Acute liver injury (ALI) serves as a critical precursor and major etiological factor in the progression and ultimate manifestation of various hepatic disorders. The prevention and treatment of ALI is still a serious global challenge. Given the limited therapeutic options for ALI, exploring novel targeted therapeutic agents becomes imperative. The potential therapeutic efficacy of inhibiting RIPK2 is highlighted, as it may provide significant benefits by attenuating the MAPK pathway and NF-κB signaling. Herein, we propose a CMD-OPT model, a two-stage molecular optimization tool for the rapid discovery of RIPK2 inhibitors with optimal properties. Compound RP20, which targets the ATP binding site, demonstrated excellent kinase specificity, ideal oral pharmacokinetics, and superior therapeutic effects in a model of APAP-induced ALI, positioning RP20 as a promising preclinical candidate. This marks the first application of RIPK2 inhibitors in ALI treatment, opening a novel therapeutic pathway for clinical applications. These results highlight the efficacy of the CMD-OPT model in producing lead compounds from known active molecules, showcasing its significant potential in drug discovery.

ObjectiveTo analyze the temporal trend of knockdown resistance (kdr) gene mutations highly correlated with pyrethroid resistance in field populations of Aedes aegypti in Jinghong City of Yunnan Province, and to provide a scientific basis for formulating rational insecticide use strategies. MethodsAdult mosquito samples of Aedes aegypti from 2016 to 2023 and larvae mosquito samples from July 2022 to June 2023 were collected in Jinghong City of Yunnan Province. Allele specific PCR (AS-PCR) was used to measure kdr mutations at amino acid positions 989, 1016 and 1534 of the voltage-gated sodium ion channel (VGSC) gene. Data such as mutation rate and mutation allele frequency were calculated, SPSS software was used to perform trend chi square tests on mutation rate and mutation allele frequency with year and month, as well as comparison of mutation allele frequencies and genotype distributions between the dry and rainy seasons, thereby delineating the temporal trend of kdr gene mutations. ResultsAmong the 173 samples collected from 2016 to 2023, the mutation rates of S989P and V1016G were 100.00% for each year, while the mutation rate of F1534C ranged from 62.50% to 100.00%. The mutation rate and mutation allele frequency of F1534C were increased over the years (χ²=22.079, P<0.001; χ²=42.971, P<0.001). Concurrently, the proportion of the PPGGCC genotype was increased annually (χ²=60.790, P<0.001). Among the 288 samples collected from July 2022 to June 2023, the monthly mutation rates for S989P, V1016G, and F1534C were consistently 100.00%. There was only one type of mutation present, namely S989P+V1016G+F1534C. In the combinations of the three genotypes, the SPGGCC genotype accounted for 1.39% (4/288), the PPGGFC accounted for 2.78% (8/288), and the PPGGCC had the highest proportion at 95.83% (276/288). After tesiting the samples collected in August 2023, the mutation rates of 989, 1016 and 1534 sites of VGSC in females, males, and larvae of the same generation were all 100.00%. ConclusionSince 2016, the gene mutations at S989P and V1016G loci in the VGSC gene of wild Aedes aegypti in Jinghong City have remained consistently at 100.00%, while the mutation rate and mutant allele frequency of F1534C have increased year by year during the testing period. By 2023, the mutation rates at three loci in the VGSC gene of Aedes aegypti in Jinghong City had all reached 100.00%, and neither changes in insect developmental stage nor gender differences during transmission exerted a detectable impact on the mutation rates. In the control of Aedes aegypti in Jinghong City, the use of pyrethroid insecticides should be stopped or reduced, and regular monitoring of kdr genes should be carried out to promptly detect new mutations.

