Alcoholic liver disease (ALD) is a growing global health concern, and its early pathogenesis includes steatosis and steatohepatitis. Inhibiting lipid accumulation and inflammation is a crucial step in relieving ALD. Evidence shows that puerarin (Pue), an isoflavone isolated from Pueraria lobata, exerts cardio-protective, neuroprotective, anti-inflammatory, antioxidant activities. However, the therapeutic potential of Pue on ALD remains unknown. In the study, both the NIAAA model and ethanol (EtOH)-induced AML-12 cell were used to explore the protective effect of Pue on alcoholic liver injury in vivo and in vitro and related mechanism. The results showed that Pue (100 mg·kg^-1) attenuated EtOH-induced liver injury and inhibited the levels of SREBP-1c, TNF-α, IL-6 and IL-1β, compared with silymarin (Sil, 100 mg·kg^-1). In vitro results were consistent within vivo results. Mechanistically, Pue might suppress liver lipid accumulation and inflammation by regulating MMP8. In conclusion, Pue might be a promising clinical candidate for ALD treatment.

Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

Objective:To explore the relationship between the changes of anorectal angle (ARA) under 3 physiological states and pelvic organ prolapse(POP) in postpartum women by transperineal ultrasound.Methods:The retrospective study enrolled 147 female in 6-8 weeks after delivery examined by pelvic floor ultrasound examinations in Fujian Medical University Second Affiliated Hospital from November 2019 to June 2021, who were divided into POP group and control group. Volume data of pelvic floor ultrasound examinations were obtained at rest, during contraction and during maximal Valsalva maneuver. The differences in the changes of ARA under 3 physiological states between the two groups were compared, and the correlation between the change state of ARA during maximal Valsalva maneuver and POP was analyzed.Results:Compared with ARA at rest, ARA decreased during contraction (χ 2=42.64, P<0.001) and increased during maximal Valsalva maneuver (χ 2=38.43, P<0.001). There was no difference of ARA between the POP group and control group in the 3 physiological states ( P>0.05). However, the risk of POP increased when ARA decreased during maximal Valsalva maneuver ( OR=2.690, 95% CI=1.074-6.739, P<0.05). Conclusions:The decrease of ARA during maximal Valsalva maneuver may increase the risk of POP, and the change of ARA during maximal Valsalva maneuver can be brought into the ultrasonic observation indicators of POP.

Objective:To assess the interobserver and inter-modalities agreement with two non-invasive diagnostic modalities of hepatocellular carcinoma in high-risk patients: contrast-enhanced ultrasound liver imaging reporting and data system (CEUS LI-RADS) and magnetic resonance imaging liver imaging reporting and data system (MRI LI-RADS).Methods:From August 2017 to August 2019, the CEUS and MRI data of patients at high risk for HCC from the Second Affiliated Hospital of Zhejiang University School of Medicine were analyzed retrospectively. A total of 217 lesions in 173 patients were classified according to CEUS LI-RADS v. 2017 or MRI LI-RADS v. 2018, by 4 blinded independent observers with more than 10 years of experience of CEUS or MRI. Interobserver and inter-modalities agreement was assessed with Cohen′s kappa.Results:The interobserver agreement was moderate and comparable for CEUS/MRI LI-RADS category (κ=0.606/0.603), the inter-modalities agreement was moderate for CEUS and MRI LI-RADS category (κ=0.564), LI-RADS 3, M, 4 and 5 by two imaging methods showed that the Kappa values were 0.739, 0.551, 0.734 and 0.592, respectively.Conclusions:The total inter-modalities agreement between CEUS and MRI LI-RADS categories is moderate, while the agreements of LI-RADS 3, 4 are strong, and LI-RADS M, 5 are moderate.

Objective To investigate the clinical effect of minimally invasive extraction of anterior tooth residual root after root separation.Methods A total of 400 patients receivinganterior tooth residual root extraction were collected in the clinic of oral and maxillofacial surgery department between January 2015 and December 2016.The patients were divided into a control group and a study group according to their sequence to see the doctor,with an odd for the study group and an even for the control group.In the study group,residual roots were separated mesiodistally by high speed turbine before using minimally invasive extraction tool;while in the control group residual roots were extracted only using minimally invasive extraction tool.The surgical duration,postoperative damage rate of the lip side plate,degree of pain and patient satisfaction in the two groups were analyzed.Results The surgical duration was shorter in the study group compared with the control group (P ＜ 0.05).The postoperative damage rate of the lip side plate and the degree of pain were lower,while patient satisfaction was higher in the study group than in the control group (P ＜ 0.05).Conclusions The postoperative damage rate of the lip side plate is significantly lower in minimally invasive extraction of anterior tooth residual root after root separation.Smaller trauma is conducive to the implant afterwards.Root separation in minimally invasive extraction of anterior tooth residual root is valuable for clinical application.

