Objective To construct plasmids and knock out HIF-1α gene expression in an naked mole rat skin fibroblasts(NSF)cell line using CRISPR/Cas9 genomic editing technology,to provide an in vitro cell model for studying the mechanism of hypoxia tolerance and the occurrence and development of hypoxia-related diseases in naked mole rats.Methods We designed four pairs of single guide RNA(sgRNA)sequences targeting exons 1～4 of the NSF HIF-1αgene and successfully constructed an expression plasmid.The plasmid with the optimal sgRNA was identified and transfected into 293T cells,and the supernatant was used for detecting the virus titer.Lentivirus particles carrying sgRNAs of HIF-1α were transfected into NSF cells which express Cas9 protein,based on a previous protocol.After transfection,fluorescence signals were observed under a fluorescence microscope,and HIF-1α expression in NSF cells was detected by Western Blot and T7 endonuclease 1(T7E1)analysis.Results Sanger sequencing showed that the designed sgRNA was successfully inserted into pX459 and pKLV2-U6-sgRNA2 vectors,demonstrating successful construction of a recombinant plasmid for transfection.T7E1 digestion successfully removed three bands and the target efficiency of sgRNA was 54％.Western Blot showed that the HIF-1α gene was successfully knocked out and its protein level was significantly reduced in NSF cells from naked mole rats(P=0.0019).There were no obvious morphological changes in HIF-1α-knockout cells under the microscope,and gene knockout had no obvious effect on cell proliferation.Conclusions We successfully constructed an HIF-1α-knockout cell line using CRISPR/Cas9 technology,to provide an experimental basis for further studies of the biological function of HIF-1α,as well as the mechanism of hypoxia tolerance in naked mole rats.The result also provide a theoretical foundation for the prevention and treatment of hypoxia-related diseases.

Objective:To compare the mechanical properties between our self-designed axially controlled compression spinal rod (ACCSR) and conventional spinal rod (CSR) for lumbar spondylolysis (LS).Methods:This study selected 36 ACCSRs (the ACCSR group) and 36 CSRs (the CSR group), both of which were in a diameter of 6.0 mm and manufactured in the same batch. They were subjected respectively to biomechanical tests of spinal rod and pedicle screw-rod internal fixation system. In spinal rod tests: the stiffness and yield load of the spinal rods were calculated using four-point bending tests ( n=7) and comparisons were made between the 2 groups; spinal rod fatigue tests ( n=8) recorded the successful compression loads after 2.5 million cycles of loading and compared them with the maximum force at the isthmus of a normal adult's unilateral lumbar spine (198.72 N). In tests of the pedicle screw-rod internal fixation system, the axial compression tests ( n=7) measured the axial gripping capacity, the axial torsion tests ( n=7) the torsional gripping capacity, and the lateral compression tests ( n=7) the stiffness and yield load of pedicle screws in the 2 groups respectively. Results:The stiffness [(1,543.37±61.41) N/mm] and yield load [1,338.57 (1,282.00, 1,353.80) N] of ACCSR group were significantly smaller than those of CSR group [(3,797.63±156.15) N/mm and 4,059.95 (3,813.80, 4,090.89) N] ( P<0.05). The spinal rod fatigue tests showed that the respective loads of CSR and ACCSR passing the 2.5 million fatigue tests were 640.00 N and 320.00 N, both larger than the maximum force at the unilateral lumbar isthmus of a normal adult (198.72 N). There were no significant differences between the ACCSR group and the CSR group in the axial gripping capacity and torsional gripping capacity, as well as in stiffness and yield load of screws between the 2 groups ( P>0.05). Conclusions:In fixation of LS, although the yield load, stiffness and fatigue resistance of ACCSR are inferior to those of CSR, the biomechanical properties of the two sets of pedicle screw-rod internal fixation system are comparable. The fatigue resistance of ACCSR can meet the stress requirements of the normal human isthmus.

Tissue engineering bone technology, grounded in seed cells, cytokines, and scaffold supports, provides an effective solution for addressing extensive bone defects, demonstrating significant potentials in the field of bone repair. However, this technology still faces numerous challenges. Focusing on vascularization in engineered bones, this article reviews various methods to enhance vascularization within tissue-engineered bones, including multicellular co-culture, application of angiogenic factors, advanced 3D printing, and aid of surgical interventions. This article also analyses the latest research developments and the limitations of the methods, and speculates future research directions for tissue engineered bone.

