Objective To address the quality problems caused by high porosity in the preparation of dental cobalt-chrome alloy prosthetics based on selective laser melting(SLM)technology,we investigated the influence mechanism of different forming process parameters on the microstructure and properties of the materials.Moreover,the range of form-ing process parameters that can effectively reduce defects was precisely defined.Methods The effects of laser power,scanning speed,and scanning distance on the pore properties,surface roughness,and hardness of dental cobalt-chrome al-loy were investigated by adjusting the printing parame-ters in the process of SLM.Through metallographic anal-ysis,image analysis,and molten pool simulation,the pore formation mechanism was revealed,and the relation-ship between the porosity and energy density of SLM dental cobalt-chrome alloy was elucidated.Results When the linear energy density was higher than 0.18 J/mm,the po-rosity defect easily appeared at the bottom of the molten pool.When the laser energy density was lower than 0.13 J/mm,defects occurred in the gap of the molten pool due to insufficient melting of powder.In particular,when the linear energy density exceeded the threshold of 0.30 J/mm or was below 0.12 J/mm,the porosity increased significantly to more than 1%.In addition,we observed a negative correlation between free surface roughness and energy density and an inverse re-lationship between macroscopic hardness and porosity.Conclusion On the basis of the conditions of raw materials and molding equipment used in this study,the key process parameters of SLM of molding parts with porosity lower than 1%were successfully determined.Specifically,these key parameters included the line energy density,which ranged from 0.13 J/mm to 0.30 J/mm,and the scan spacing should be strictly controlled below 90 μm.

Objective:To explore surgical strategies for acute type A aortic dissection involving severe stenosis or occlusion of the carotid arteries.Methods:From January 2019 to March 2023, a total of 29 patients with acute type A aortic dissection involving severe stenosis or occlusion of the carotid arteries were included in the study. All patients underwent emergency surgery, with simultaneous intraoperative neck incision and replacement of the unilateral or bilateral carotid arteries. Among them, there were 19 males with a mean age of(49.57±2.14)years old. Preoperative brain CT indicated abnormalities in 15 cases, transient neurological dysfunction occurred in 5 cases, and syncope in 1 case.Results:Procedures included ascending aorta replacement in 10 cases, Bentall procedure in 18 cases, and Wheat procedure in 1 case. Arch operations involved partial arch replacement in 3 cases and Sun’s procedure in 26 cases. Simple left carotid artery replacement was performed in 6 cases, simple right carotid artery replacement in 19 cases, and bilateral carotid artery replacement in 4 cases. Cerebral protection measures during circulatory arrest included unilateral cerebral perfusion in 24 cases and bilateral cerebral perfusion in 5 cases. The mean operation time was(7. 6±0. 3) h, with a mean cardiopulmonary bypass time of(196. 3±8. 7) min, aortic cross-clamp time of(113.2±6.4) min, ischemic time 12(5-16.5) min, and lowest temperature of(26.3±0.4)°C. One patient experienced in-hospital mortality. Postoperatively, new neurological dysfunction occurred in 2 cases, including 1 case with coma and permanent neurological deficit.Conclusion:In patients with acute type A aortic dissection involving severe stenosis or occlusion of the carotid arteries, simultaneous carotid artery replacement via neck incision during aortic surgery is a safe and reliable surgical approach.

miR-135 is a highly conserved miRNA in mammals and includes miR-135a and miR-135b.Recent studies have shown that miR-135b is a key regulatory factor in cardio-cerebrovascular diseases.It is involved in regulating the pathological process of myocardial infarction,myocardial ischemia/reperfusion injury,cardiac hypertrophy,atrial fibrillation,diabetic cardiomyopathy,atherosclerosis,pulmonary hyperten-sion,cerebral ischemia/reperfusion injury,Parkinson's disease,and Alzheimer's disease.Obviously,miR-135b is an emerging player in cardio-cerebrovascular diseases and is expected to be an important target for the treatment of cardio-cerebrovascular diseases.However,the crucial role of miR-135b in cardio-cerebrovascular diseases and its underlying mechanism of action has not been reviewed.Therefore,in this review,we aimed to comprehensively summarize the role of miR-135b and the signaling pathway mediated by miR-135b in cardio-cerebrovascular diseases.Drugs targeting miR-135b for the treatment of diseases and related patents,highlighting the importance of this target and its utility as a therapeutic target for cardio-cerebrovascular diseases,have been discussed.