Objective To explore the validity of the size-specific dose estimate (SSDE) derived from the water-equivalent diameter (Dw)value of the slice located in the middle of the scan range in the head CT examination. Methods A total of 197 patients underwent head CT nonenhanced scan were enrolled in this retrospective study. The Dw, size-dependent conversion factor (f), normalized volume CT dose index (CTDIvol) and SSDE values of all slices were calculated. Two sets of SSDE, SSDEgroand SSDEcenbased on the Dwvalues slice by slice (Dw-gro) and the Dwvalues of the slices in the middle of the scan range (Dw-cen), were obtained across all patients. Pearson correlation analysis and linear regression analysis were performed for Dw-grovs Dw-cen, Spearman correlation analysis and linear regression analysis for SSDEgrovs SSDEcen, SSDE vs Dw, CTDIvolvs Dw. With the reference of SSDEgrovalue, mean absolute relative difference (MARD) of SSDEcen values were calculated to assess its accuracy and the correlated factors of MARD was analyzed with multivariate linear stepwise regression analysis. Results The minimal Dwvalue close to the roof of the skull corresponded to the maximal value of f and SSDE, which was the minimal value of CTDIvol. The significant positive correlation was showed between Dw-grovs Dw-cen, SSDEgrovs SSDEcen, SSDE vs Dw, CTDIvolvs Dw(r=0.947, 0.931, 0.416, 0.626;P＜0.05). The values of Dw,groand Dw-cenwere (16.94±0.69) and (18.50±0.62) cm respectively. The values of SSDEgroand SSDEcenwere [54.10 (52.29, 56.39)] mGy and [53.77 (51.85, 55.25)] mGy respectively. An approximation of SSDEcenvalues with an average of 1.62% of the gross MARD was found to match the reference value. Multivariate linear stepwise regression analysis indicated that MARD had negative correlation with Dw(β=–1.319,P＜0.05), positive correlation with CTDIvol(β=0.202,P＜0.05), and f was not included in the multivariate regression equation. Conclusion SSDEcenbased on the Dwvalue of the slice located at the center of the scan range yields small MARD value and can represent a reliable SSDE estimation in the head CT examination.

Objective To investigate the effects of epidural ropivacaine block combined with propofol intravenous anesthesia on CaMKⅡ and ERK1/2 total protein (T-CaMKⅡ and T-ERK1/2) and phosphorylation(p-CaMKⅡ and p-ERK1/2) levels in the hippocampus and cortex of rats.Methods Rats were randomly assigned to three groups: group P(control,propofol intravenous anesthesia),group PS(propofol and epidural normal saline) and group PR(propofol and epidural 0.5% ropivacaine).Anesthesia were performed in 72 h after epidural catheter placement.The rats in group PR received 70 μL of 0.5% ropivacaine to achieve epidural block.1% propofol was infused through rats caudal vein.Propofol dosage for anesthesia induction was 12 mg/kg,for anesthesia maintenance was 40 mg/(kg·h).Before the rats were decapitated,the depth of anaesthesia was assessed as either light anesthesia or deep anesthesia by checking of pinch withdrawalreflex,eyelid reflex and spontaneous rapid whisking of the vibrissae after propofol continuous infusion for 1 h.T-CaMKⅡ/T-ERK1/2 and p-CaMKⅡ/p-ERK1/2 in hippocampus and frontal cortex were examined by Western blot.Results 7 rats were assessed as light anesthesia and one rat as deep anesthesia in group P;6 rats were assessed as light anesthesia and 2 rats as deep anesthesia in group PS;in group PR,1 rat was assessed as light anesthesia and 7 rats as deep anesthesia.Significant differences were seen among three groups (P<0.05).In hippocampus of rats,p-CaMKⅡ(Thr286)43.7%±8.8% and p-ERK1/2 32.4%±7.9% in group PR were significantly lower than those in group P (100%,P<0.05).Conclusions Epidural ropivacaine block may strengthen the depth of anesthesia achieved with propofol intravenous anesthesia.The decrease of p-CaMKⅡ(Thr286) and p-ERK1/2 in hippocampus of rats may explain the effects of epidural block.

We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.

Objective:To establish a double antibody sandwich ELISA assay for detection of human IL-37 in serum.Methods: Mouse anti-human IL-37 monoclonal antibody was used as capturing antibody,rabbit anti-human IL-37 polyclonal antibodies served as detection antibody,HRP labeled goat anti-rabbit IgG employed as second antibody and recombinant human IL-37 protein used as reference standard for the establishment of a doubleantibody sandwich ELISA.The working conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients with Dengue fever.Results: The sensitivity of the established ELISA method was 1.465 μg/L approximately.Likewise,the linearity range of this method was about (1.465-46.875) μg/L.Further,the co efficient of variation (CV) of inter-batch and intra-batch in this study were 6.6％ and 11.7％,respectively.Notably,this method could be used in the detection of IL-37 in serum of the patients with Dengue fever,showing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls.Conclusion: The double antibody sandwich ELISA assay for the detection of human IL-37 was successfully established,which can be apply to detect of human IL-37 in clinical samples.

Objective To investigate the level change of interleukin-37(IL-37) and its clinical significance in the patients with chronic kidney disease(CKD) and nephrotic syndrome(NS) .Methods 57 cases of CKD (CKD group) and 13 cases of NS (NS group) in the Affiliated Donghua Hospital of Sun Yat-sen University from September to November 2016 were selected .The CKD group was redivided into the CKD stage 1 ,stage 3 and stage 5 groups .At the same time 22 individuals undergoing physical examina-tion were selected as the healthy control group .The ELISA sandwich method was adopted to detect the plasma IL-37 level .Then the detection results were statistically analyzed .Results The level of plasma IL-37 in the CKD group and NS group was significant-ly higher than that in the healthy control group ,the difference was statistically significant (P<0 .01);the level of plasma IL-37 had no statistical difference among the CKD stage 1 group ,CKD stage 3 group and CKD stage 5 group(P>0 .05);the plasma IL-37 lev-el after treatment in the CKD group and NS group was decreased compared with before treatment ,the difference was statistically significant(P<0 .01);the plasma IL-37 level in CKD patients was positively correlated with the WBC and lymphocyte cells levels . Conclusion The patients with CKD and NS have high level expression of plasma IL-37 ,which might have good auxiliary diagnostic value for CKD and NS .