ObjectiveTo investigate the mechanism of Jiedu Huoxue prescription in promoting the reendothelialization of injured vessels by regulating the nuclear factor （NF）-κB/NOD-like receptor protein 3 （NLRP3）/cysteine-aspartic acid protease （Caspase）-1-mediated pyroptosis. MethodA rat model of injured thoracic aorta was established by balloon injury， and 36 rats were assigned into shame surgery， model， low-， medium-， and high-dose Jiedu Huoxue prescription， and atorvastatin calcium tablet groups. The injured aortic segment was collected 28 days after surgery. Hematoxylin-eosin （HE） staining and Evans blue staining were conducted to reveal the changes of vascular structural morphology and the reendothelialization of blood vessels， respectively. The enzyme-linked immunosorbent assay （ELISA） was employed to determine the levels of tumor necrosis factor-α （TNF-α）， intercellular adhesion molecule-1 （ICAM-1）， interleukin （IL）-1β， and nitric oxide （NO） in the serum. Western blotting was employed to determine the expression of endothelial nitric oxide synthase （eNOS）， NF-κB p65， phospho-NF-κB p65 （p-NF-κB p65）， NLRP3， and Caspase-1 in the vascular tissue. ResultThe model group showed thickened endovascular membrane， proliferation and disarrangement of smooth muscle cells of the artery wall， obvious inflammatory cell infiltration， and narrowed luminal area. Jiedu Huoxue prescription and atorvastatin calcium tablets mitigated the pathological changes of the thoracic aorta in different degrees. After balloon injury， the endothelial coverage rate of the model group decreased significantly， while Jiedu Huoxue prescription and atorvastatin calcium tablets increased the reendothelialization rate （P<0.05）. Compared with the shame surgery group， the model group showed elevated levels of TNF-α， ICAM-1， and IL-1β （P<0.01） and lowered NO level （P<0.01） in the serum. In addition， the model group presented down-regulated protein level of eNOS （P<0.01） and up-regulated phosphorylation of pyroptosis-associated proteins NLPR3， Caspase-1， and NF-κB p65 in the vascular tissue （P<0.05， P<0.01）. Compared with the model group， Jiedu Huoxue prescription and atorvastatin calcium tablets lowered TNF-α， ICAM-1， and IL-1β levels （P<0.05， P<0.01） and elevated the NO level in the serum （P<0.05， P<0.01）. Moreover， the drugs up-regulated the expression of eNOS （P<0.01） and down-regulated the expression of NLRP3， Caspase-1， and NF-κB p65 （P<0.05， P<0.01） in the vascular tissue. ConclusionJiedu Huoxue prescription can promote the reendothelialization and inhibit the intimal hyperplasia of vessels after balloon injury by regulating the NF-κB/NLRP3/Caspase-1 pathway to inhibit pyroptosis and reduce endothelial inflammatory injury.

Objective : By testing the tensile strength of the poly(vinyl alcohol) (PVA)/bacterial cellulose(BC) composite membrane and its effect on the proliferation of mouse embryonic fibroblasts, its potential as new bone tissue engineering membrane were studied. Methods : PVA⁃BC films of different proportions and pure PVA films were prepared by self⁃evaporation method. The tensile strength of each group was tested. The group with the highest tensile strength was immersed in deionized water for 0. 5 h to measure its wet tensile strength. The microstructure of pure PVA film and the film with the highest tensile strength was observed by scanning electron microscopy (SEM) .X⁃ray diffraction and Fourier transform infrared ( FTIR) spectroscopy were used to analyze pure PVA, pure BC,and the film with the highest tensile strength respectively. Cell counting kit⁃8 (CCK⁃8) was applied to detect the survival rate in the blank control group, the pure PVA film group, and the composite film group with the highest tensile strength. Results : PVA⁃BC composite films were successfully prepared, X⁃ray diffraction and FTIR analysis revealed the co⁃presence of PVA and bacterial cellulose in the composite film. The initial tensile strength of the composite membrane increased with the BC ratio. When the concentration ratio of PVA to BC was 10 ∶ 7, the tensile strength reached (155. 5 ± 14. 7) MPa, and wet samples reached (13. 8 ± 1. 2) MPa. The CCK⁃8 test of NIH/3T3 showed that there was no significant difference among the PVA⁃BC composite film group, pure PVA group and blank control group after 1,4 and 7 days of cell culture ( P > 0. 05 ) . Conclusion PVA⁃BC film fabricated by blending method obtain certain mechanical properties and biocompatibility in both wet and dry states, which may be an appropriate candidate as a GBR membranes for clinical application.

