Objective To evaluate the protective effect and underlying mechanism of ulinastatin on hepatic ischemia-reperfusion injury.Methods Twenty-four male SD rats were divided into three groups:sham operation group(Sham group),hepatic ischemia-reperfusion injury group(HIRI group)and hepatic ischemia-reperfusion injury+ulinastatin pretreatment group(HIRI+UTI group),with 8 rats in each group.The HIRI rat models were established by occluding hepatic portal vein and hepatic artery for 1 h.In the HIRI+UTI group,the rats were intraperitoneally injected with ulinastatin at 30 min before model establishment,and an equivalent amount of normal saline was given in the Sham and HIRI groups.The rats were sacrificed at 6 h after model establishment.Serum samples were collected to detect alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.Ultrastructural changes of mitochondria in liver tissues were observed by transmission electron microscopy.The expression of glutathione peroxidase 4(GPX4)was determined by immunofluorescent staining.The contents of malondialdehyde(MDA),glutathione(GSH),Fe,reactive oxygen species(ROS)and GPX4 were detected.The expression levels of GPX4 and acyl-CoA synthetase long-chain family 4(ACSL4)messenger RNA(mRNA)and proteins in liver tissue were measured.Results Compared with the Sham group,serum ALT and AST levels were up-regulated,pathological changes such as congestion,hepatocyte necrosis and abnormal hepatic lobule structure were observed,pathological score was increased,the mitochondria shrank,the membrane density was increased,the mitochondrial crest was damaged or even absent,the contents of ROS,MDA and Fe were elevated,the GSH content was decreased,the fluorescent intensity of GPX4 was weakened,the relative expression levels of ACSL4 mRNA and protein were up-regulated,and the relative expression levels of GPX4 mRNA and protein were down-regulated in the HIRI group(all P＜0.05).Compared with the HIRI group,serum ALT and AST levels were down-regulated,liver tissue injury was alleviated,pathological score was decreased,mitochondrial shrinkage and crest breakage were mitigated,the contents of ROS,MDA and Fe were down-regulated,the GSH content was up-regulated,the fluorescent intensity of GPX4 was enhanced,the relative expression levels of ACSL4 mRNA and protein were down-regulated,and the relative expression levels of GPX4 mRNA and protein were up-regulated in the HIRI+UTI group(all P＜0.05).Conclusions Ulinastatin may alleviate hepatic ischemia-reperfusion injury in rats probably through inhibiting ferroptosis.

Fecal microbiota transplantation(FMT)is a therapeutic approach that targets intestinal microorganisms by transplanting fecal microorganisms from healthy individuals into the gastrointestinal tract of diseased individuals,thus restoring the recipient's disordered gastrointestinal microbiota by restructuring the intestinal flora.However,the mechanism of action and adverse effects of FMT in different diseases have not yet been clarified,thus limiting its wide clinical application.Its use still relies on in-depth preclinical studies;however,highly inconsistent or incomplete experimental details provided in current reports,coupled with a lack of authoritative standards and recommendations,seriously affect the interpretation of the study findings and replication of the experimental procedures,as well as hindering the clinical translation of the result.We therefore review and discuss the key steps of recipient selection and graft sample collection,storage,graft material preparation,and grafting route,with the aim of improving the utilization of experimental animals,consumables,and labor,and providing method ological recommendations and references to achieve replicability and standardization of preclinical FMT studies.

OBJECTIVE To study the effect of different proportion compatibility of Astragali Radix and Chuanxiong Rhizoma on the extraction kinetics of flavonoids. METHODS The content determination method of flavonoids(with rutin as the control substance) was established, and the concentrations of flavonoids in the extracts of Astragali Radix membranicum and Chuanxiong Rhizoma in different proportions(1∶1, 1∶2, 1∶3, 1∶4, 1∶5, 2∶1, 3∶1, 4∶1, 5∶1) were determined dynamically within 2 h, respectively. The extraction kinetics model was established according to Feck's first diffusion law. The extraction rate constant was calculated and the difference of dissolution kinetics was compared. RESULTS The compatibility of different proportions of Astragali Radix and Chuanxiong Rhizoma had significant effects on the extraction rate and concentration of flavonoids. The extraction kinetics models of flavonoids were consistent with the characteristics of the first-order kinetic equation. The extraction rate was the fastest when Astragali Radix ∶ Chuanxiong Rhizoma was 3∶1, the extraction rate was the slowest when Chuanxiong Rhizoma∶ Astragali Radix was 2∶1. The equilibrium concentration of flavonoids was the highest when Chuanxiong Rhizoma∶ Astragali Radix was 3∶1, when the ratio of Astragali Radix∶Chuanxiong Rhizoma was 5∶1, it was the lowest. CONCLUSION The compatibility of different proportions of Astragali Radix and Chuanxiong Rhizoma has a significant effect on the extraction rate and concentration of flavonoids.