BACKGROUND: Colorectal carcinogenesis and progression are related to the gut microbiota and the tumor immune microenvironment. Our previous clinical trial demonstrated that berberine (BBR) hydrochloride might reduce the recurrence and canceration of colorectal adenoma (CRA). The present study aimed to further explore the mechanism of BBR in preventing colorectal cancer (CRC). METHODS: We performed metagenomics sequencing on fecal specimens obtained from the BBR intervention trial, and the differential bacteria before and after medication were validated using quantitative polymerase chain reaction. We further performed ApcMin/+ animal intervention tests, RNA sequencing, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: The abundance of fecal Veillonella parvula ( V . parvula ) decreased significantly after BBR administration ( P = 0.0016) and increased through the development from CRA to CRC. Patients with CRC with a higher V. parvula abundance had worse tumor staging and a higher lymph node metastasis rate. The intestinal immune pathway of Immunoglobulin A production was activated, and the expression of TNFSF13B (Tumor necrosis factor superfamily 13b, encoding B lymphocyte stimulator [BLyS]), the representative gene of this pathway, and the genes encoding its receptors (interleukin-10 and transforming growth factor beta) were significantly upregulated. Animal experiments revealed that V. parvula promoted colorectal carcinogenesis and increased BLyS levels, while BBR reversed this effect. CONCLUSION: BBR might inhibit V. parvula and further weaken the immunomodulatory effect of B cells induced by V. parvula , thereby blocking the development of colorectal tumors. TRIAL REGISTRAION ClinicalTrials.gov, No. NCT02226185.
Animals ; Humans ; Berberine/therapeutic use* ; Carcinogenesis ; Veillonella ; Colorectal Neoplasms/genetics* ; Tumor Microenvironment

Objective:To retrieve, evaluate and summarize the relevant evidence on the management of hyperphosphatemia in adult maintenance hemodialysis patients.Methods:Domestic and foreign databases and relevant professional websites were searched for best practices, evidence summaries, guidelines, expert consensus and systematic reviews on hyperphosphatemia in adult maintenance hemodialysis patients. The retrieval time was from the establishment of the databases to January 2023. The methodological quality of the included articles was evaluated, and the best evidence was extracted and summarized.Results:A total of 13 articles were included, including 2 clinical decisions, 4 guidelines and 7 systematic reviews. A total of 23 evidences were summarized from 5 aspects, including blood phosphorus monitoring, adequate dialysis, diet control, drug management and health education.Conclusions:This study summarizes the best evidence on the management of hyperphosphatemia in maintenance hemodialysis patients and provides a reference for clinicians to manage hyperphosphatemia in adult maintenance hemodialysis patients.

Objective To use the "three-dimensional quality structure" model as a theoretical basis, based on multiple evidence-based evidence, to construct ICU nursing sensitivity quality indicators, and to provide reference for the evaluation and monitoring of ICU department nursing safety quality. Methods Evidence-based and Delphi methods were used to complete 3 rounds of inquiry among 27 experts to obtain consensus opinions of experts and to determine nursing quality indicators. Results Of the 30 articles included, 17 wereⅢA, 10 wereⅣA, 2 wereⅤA, and 1 wasⅤB. According to the"three-dimensional structure theory"model, the included indicators were sorted out and a total of 12 items were obtained. The effective recovery rate of 3 rounds of questionnaires was 96.6％, 100.0％, and 96.4％. The authoritative coefficients were 0.899, 0.895, and 0.909, and the coordination coefficients of the first, second, and third indexes were 0.340, 0.379, and 0.403, respectively,with significance (P<0.01). The final ICU nursing sensitivity quality indicator consists of 3 first-level indicators, 15 second-level indicators and 69 third-level indicators. The selection of indicators at all levels gradually converges. Conclusions The established ICU nursing sensitivity quality index accords with the principle of scientificity and practicality, and can be used for ICU department nursing quality assessment.