Objective : To explore the feasibility of CS-BC composite membrane as guided bone regeneration(GBR) membrane by preparing chitosan(CS)-bacterial cellulose(BC) composite membrane, and testing the mechanical strength of the composite membrane both in dry and wet states as well as evaluating its cytocompatibility. Methods: The CS solution and BC solution were mixed in different weight ratios and were dispersed evenly by ultrasonic. Pure CS membrane and composite membranes with different weight ratios of CS and BC(10 ∶1, 10 ∶3, 10 ∶5, 10 ∶7 and 10 ∶9) were prepared by self-evaporation process. Thereafter, the fabricated membranes were immersed into sodium hydroxide ethanol solution to remove the acid. The tensile strength of the as-fabricated membranes under dry condition and under hydration were measuredviamechanical universal testing machine(n=6). The microstructures of the composite membrane with the highest tensile strength were observed by scanning electron microscopy(SEM) and transmission electron microscopy(TEM). To characterize the chemical composition of the composite membrane, Fourier transform infrared(FTIR) spectrometer and X-ray diffraction(XRD) were used. CCK-8 assay was carried out to evaluate the survival rate of cells in the control group(without any membrane), the pure CS membrane group and the composite membrane group with maximum tensile strength(n=5) after cocultured with rat bone marrow stem cells(RBMSCs) for 1, 4 and 7 d. Results : The cross-section of the composite membrane displayed ordered layer structure after introducing BC into CS matrix. The results of FTIR and XRD indicated the existence of BC in the composite membrane. The tensile strength of the CS-BC composite membrane increased first and then decreased with the increase of BC ratio. The tensile strength of the composite membrane reached the highest in dry and wet states when the weight ratio of CS and BC is 10 ∶7, which were almost(204.7±63.0) MPa and(44.4±6.4) MPa respectively. After cocultured with RBMSCs for 1, 4 and 7 d, there was no significant difference in the number of cells among the pure CS group, the CS-BC composite membrane group and the blank control group. Conclusion When the weight ratio of CS and BC is 10 ∶7, the tensile strength of the composite membrane are the best both in dry and wet state and the cell compatibility is excellent.

Objective: To investigate the effects of nutritional interventions based on nutritional risk assessment strategies on nutritional status and liver function in patients with cirrhosis. Methods: Totally126 patients with cirrhosis admitted from July 2016 to October 2017 were enrolled. The prospective study was used and the patients were randomly divided into the treatment group and the control group with 63 cases in each group. The control group received routine nursing intervention, while the treatment group was combined nutritional intervention based on the nutritional risk assessment strategy. After 3 months of follow-up, serum nutrition indicators, liver function, and complications were compared between the two groups. Results: The serum albumin, prealbumin and transferrin levels in the treatment group were (42.36±5.24) g/L, (0.37±0.08) g/L, and (3.01±0.42) g/L, which were significantly higher than those in the control group, which were (34.24±5.36) g/L, (0.30±0.07) g/L, (2.65±0.52) g/L (t=8.598, 5.227, 4.275, P<0.05 or 0.01); serum alanine aminotransferase and total bilirubin content were (95.32±12.45) μmol/L, (272.36±30.12) μmol/L, which were significantly lower than those of the control group [(114.21±13.20) μmol/L, (320.42±41.52) μmol/L]. The prothrombin time [(14.36±1.32) s] was significantly shorter than that of the control group [(16.12±1.20) s] (t=8.263, 7.437, 7.831, P<0.05); the complication rate of the treatment group was 7.94% (5/63), which was significantly lower than that of the control group (22.22%) (14/63)(χ²=5.020, P<0.05). Conclusions Nutrition intervention based on nutritional risk assessment strategy can improve nutritional status, relieve liver damage and reduce the incidence of complications such as peritonitis in liver cirrhosis patients.