Objective:To investigate whether circular RNA 0026134 (circ_0026134) affects the radiosensitivity of hepatoma cells by regulating the miR-1270/growth factor receptor-bound protein 2 (GRB2) pathway.Methods:Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of circ_0026134, miR-1270, and GRB2 in liver cancer tissues and cells. Bioinformatics analysis, a dual-luciferase gene reporter assay, RT-qPCR, and western blot were used to analyze the targeting relationships between circ_0026134 and miR-1270 and miR-1270 and GRB2. The effects of circ_0026134, miR-1270, and GRB2 expression combined with 6 Gy on the proliferation, invasion, migration, and apoptosis of Huh7 and SK-HEP-1 cells were detected by a cell counting kit, a transwell assay, a scratch assay, and flow cytometry. The tumorigenesis experiment was used to detect the effect of silencing circ_0026134 in nude mice. Measurement data are expressed as the mean ± standard deviation. The independent sample t-test was used for comparison between two groups, and the one-way analysis of variance and SNK- q test were used for comparison between multiple groups. P < 0.05 was considered statistically significant. Results:The expression levels of circ_0026134 and GRB2, Huh7, and SK-HEP-1 cells in liver cancer tissues were significantly increased, while the expression levels of miR-1270 were significantly decreased ( P < 0.05). The expression of circ_0026134 in Huh7 and SK-HEP-1 decreased significantly after radiotherapy ( P < 0.05). circ_0026134 binds directly to miR-1270 and negatively regulates miR-1270 expression ( P < 0.05). miR-1270 binds directly to GRB2 and negatively regulates GRB2 expression ( P < 0.05). 6 Gy radiation significantly inhibited the proliferation, migration, and invasion of Huh7 and SK-HEP-1 cells and induced apoptosis ( P < 0.05). Silencing circ_0026134 or overexpression of miR-1270 significantly enhanced the anti-proliferation, anti-migration, invasion, and pro-apoptosis effects of 6 Gy treatment on hepatoma cells ( P < 0.05). Inhibition of miR-1270 significantly weakened the effects of silencing circ_0026134 combined with 6 Gy radiation on proliferation, migration, invasion, and apoptosis of hepatoma cells ( P < 0.05). Overexpression of GRB2 significantly weakened the effects of miR-1270 overexpression combined with 6 Gy radiation on proliferation, migration, invasion, and apoptosis of hepatoma cells ( P < 0.05). circ_0026134 knockdown significantly delayed tumor growth in vivo ( P < 0.05). Conclusion:Silencing circ_0026134 strengthens radiation treatment’s anti-proliferation, anti-migration, invasion, and pro-apoptotic effects in hepatoma cells by negatively regulating the miR-1270/GRB2 pathway, thereby enhancing radiosensitivity.

Objective:To explore the effects of the immune responses mediated by topological structures of three-dimensional bioprinted scaffolds on hair follicle cycle in mice.Methods:The study was an experimental research. The alginate-gelatin composite hydrogels were printed into scaffolds using a three-dimensional bioprinter and named T45 scaffolds, T60 scaffolds, and T90 scaffolds according to the 3 topological structures of the scaffolds (the rotation angles of the printhead during printing were 45°, 60°, and 90°, respectively), and the morphology of the three scaffolds was observed after cross-linking by naked eyes. Nine 8-week-old female C57BL/6J mice were divided into T45 group, T60 group, and T90 group, according to the random number table, with three mice in each group, and the T45, T60, and T90 scaffolds were subcutaneously implanted on the back of mice, respectively. On post implantation day (PID) 7, the hair growth in the dorsal depilated area of mice was observed, the thickness of the fiber capsule around the scaffolds was observed by hematoxylin-eosin staining, and the expression levels of CD68, bone morphogenetic protein-2 (BMP-2), and tumor necrosis factor (TNF) protein in the tissue surrounding the scaffolds were observed by immunofluorescence staining. The samples of the above experiments were all 3.Results:The topological structures of the three scaffolds were all clear with high fidelity after cross-linking. On PID 7, the hair growth was obvious in the dorsal depilated area of mice in T45 group and T90 group, while hair growth was slow in the scaffold implantation area of mice in T60 group, which was significantly different from that of the unimplanted area. On PID 7, compared with (18±4) μm in T90 group, the thickness of both the fiber capsule around the scaffolds ((39±4) and (55±8) μm) of mice in T45 group and T60 group was significantly increased ( P<0.05); the thickness of the fiber capsule around the scaffolds of mice in T60 group was also significantly increased compared with that in T45 group ( P<0.05). On PID 7, the expression level of CD68 protein in the tissue surrounding the scaffolds of mice in T60 group was significantly higher than the levels in T45 group and T90 group (with both P values <0.05). The expression level of BMP-2 protein in the tissue surrounding the scaffolds of mice in T60 group was significantly higher than the levels in T45 group and T90 group (with both P values <0.05), and the expression level of BMP-2 protein in the tissue surrounding the scaffolds of mice in T45 group was significantly higher than that in T90 group ( P<0.05). The expression level of TNF protein in the tissue surrounding the scaffolds of mice in T60 group was significantly lower than the levels in T45 group and T90 group (with both P values <0.05). Conclusions:Three-dimensional bioprinted scaffolds with different topological structures mediate different degrees of immune responses after being implanted in mice. A moderate immune response promotes hair growth in depilated area of mice, while an excessive immune response results inhibits the hair follicle entering into the anagen phase.