Objective:To predict and analyze the T/B combined epitope of EM10 protein in Echinococcus multilocularis, and identify the expressed products of the biosynthetic EM10 multi epitopes. Methods:The gene-related information of EM10 protein was obtained through NCBI GenBank public database. Bioinformatics technique was used to predict and analyze the T/B binding epitopes of EM10 protein. The prokaryotic expession recombinant plasmid pET30a-EM10 (epitope) was synthesized, and transformed into host bacteria Ecoli. BL21 (DE3). The expression of EM10 recombinant multi-epitope protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting after induced expression by isopropyl thiogalactopyranoside (IPTG). Results:The total length of EM10 gene was 1 759 bp (GenBank registration number: U05573), and its protein amino acid sequence (GenBank registration number: AAA50580.1) was 559 amino acids. By using Phyre software for homology modeling, the tertiary structure of EM10 protein was obtained, and the T/B combined epitope of EM10 protein was successfully predicted, the dominant epitope was located at 46 - 61, 133 - 183, 239 - 255 and 442 - 475 amino acid sites. The (GGGGS)n linker sequence was used to connect the epitopes to form an EM10 recombinant multi-epitope protein with a total of 206 amino acid. The size of the DNA fragment was 618 bp and the relative molecular weight of the protein was 22.66 × 10 3. The prokaryotic expession recombinant plasmid was validated by enzyme digestion, the results showed that the plasmid size was between 5 000 and 6 000 bp, which was consistent with the length of the constructed plasmid (5 854 bp). SDS-PAGE showed that the target protein was expressed in the supernatant induced by IPTG at 37 ℃ and the effect was the best. The relative molecular weight of the protein was 22.66 × 10 3 by Western blotting, which was consistent with the constructed plasmid. Conclusions:The combined epitope of EM10 T/B is successfully designed and predicted using bioinformatics technology. A prokaryotic expression recombinant plasmid is constructed, the expression of EM10 recombinant multi-epitope protein is verified through experiments, providing an experimental basis for the construction of an EM10 dominant epitope diagnostic kit.

Objective : To investigate the drug resistance and pathogenicity of six clinical isolates of Klebsiella pneu- moniae (Kp) ，and to provide a basis for prevention and treatment of Kp infection． Methods : The six strains from different hospitals were isolated ，cultured ，and identified by species-specific gene khe． Their whole genome se- quences (WGS) were obtained using next-generation sequencing technology (NGS) ．Based on the WGS，the cap- sular serotypes，sequence types (ST) and drug-resistance genes of six strains were identified．The capsular sero- type genes and virulence genes were validated or identified using PCR. Broth microdilution tests were conducted to validate their drug susceptibility，and mice were challenged with Kp aerosols by MicroSprayer aerosolizer to evaluate their pathogenicity. Results : The six strains were all serotype K2 but belonged to four ST types ( ST14 ，ST65， ST700，and ST86) ，and collectively carried six virulence genes and 23 drug-resistance genes．All the six strains were resistant to ampicillin，but only one strain was multidrug-resistant．Four strains exhibited high mucoid charac- teristics．Five strains could cause mortality in mice，which were preliminary identified as high virulence strains. Conclusion For the six Kp clinical isolates from different sources，only one strain named NY 13294 is both multi- drug-resistant and highly virulent，and other four highly virulent strains are resistant to one or two types of antibiot- ics.