Gap junctions mediate direct communication between cells; however, toxicological cascade triggered by nonessential metals can abrogate cellular signaling mediated by gap junctions. Although cadmium (Cd) is known to induce apoptosis in organs and tissues, the mechanisms that underlie gap junction activity in Cd-induced apoptosis in BRL 3A rat liver cells has yet to be established. In this study, we showed that Cd treatment decreased the cell index (a measure of cellular electrical impedance) in BRL 3A cells. Mechanistically, we found that Cd exposure decreased expression of connexin 43 (Cx43), increased expression of p-Cx43 and elevated intracellular free Ca2+ concentration, corresponding to a decrease in gap junctional intercellular communication. Gap junction blockage pretreatment with 18β-glycyrrhizic acid (GA) promoted Cd-induced apoptosis, involving changes in expression of Bax, Bcl-2, caspase-3 and the mitochondrial transmembrane electrical potential (Δψm). Additionally, GA was found to enhance ERK and p38 activation during Cd-induced activation of mitogen-activated protein kinases, but had no significant effect on JNK activation. Our results indicated the apoptosis-related proteins and the ERK and p38 signaling pathways may participate in gap junction blockage promoting Cd-induced apoptosis in BRL 3A cells.
Animals ; Apoptosis/*drug effects ; Cadmium/*toxicity ; Calcium/metabolism ; Cell Communication/drug effects ; Connexin 43/genetics ; Enzyme Activation/drug effects ; Gap Junctions/*drug effects ; Gene Expression Regulation/drug effects ; Hepatocytes/cytology/*drug effects ; Rats ; Signal Transduction/drug effects

Objective To evaluate the effects of stepwise and fine-grained dietary management on the diet control of liver cirrhosis patients complicated with gastrointestinal hemorrhage. Methods Eighty liver cirrhosis patients complicated with gastrointestinal hemorrhage were assigned to experimental group (41 cases) and control group(39 cases) by random digits table method. The patients in experimental group were treated with stepwise and fine-grained dietary management and the patients in control group were given conventional dietary treatments. The compliance diet, mastery of nutrition knowledge, malnutrition risk at admission, intake food, discharge and 2 months after discharge between two groups were monitored. The incidence of rebleeding was tracked after 1 year of discharge. Results The incidence of malnutrition risk in Child-Pugh class C was higher than that in Child-Pugh class B( Z=-4.3, P<0.05 ) . The incidence of malnutrition risk in patients with high education level was lower than that in patients with lower education( r=-0.453, P<0.05 ). The experimental group significantly outperformed control group in mastery of nutrition knowledge at discharge:80(70, 84) points vs. 52(42, 64) points, in compliance diet and malnutrition risk after 2 months after discharge:86(76, 91)%vs. 53(46,57)%, 1(1, 2) points vs. 2(1,3) points, the rebleeding rate in experimental group was significantly less than that in control group within 1 year after discharge 14.6%(6/41) vs. 35.9%(14/39), and the differences were statistically significant between two groups ( χ2 = 4.226- 51.232, all P < 0.05 ). Conclusions Stepwise and fine-grained dietary management can improve the mastery of nutrition knowledge, compliance diet and nutritional status, and reduce the occurrence of rebleeding, worth popularizing further in clinical care of liver cirrhosis patients complicated with gastrointestinal hemorrhage.