Objective:To analyze the predictive value of soluble cytotoxic T lymphocyte-associated antigen 4 (sCTLA-4) and RAD51 paralogous gene C (RAD51C) protein in the recurrence of cervical cancer patients after interventional therapy.Methods:A total of 107 patients with cervical cancer who underwent interventional surgery in our hospital from May. 2015 to Apr. 2019 were selected as the research group. postoperative recurrence were recorded. Another 107 patients with benign cervical disease during the same period were selected as the control group. The protein expressions of sCTLA-4 and RAD51C were compared between the two groups and patients with or without recurrence. Logistic regression was used to analyze the influencing factors of postoperative recurrence of cervical cancer patients, and a nomogram model of postoperative recurrence of cervical cancer patients was constructed and verified by calibration curve. The postoperative recurrence rate of cervical cancer patients with different sCTLA-4 and RAD51C protein expressions was compared.Results:The level of sCTLA-4 and the high expression rate of RAD51C protein in the study group were higher than those in the control group ( P<0.05). High-risk human papillomavirus positive, vascular infiltration, interstitial infiltration ≥1/2, paracterine infiltration, high expression of RAD51C protein and high SCTLA-4 level were independent risk factors for postoperative recurrence of cervical cancer ( P<0.05). High-risk human papillomavirus, vascular invasion, interstitial invasion, parametrial invasion, RAD51C protein and sCTLA-4 levels were used to construct a nomogram prediction model for postoperative recurrence of cervical cancer patients. The consistency indices were 0.610 (95% CI: 0.511-0.702), 0.616 (95% CI: 0.517-0.708), 0.640 (95% CI: 0.541-0.730), 0.609 (95% CI: 0.510-0.702), 0.728 (95% CI ranged from 0.633 to 0.809), 0.817 (95% CI ranged from 0.731 to 0.885), and the calibration curve validation showed high consistency. The net benefit rate of combined detection of sCTLA-4 and RAD51C proteins was greater than that of single detection. Conclusions:sCTLA-4 and RAD51C proteins are highly expressed in cervical cancer patients, and the high expression of both indicates that cervical cancer patients have a higher risk of recurrence after surgery. Clinically, the detection of sCTLA-4 and RAD51C protein expression can be used to screen patients with high recurrence risk.

OBJECTIVE To provide reference for rational use of cyclin-dependent kinase 4/6 （CDK4/6） inhibitors. METHODS Retrieved from Web of Science， PubMed， SpringerLink， CNKI， Wanfang Data and VIP database， and so on， the literature about lung toxicity related to CDK4/6 inhibitors were collected and analyzed statistically with Excel 2013 software. RESULTS A total of 12 literature which met the inclusion and exclusion criteria were included； 13 patients were involved， among which 3 cases were from the United States， 3 from Japan， 2 from India， and 1 from Israel， Spain， France， Australia and Saudi Arabia respectively； all patients were female， aged between 43-89 years， of whom 8 were treated with palbocicilib， 3 with abemacilib， and 2 with ribociclib. The lung toxicity of patients after medication occurred from 1 week to 15 months； the majority of patients were hospitalized with the symptom such as difficulty breathing， chest tightness， shortness of breath， dry cough， etc. The lung toxicity mainly manifested as interstitial lung disease， eosinophilic pneumonia， mediastinal and pulmonary granulomatous reaction， drug-induced pneumonia， diffuse alveolar damage， organizing pneumonia and so on. The shortest treatment duration was 3 weeks， and the longest was 6 months. The treatment measures included drug withdrawal， intravenous use of antibiotics， intravenous use of systemic steroids， oxygen inhalation， and so on； after treatment， 8 patients improved or recovered， and 5 patients died due to deterioration. One patient developed lung toxicity again after reuse of such drugs and must stop drugs permanently. CONCLUSIONS Lung toxicity related to CDK4/6 inhibitors possibly cause mortality. It is necessary to make early judgment， stop the drug in time， and give patients systemic steroids， oxygen inhalation and other treatment measures as soon as possible.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*

Humans ; Parkinson Disease/diagnosis* ; Machine Learning

Self-assembly is the basis of the formation of biological macromolecular structure. Enzyme-instructed self-assembly (EISA) with the help of tool enzymes, realizing the conversion of small molecular compounds to supramolecular nanostructures at specific sites, become a new strategy for drug discovery.In recent years, the exploration of EISA for developing malignant cancer therapy and imaging has made considerable progress, achieving the precise regulation and tumor targeting of nanostructures. This paper reviews the latest progress of EISA in the field of tumor diagnosis and treatment, the functions and characteristics of tool enzymes such as alkaline phosphatase, sirtuin, tyrosinase, γ-glutamyltranspeptidase and caspase-3,summarizes the research status of EISA targeting multiple organelles in tumor therapy, and introduces the application of EISA in tumor imaging, aiming to provide reference forthe research of EISA strategy in tumor diagnosis and treatment.