Objective:To analyze the mutation of T-cell and B-cell epitopes derived from HBV PreS-S protein in occult hepatitis B virus (OHBV) and investigate the biological mechanisms of occult hepatitis B virus infection (OBI) and HBsAb positive OBI.Methods:The PreS-S region of OBI samples were amplified by nested PCR, the products were sequenced and HBV genotypes were determined. The mutations of T-cell and B-cell epitopes derived from HBV PreS-S protein were analyzed and compared among groups of HBV genotypes and the presence of HBsAb. The affinity of the high frequency of T-cell epitope substitutions were analyzed by SYF PEITHI, the changes of antigenic characteristics of high frequency of B-cell epitope substitutions were analyzed by Ab Designer, Expasy ProtParam tool, Epitope Prediction and Analysis Tools.Results:The PreS-S region of HBV was amplified in 21 samples, including 4 HBsAb+ OBI B, 6 HBsAb-OBI B, 6 HBsAb+ OBI C, 5 HBsAb-OBI C. The mutation rates in PreS-S region of OBI were significantly higher than wild type HBV strains(OBI Bvs. WT B: 2.64%: 0.66%, P<0.001; OBI Cvs. WT C: 3.67%: 1.19%, P<0.001). The mutation rates of the immunoreactive area were significantly higher than non-immunoreactive area in OBI (OBI B: 3.57%: 1.86%, P=0.005; OBI C: 4.78%: 2.65%, P<0.001). The mutation rates of the immunoreactive and non-immunoreactive area in OBI C were higher than OBI B, but there was no statistically significant difference (immunoreactive area: 4.78%: 3.57%, P=0.107; non-immunoreactive area: 2.65%: 1.86%, P=0.142). The mutation rates of T-cell and B-cell epitopes of HBsAb-OBI were higher than HBsAb+ OBI, although there was no significant difference (HBsAb-OBI Bvs. HBsAb+ OBI B: 4.17∶3.01, P=0.303; HBsAb-OBI Cvs. HBsAb+ OBI C: 5.65∶4.26, P=0.207). The affinity analysis of 4 high frequency T-cell epitope substitutions, including T47A/K, S174N, L175S, V177A, showed that the changes of affinity of most mutation sites were not obvious; the antigenicity analysis of 3 high frequency B-cell epitope substitutions, including G73S, K122R, I126M/T, did not show noticeable changes and the hydrophilicity, surface accessibility of some mutation sites were even better than wild strain. Conclusions:The mutation rates in PreS-S region of OBI were significantly higher than wild type HBV strains. The mutation rates of the immunoreactive area were higher than non-immunoreactive area in OBI. The variant activity of OBI C was higher than OBI B. The mutations of OBI might occur randomly and were not selected by antibody pressure. Single epitope and multi-epitopes combinational mutations might be a reason for OBI.

Objective: Poor experience with Invisalign treatment affects patient compliance and, thus, treatment outcome. Knowing the potential discomfort level in advance can help orthodontists better prepare the patient to overcome the difficult stage. This study aimed to construct artificial neural networks (ANNs) to predict patient experience in the early stages of Invisalign treatment. Methods: In total, 196 patients were enrolled. Data collection included questionnaires on pain, anxiety, and quality of life (QoL). A four-layer fully connected multilayer perception with three backpropagations was constructed to predict patient experience of the treatment. The input data comprised 17 clinical features. The partial derivative method was used to calculate the relative contributions of each input in the ANNs. Results: The predictive success rates for pain, anxiety, and QoL were 87.7%, 93.4%, and 92.4%, respectively. ANNs for predicting pain, anxiety, and QoL yielded areas under the curve of 0.963, 0.992, and 0.982, respectively. The number of teeth with lingual attachments was the most important factor affecting the outcome of negative experience, followed by the number of lingual buttons and upper incisors with attachments. Conclusions The constructed ANNs in this preliminary study show good accuracy in predicting patient experience (i.e., pain, anxiety, and QoL) of Invisalign treatment. Artificial intelligence system developed for predicting patient comfort has potential for clinical application to enhance patient compliance.

[ABSTRACT]AIM:ToobservehowfarnesoidXreceptor(FXR)functionedinconcanavalinA(ConA)-induced hepatitis (CIH) and the regulation of FXR-thyrotropin embryonic factor (TEF) pathway.METHODS:C57BL/6 mice were injected with Con A to induce hepatitis .The expression of FXR and TEF in the liver specimens was determined by qRT-PCR and Western blotting .The concentrations of serum ALT/AST and inflammatory cytokines IFN-γ, TNF-α, IL-4 and IL-2 in the blood samples were tested after Con A injection .RESULTS:FXR was down-regulated in CIH mice .TEF was up-regula-ted when FXR was activated by chenodeoxycholic acid (CDCA).Activation of FXR reduced the levels of aminotransferases and inflammatory cytokines IFN-γ, TNF-α, IL-4 and IL-2 in the CIH mice induced by Con A injection .CONCLUSION:FXR activation attenuates CIH mouse liver injury and reduces inflammatory cytokines .FXR activation results in TEF up-regu-lation.The FXR-TEF pathway may play a protective role in autoimmune hepatitis .

OBJECTIVETo determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).

METHODSPeripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot.

RESULTSLevels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA.

CONCLUSIONIL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.

Adolescent ; Adult ; Cells, Cultured ; DNA-Binding Proteins ; metabolism ; Humans ; Interleukin-12 ; blood ; genetics ; Leukocytes, Mononuclear ; metabolism ; Lupus Erythematosus, Systemic ; metabolism ; Middle Aged ; Phosphorylation ; RNA, Messenger ; analysis ; STAT3 Transcription Factor ; STAT4 Transcription Factor ; Trans-Activators ; metabolism

AIM:To investigate the regulatory role of nuclear factor κB (NF-κB) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1β.METHODS:Activation of NF-κB was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS:rhIL-1β could rapidly stimulate the activation of NF-κB in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-κB， could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1β.CONCLUSION:IL-6 expression induced by IL-1β may be regulated by NF-κB in MC, NF-κB may modulate the immune-inflammatory reaction in glomerular disease.