Objective To evaluate the effect of neoadjuvant chemotherapy (NAC) by investigating the expression of cyclin D1 in human breast cancer before and after NAC.Methods Eighty-four cases of breast cancer were diagnosed by core biopsies.The expression level of cyclin D1 in cancer tissues was measured by immunohistochemical envision two-step method before and after NAC (pirarubicin and docetaxel regimen for 3-4 cycles).Results Complete remission (CR) occurred in 4 cases of 84 patients (4.76 ％) with 2 pathological complete response cases,partial response (PR) in 54 cases (64.29 ％),stability (SD) in 26 cases (30.95 ％) and no disease progression (PD) patients.The positive rate of cyclin D1 in cancer tissues [65.48 ％ (55/84)] was significantly decreased after NAC [39.29 ％ (33/84)] (x2 =11.55,P =0.001).In clinical level,the ease rate was significantly improved in patients whose cyclin D1 expression switched from positive [86.36 ％ (19/22)] to negative [45.45 ％ (15/33)] after NAC treatment (x2 =9.359,P =0.002).Conclusion NAC significantly decreases the expression of cyclin D1 in breast cancer tissues.Meanwhile,the ease rate is improved when cyclin D1 expression switched from positive to negative after NAC.Therefore,cyclin D1 expression can be used as an evaluation index for the efficiency of NAC.

OBJECTIVETo study the effects of different compatibility proportion of Jiaotai pills on treating type 2 diabetes mellitus (T2DM) in rats.

METHODThe model of type 2 diabetes mellitus in rats was established by injecting streptozotocin from tail vein and feeding with high fat and high caloric diet. Diabetic rats were randomly divided into model group, Jiaotai pill 1 group (Coptidis Rhizoma-cinnamon 2: 1), Jiaotai pill 2 group (Coptidis Rhizoma-cinnamon 4: 1), Jiaotai pill 3 group (Coptidis Rhizoma-cinnamon 10: 1) and metformin group. Rats in different treatment groups were given by corresponding therapy from gastric tube. Meanwhile normal control group was another set. Body weight, oral glucose tolerance test (OGTT), blood lipid level including total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C), plasma levels of free fatty acid (FFA) and adiponectin, plasma liver enzymes activity(ALT, AST, AKP, gamma-GT) and pathological results of liver tissue were determined after eight weeks.

RESULTBody weight, fasting plasma glucose (FPG), postpradial plasma glucose at one hour (PG-1 h), postpradial blood glucose at two hour (PG-2 h), plasma levels of TC, TG, LDL-C, FFA and liver enzymes activity were all increased in rats of model group compared with those in normal control group. Plasma levels of HDL-C and adiponectin were decreased in model group (P < 0.01). Fatty degeneration of hepatocytes was apparent in liver tissues in rats of model group. Compared with model group results of OGTT, blood lipid levels and liver enzymes activity were improved while levels of HDL-C and adiponectin were increased in rats of different treatment groups (P < 0.05 or P < 0.01). Meanwhile fatty degeneration of hepatocytes was improved in liver tissues in rats of different treatment groups. Compared with metformin group, plasma level of HDL-C was elevated while AKP and gamma-GT were decreased significantly in rats of Jiaotai pill 1 group (P < 0.05), gamma-GT level was decreased significantly in rats of Jiaotai pill 2 group (P < 0.05), AST, AKP and gamma-GT levels were decreased significantly in rats of Jiaotai pill 3 group (P < 0.05). Compared with Jiaotai Pill 1 group, plasma levels of HDL-C was decreased while AKP levels was elevated significantly in rats of Jiaotai pill 2 group, but HDL-C was decreased in rats of Jiaotai pill 3 group (P < 0.05).

CONCLUSIONIt is suggested that different compatibility proportion of Jiaotai pills are effective on treating type 2 diabetes mellitus in rats. The effect of Jiaotai pill 1 group is better than that of other therapy groups.

Animals ; Blood Glucose ; analysis ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Acids, Nonesterified ; blood ; Lipids ; blood ; Male ; Medicine, Chinese Traditional ; Rats ; Streptozocin