector and the recombinant plasmid pXT1-HBV/C was used to transfect immortalized human peripheral blood B cell lines to express HBcAg steadily in the host cells.Results plasmid pXT1-HBV/C was detected positive by PCR as well as enzyme digestion with Bgl Ⅱ and Xho Ⅰ.Meanwhile.transfected cells was detected to contain HBV/C gene by PCR and HBcAg was expressed in 47.4%of the ceils by means of flow cytometry.Conclusion Retroviral expression vector with HBV/C gene can transfeet into eucaryotic cells effectively and express the goal gene steadily.The recombinant cells may be used in the systematic studies on the biological significance of HBV/C gene variant.