Diabetes mellitus (DM) is a major metabolic disease endangering global health, with diabetic nephropathy (DN) as a primary complication lacking curative therapy. Sporoderm-broken spores of Ganoderma lucidum (GLP), an herbal medicine, has been used for the treatment of metabolic disorders. In this study, DN was induced in Sprague-Dawley rats using streptozotocin (STZ) and a high-fat diet (HFD), and the protective mechanisms of GLP were investigated through transcriptomic, metabolomic, and network pharmacology (NP) analyses. Our results demonstrated that GLP intervention ameliorated renal damage and inflammation levels in DN rats. Integrative metabolomic and transcriptomic analysis revealed that GLP treatment modulated glucose and cellular energy metabolisms by regulating relevant genes. GLP significantly suppressed the inflammations by impacting glucose and energy metabolism-related gene expression (Igfbp1 and Angptl4) and enhanced metabolic biomarkers of 4-Aminocatechol. In addition, NP analysis further indicated that GLP may efficiently alleviate DN via immune-related pathways. In conclusion, this study provides supportive evidence of the anti-inflammatory effects of GLP supplements, highlighting their potential for promising clinical applications in treating DN.

Urate oxidase (Uox) plays a pivotal role in uric acid (UA) degradation, and it has been applied in controlling serum UA level in clinical treatment of hyperuricemia (HUA). However, because Uox is a heterogenous protein to the human body, the immune rejections typically occur after intravenous administration, which greatly hampers the application of Uox-based agents. In this study, we used Lactococcus lactis NZ9000, a food-grade bacterium, as a host to express exogenous Uox genes, to generate the Uox-expressing engineered strains to treat HUA. Aspergillus flavus-derived Uox (aUox) and the "resurrected" human-derived Uox (hUox) were cloned into vector and expressed in NZ9000, to generate engineered strains, respectively. The engineered NZ9000 strains were confirmed to express Uox and showed UA-lowering activity in a time-dependent manner in vitro. Next, in an HUA mice model established by oral gavage of yeast paste, the UA levels were increased by 85.4% and 106.2% at day 7 and day 14. By contrast, in mice fed with NZ9000-aUox, the UA levels were increased by 39.5% and 48.3% while in mice fed with NZ9000-hUox were increased by 57.0% and 82.9%, suggesting a UA-lowering activity of both engineered strains. Furthermore, compared with allopurinol, the first-line agent for HUA treatment, mice fed with NZ9000-aUox exhibited comparable liver safety but better kidney safety than allopurinol, indicating that the use of engineered NZ9000 strains not only alleviated kidney injury caused by HUA, but could also avoided the risk of kidney injury elicited by using allopurinol. Collectively, our study offers an effective and safe therapeutic approach for HUA long-term treatment and controlling.
Animals ; Lactococcus lactis/metabolism* ; Urate Oxidase/genetics* ; Mice ; Uric Acid/blood* ; Hyperuricemia ; Humans ; Administration, Oral ; Aspergillus flavus/genetics* ; Male