【Objective】To observe the expression of PI3-K phosphorylated products and elucidate the correlation between PI3-K phosphorylated products and Th2 cytokine in peripheral blood mononuclear cell (PBMC).【Methods】14 patients with active lupus nephritis and 12 controls were selected,PI3-K phosphorylated products were detected by immunoprecipitation and Western blotting,RT-PCR was used to observe interleukin-6 mRNA and interleukin-10 mRNA expression.【Results】In either spontaneous condition or stimulated by anti-CD3 antibody,the expression of PI3-K phosphorylated products in patients with active lupus nephritis were higher than those of the controls(1.14±0.23 vs 0.46±(0.12，P=0.023;2.09±0.63 vs 0.65±0.14，P=0.016).The expression of PI3-K phosphorylated products in active lupus nephritis showed a positive correlation with interleukin-6 mRNA and interleukin-10 mRNA (r=0.652,P=0.008;r=0.718,P=0.007).PY294002,one of specific inhibitor of PI3-K,inhibited significantly the expression of interleukin-6 mRNA(2.32±0.51 vs 0.57±0.15,P=0.009) and interleukin-10 mRNA (1.71±0.33 vs 0.67±0.11,P=0.006) in stimulated PBMC in active lupus nephritis.【Conclusion】PI3-K can involve in the pathogenesis of lupus nephritis by inducing the overexpression of interleukin-6 and interleukin-10.

AIM: To examine whether Akt signal pathway proteins, including Akt, NF-?B and I?B?, are activated in kidney tissue of murine chronic graft-versus -host disease (GvHD) lupus nephritis in vivo , and whether prednisone suppres ses activation of them. METHODS: Akt activity and phosphorylated I?B? were detected by Weste rn-blot. Activation of NF-?B was detected by electropheretic mobilit y shift assay (EMSA). RESULTS: Activity of Akt, NF-?B and phosp horylated I?B ? were significantly increased in kidney tissue of murine chronic graft-versus -ho st disease (GvHD) in 8th week and 12th week after monocell injection, respective ly. However, they were no significant elevation in 16th week, when compared with controls. Prednisone treatment significantly prevented the increase in serum an ti-dsDNA antibody level, urinary protein excretion and glomerular cell prolif eration in GvHD mice, indicating the beneficial effects of prednisone on t his model. Prednisone also significantly suppressed the increase in the activities o f glomerular Akt, NF-?B and phosphorylated I?B?. CONCLUSION: T his study provides t he first evidence of marked increase in glomerular Akt-NF-?B signal pathway act ivities in murine chronic graft-versus-host disease lupus nephritis. The benefic ial effect of prednisone on this lupus nephritis model may be partially mediated by the suppression of abnormal Akt- NF-?B activation.

AIM:To observe the expression of chemokine fractalkine,and its receptor,CX3CR1,in kidneys of lupus-prone BXSB mice,and their changes after treatment with prednisone. The role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was also discussed. METHODS:Twelve 12-week-old male BXSB mice were randomly divided into two groups,the prednisone treatment group (BXSB-prednisone group,n=6) and the experimental control group (BXSB group,n=6). Six male C57BL/6J mice at the same weeks of age served as a normal control group (C57BL/6J group). Both the C57BL/6J and the BXSB group of mice received a daily intragastric administration of 0.5 mL normal saline. The BXSB-prednisone group of mice was given a daily intragastric administration of prednisone (0.18 mg/20 g BW) dissolved in 0.5 mL normal saline. All treatments lasted for 10 weeks. The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of mice were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis respectively. The changes of laboratory index and the kidney histopathology of mice were also investigated. RESULTS:The mRNA and protein expressions of fractalkine and CX3CR1 in kidneys of BXSB mice were significantly higher than those in C57BL/6J mice. The expressions of fractalkine and CX3CR1 in BXSB-prednisone group of mice were much lower than those in BXSB group of mice,accompanied by the lower serum IgG,IgM and anti-dsDNA antibody levels as well as blood urea nitrogen,serum creatinine and urine protein. The glomerular immune complex deposition and the kidney histopathology were also significantly improved in BXSB-prednisone group of mice. CONCLUSION:These results indicate that fractalkine and CX3CR1 participate in the pathogenesis of lupus nephritis in BXSB mice,and the effect of glucocorticoids treatment may be attributed,in part,to its ability to inhibit the expression of fractalkine in kidney.

AIM: To investigate the regulatory role of nuclear factor ?B (NF-?B) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1?.METHODS: Activation of NF-?B was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS: rhIL-1? could rapidly stimulate the activation of NF-?B in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-?B, could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1?.CONCLUSION: IL-6 expression induced by IL-1? may be regulated by NF-?B in MC, NF-?B may modulate the immune-inflammatory reaction in glomerular disease.

AIM: To detect the association between the polymorphism of Fc receptor ? chain gene at position-29 in promoter and systemic lupus erythematosus(SLE). METHODS: The genotypes at position -29 in promoter of Fc receptor ? chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China. RESULTS: The frequencies of TT genotype(33.3%) and T allele (54 4%) at position -29 in patients with SLE were significantly higher than those in controls (17 9%, respectively), whereas, the frequencies of GG genotype (24 4%) and G allele (45 6%) in patients with SLE were remarkably lower than those in controls (31 4% and 57 1%, respectively) ( P 0 05) . CONCLUSION: Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.