Background/Aims: Regorafenib is recommended by guidelines and trials as a sequential second-line therapy following progression on first-line sorafenib or lenvatinib in hepatocellular carcinoma (HCC). However, efficacy is limited, highlighting the urgent need to screen suitable patients and develop sensitization strategies. Methods: Acquired sorafenib- or lenvatinib-resistant (SR or LR) HCC cell lines and organoids were established. Genome-wide CRISPR library screen was performed in SR or LR cell strains to identify synthetic lethal targets of regorafenib. RNA-seq and FITC-regorafenib efflux assay were used to elucidate ERBB3-driven downstream signaling. Preclinical mouse models of cell line- and patient-derived xenografts and clinical cohorts of HCC patients were employed to validate the efficacy of ERBB3-guided patient stratification. Results: Screening with CRISPR library, we showed that inhibition of ERBB3 was synthetic lethal with regorafenib in SR or LR cell strains and organoids. Mechanistically, SR or LR triggered feedback activation of ERBB3 signaling and mediated regorafenib efflux via ERBB3-HIF1A-ABCB1 cascade pathway, limiting sensitivity to regorafenib. Moreover, ERBB3-low tumors following SR or LR exhibited significant sensitivity to regorafenib, suggesting its potential as a predictive biomarker to screen optimal candidates for sequential therapy. Seribantumab, an ERBB3-targeting monoclonal antibody, inhibited ERBB3-HIF1A-ABCB1 cascade, and its combination with regorafenib exerted marked synergistic anti-tumor effects on ERBB3-high tumors resistant to sorafenib or lenvatinib both in vitro and in vivo. Conclusions This study revealed that ERBB3 was a key resistance factor driving limited efficacy to sequential regorafenib, but also an effective therapeutic target whose inhibition enhanced regorafenib sensitivity after SR or LR.

Objective:To investigate the inhibitory effect of probiotics on cholesterol gallstone formation in mice fed a high-fat diet and its impact on leptin and intestinal flora.Methods:Forty 8-week-old female C57BL/6J SPF mice were obtained. After one week of adaptive feeding, the mice were randomly assigned to G-NS group ( n=10), G-Probiotics group ( n=10), L-NS group ( n=10), and L-Probiotics group ( n=10). The G-NS group and G-Probiotics group were fed with standard diet, while the L-NS group and L-Probiotics group received lithogenic diet. Additionally, the G-Probiotics group and L-Probiotics group received a probiotic solution (5×10 9 CFU/ml, 0.1 mL/10 g) by gavage, once daily for 8 weeks. The G-NS group and L-NS group received an equal volume of normal saline by gavage. After 8 weeks, the stone formation rate of mice was observed, the serum leptin, total cholesterol (TC) and low-density lipoprotein cholesterol (LDL) contents of mice were detected, feces were collected and DNA was extracted to analyze the diversity and abundance of intestinal flora in mice. Count data were presented as percentages, Fisher′s exact probability test was used for comparisons, and measurement data with normal distribution were presented as mean±standard divation ( ± s). One-way ANOVA was applied for inter-group comparisons, and the K-W rank sum test was utilized to analyze inter-group differences in the intestinal microbiota section. Results:No gallstone was found in the G-NS group and G-Probiotics group. The stone formation rates of the L-NS group and the L-Probiotics group were 100% and 60%, respectively, with statistical significance ( P<0.05). In the G-NS group, L-NS group, G-Probiotics group and L-Probiotics group the serum total cholesterol (TC) content were (2.03±0.34) mmol/L, (4.75±0.76) mmol/L, (1.64±0.49) mmol/L and (3.66±0.62) mmol/L, the serum low-density lipoprotein (LDL) cholesterol content was (0.57±0.10) mmol/L, (1.55±0.29) mmol/L, (0.73±0.37) mmol/L and (1.06±0.16) mmol/L, the serum leptin content was (6.77±0.76) μg/L, (19.24±3.97) μg/L, (3.21±1.32) μg/L and (11.67±1.05) μg/L. Comparison of serum TC and LDL: L-NS group was higher than G-NS group, L-NS group was higher than L-Probiotics group, L-Probiotics group was higher than G-Probiotics group, the difference was statistically significant ( P<0.01). Comparison of serum leptin showed that L-NS group was higher than G-NS group and L-Probiotics group, G-NS group and L-Probiotics group was higher than G-Probiotics group, the difference was statistically significant ( P<0.01). Compared with the control group, the intestinal flora diversity of mice in the simple high-fat diet group decreased, the relative abundance of harmful bacteria increased, and the relative abundance of beneficial bacteria decreased. After probiotics intervention, the intestinal flora diversity increased, the relative abundance of beneficial bacteria increased, and the relative abundance of harmful bacteria decreased. Conclusion:Probiotics can lower leptin levels and alleviate leptin resistance, decrease serum TC and LDL levels, and reduce cholesterol levels in bile, thus reducing cholesterol buildup in the gallbladder and preventing stone formation.