OBJECTIVE to explore the material basis and potential molecular mechanism of Ziziphi Spinosae Semen- Acori Tatarinowii Rhizoma in the treatment of insomnia using network pharmacology and molecular docking, and establish insomnia mouse model by p-chlorophenylalanine(PCPA) for verification in vivo.METHODS Firstly, the chemical constituents of Ziziphi Spinosae Semen and Acori Tatarinowii Rhizoma were collected through TCMSP database and the potential active constituents were screened. The genes related of insomnia were obtained from GeneCards, OMIM and TTD databases, and the intersection targets of Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma and insomnia diseases were obtained. Then the network map of Chinese medicine-compound-target-disease was constructed in Cytoscape 3.6.1 software. The protein interaction network diagram was established in the STRING database. Functional enrichment analysis of target GO and KEGG pathways were performed using the DAVID database. Then the molecular docking technology was used for preliminary verification. The 60 male ICR mice were randomly divided into normal group, model group, high, medium and low dose groups(8.0, 4.0, 2.0 g·kg-1) and diazepam group(3 mg·kg-1). In addition to the normal group, PCPA(30 mg·mL-1) was intraperitoneally injected on the first and second days to establish the insomnia model. Then the drug was administered continuously for 7 d, and the normal group and the model group were given the same volume of normal saline. The sleep latency and duration of mice induced by the upper threshold dose of pentobarbital sodium(55 mg·kg-1), vertical and horizontal scores in the behavioral open-field experiment, open arm entry(OE%) and open arm time(OT%) of elevated cross maze were determined. HE staining was used to observe the hypothalamic histopathological situation. Serum levels of TNF-α and CASP3 were detected by ELISA. Finally, Western blotting was used to detect the expression of AKT1, p-AKT1 protein in the hypothalamus of the mice in each group. RESULTS The potential active components of Ziziphi Spinosae Semen and Acori Tatarinowii Rhizoma were 9 and 5, and the common targets with insomnia were 34. A total of 160 GO items were obtained through GO enrichment analysis. KEGG pathway analysis found that the signaling pathway was mainly related to inflammatory signaling pathway, among which AKT1, CASP3 and TNF were the key targets. The results of molecular docking showed that the selected compounds had high binding activity with the key targets. Animal experiment results showed that the Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma for insomnia could significantly shorten the model mice sleep latency, prolong sleep duration, reduce the vertical and horizontal score, improve the OE% and OT%, restore the hypothalamus pathological tissue damage, significantly reduce the content of TNF-α and CASP3, raise the level of AKT1 protein expression in the tissue of the hypothalamus. CONCLUSION Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma can regulate TNF signaling pathway by acting on TNF-α, CASP3, AKT1, p-AKT1 and other targets to treat insomnia.

Malnutrition in liver cirrhosis is associated with ascites, hepatic encephalopathy, infection, and other complications even death. To date, the hazard and disease burden of malnutrition in cirrhosis patients have been severely underestimated. This review summarized the most recent advancement in the field and discussed the techniques and methodologies in detection and evaluation of malnutrition in cirrhosis patients, nutritional support therapy, and future research directions and clinical care of the patients.

OBJECTIVE: To assess the influence of rs2910164 G/C single nucleotide polymorphism (SNP) of the miR-146a gene on its expression and susceptibility to gastric cancer. METHODS: Fifty three gastric cancer patients and six gastric cancer cell lines were selected for determining the miR-146a expression by Taqman quantitative PCR. A model was constructed to assess the influence of miR-146a overexpression on the growth of AGS gastric cancer cells. A case-control study involving 417 gastric cancer patients and 420 cancer-free individuals was then conducted, and the allelic and genotypic frequencies of the rs2910164 G/C SNP were compared. The genotypes of all subjects were determined by using a Taqman allelic discrimination assay. A Taqman assay was also used to quantify mature and pri-miR-146a transcripts among 65 gastric cancer patients with known genotypes. RESULTS: The expression of miR-146a was down-regulated among the 53 gastric cancer patients and six gastric cancer cell lines. Over-expression of miR-146a has suppressed the growth of gastric cancer by inhibiting the G1/S-phase transition of AGS cells. The case-control study showed that subjects with GC/CC genotypes had significantly lower risk for gastric cancer compared with those with GG genotype. In addition, miR-146a G/C SNP has significantly increased the level of mature miR-146a in those with GC/CC genotype compared with GG genotype. CONCLUSION Down-regulation of miR-146a may play an important role in the pathogenesis of gastric cancer. The rs2910164 polymorphism of the miR-146a gene may reduce the risk of gastric cancer by influencing the processing of mature miR-146a.
Case-Control Studies ; Genotype ; Humans ; MicroRNAs/genetics* ; Polymorphism, Single Nucleotide ; Stomach Neoplasms/genetics*