Objective To investigate the effect of microRNA-21 (miR-21) antisense oligonucleoti-de on collagen synthesis in the rat hepatic satellite cells ( HSCs) .Methods Rat hepatic stellate cells were isolated and cultured; the miR-21 antisense oligonucleotide was transfected into HSCs with lipofectamine 2000;after incubation 48 h, the HSCs were collected.The expression of miR-21 was detected with reverse transcription polymerase chain reaction ( RT-PCR) , andα-smooth muscle actin (α-SMA) protein in HSCs with Western blot.The cell proliferation was assayed with methyl thiazolyl tetrazolium ( MTT) method.Re-sults Compared to scrambled control group, the expression of miR-21 was reduced by 76%( P <0.01), the proliferation activity of HSCs was reduced by(26 ±3)%( P <0.01), the expressions of type I and III collagen proteins were reduced by(61 ±7)%and (48 ±6)%( P <0.01).Conclusions The miR-21 an-tisense oligonucleotide could reduce significantly the expression of miR-21, and inhibit HSC proliferation and extracellular matrix production.

Objective To study the diagnostic value of common inflammatory markers in patients with infectious diseases. Methods One hundred sepsis patients, 100 viral infection patients,100 pulmonary tuberculosis patients and 100 gonorrhea patients were analyzed retrospectively. The contents of procalcitonin (PCT), C-reactive protein (CRP), haptoglobin (HP), ceruloplasmin (CER), α1-acid glycoprotein (α1-AAG), α1-antitrypsin (α1-AAT), white blood cell count (WBC) and erythrocyte sedimetation rate (ESR) were measured. The receiver operating characteristic curve (ROC curve), sensitivity, specificity, positive predictive value, negative predictive value, Youden's index,positive and negative likelihood ratios and total coincidence rate were calculated respectively. Results The area under the ROC curve, sensitivity, specificity, Youden's index and positive likelihood ratios,positive predictive value and total coincidence rate of PCT in sepsis patients were 0. 895, 0.84, 0.92,0.76, 10.50, 0.91 and 0.88, respectively, which were superior to CRP, HP, CER, α1-AAG, α1-AAT, WBC and ESR. Conclusions PCT is a better inflammatory reactive parameter than other parameters currently applied in practice and may serve as a rapid and sensitive test in the early stage of severe bacterial infections.

Objective To investigate the effect of TGFβ1 siRNA on hepatic satellite cells (HSCs) activation, proliferation and extracellular matrix production. Methods The TGFβ1, siRNA plasmid was transfected into HSCs with Lipofectamine 2000. The supernatant and HSCs were collected after incubation for 72h. The expression of TGFβ1, and a-SMA protein in HSCs was detected by. Western blotting. The expression of type Ⅰ and Ⅲ collagen mRNA was detected by RT-PCR. The cell proliferation was assayed by MTT method. Contents of typeⅣ collagen and hyaluronic acid in supernatant were determined by radioimmuno-assay.Results Compared with scrambled control group, the TGFβ1, and a-SMA protein expression,the activity of HSCs proliferation,the expression of typeⅠ and Ⅲ collagen mRNA,and the contents of type Ⅳ collagen and hyaluronic acid in supernatant were reduced in TGFβ1, siRNA group by (79±5)%,(55±4)%, (25±4)% ,(63±6)% ,(57±4)% ,(53±8)% ,(46±8)% ( P<0.01),respectively.Conclusion TGFβ1, siRNA could significantly reduce the expression of TGFβ1,inhibited HSC activation,proliferation and extracellular matrix production.