Objective To evaluate the role of phosphatidylinositol 3-kinase ( PI3K)∕protein kinase B ( Akt) signaling pathway in propofol-induced reduction of intestinal ischemia-reperfusion ( I∕R) injury in rats. Methods Thirty-two healthy male Sprague-Dawley rats, aged 2-3 months, weighing 225-275 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group ( Sham group), intestinal I∕R group ( I∕R group), propofol group ( P group), and PI3K inhibitor wortmannin plus propofol group ( W+P group) . Intestinal ischemia was induced by occluding the superior mesenteric ar-tery for 45 min followed by 2 h of reperfusion to establish the model of intestinal I∕R injury. Propofol was in-travenously infused at a rate of 20 mg·kg-1 ·h-1 starting from the onset of reperfusion until the end of reper-fusion in group P. Wortmannin 15 μg∕kg was intravenously injected at 25 min before reperfusion, and propofol was intravenously infused at a rate of 20 mg·kg-1 ·h-1 starting from the onset of reperfusion until the end of reperfusion in group W+P. Rats were sacrificed at 2 h of reperfusion, and small intestinal tissues were obtained for microscopic examination of pathologic changes of intestinal mucosa and for determination of wet∕dry weight ratio (W∕D ratio), malondialdehyde (MDA) content (by thiobarbituric acid colorimetric method) , superoxide dismutase ( SOD ) activity ( using xanthine oxidase method ) , myeloperoxidase ( MPO) activity ( by MPO assay) , and phosphorylated Akt ( p-Akt) expression ( by Western blot) . Intes-tinal damage was assessed and scored according to Chiu. Results Compared with group Sham, Chiu' s score, W∕D ratio, MDA content and MPO activity were significantly increased, the SOD activity was de-creased, and p-Akt expression was down-regulated in group I∕R (P<0. 05). Compared with group I∕R, Chiu's score, W∕D ratio, MDA content and MPO activity were significantly decreased, the SOD activity was increased, and p-Akt expression was up-regulated in group P (P<0. 05). Compared with group P, Chiu's score, W∕D ratio, MDA content and MPO activity were significantly increased, the SOD activity was decreased, and p-Akt expression was down-regulated in group W+P (P<0. 05). Conclusion The mechanism by which propofol reduces intestrnal I∕R injury is related to activating PI3K∕Akt signaling path-way and inhibiting inflammatory and oxidative stress responses in rats.

Objective:To explore the wild medicinal plant species and utilization of Hasi mountain nature reserve,in order to provide the references for reasonable utilization and protection of the medicinal plant resources in this area. Method:The survey was conducted based on the technical scheme of the Fourth National Survey on Chinese Material Medica Resources. By consulting literature,collecting medicinal plants specimen and visiting survey area,researchers collected and summarized the wild medicinal plant species, and analyzed the results of reserves. Result:The results showed 7 varieties in 247 kinds of wild medicinal plants in Hasi mountain nature reserve,which belonged to 161 genera in 61 families;by the medicinal parts of all wild medicinal plants,the whole plant of a total of 124 species could be used,accounting for 48.82% of the total species,by traditional Chinese medicine efficacy, antipyretic herbs were the majority,accounting for 32.68% of the total. In addition,there were 8 kinds of rare and endangered wild medicinal plants,such as Ephedra sinica,Gentiana dahurica and Epipactis helleborine. Conclusion:Hasi mountain nature reserve has rich wild medicinal plant resources,and is an important part of medicinal plant resources and protection areas of Jingyuan county and Loess Plateau. However, because they are serious affected by incontinent excavation and insect pest,efforts shall be made in scientific protection and rational development.

Objective: To discuss the benefit of using transmastoid slotted labyrinthotomy approach (TSLA) and customized electrode for common cavity deformity (CCD) patients, and to evaluate the audiological outcomes. Methods: A retrospective analysis of the intraoperative monitoring data and postoperative auditory outcomes of 10 CCD cases who received cochlear implantation in Beijing Tongren Hospital,Capital Medical University from April 2016 to December 2017, was conducted using TSLA and customized electrod as a test group.At the same time, 10 cases of age and gender matched children with severe or severe sensorineural hearing loss and normal inner ear structures were recorded as a control group. Four questionnaires were collected from the two groupspre-operative, 6 months and 1 year after start-up. The Wilcoxon signed rank sum test was used to compare the scores of different time points. Rank sum test of two independent samples was used to compare the scores between different groups. Results: None of the patients exhibited CSF leakage or facial paralysis after operation. The average impedance of 8 CCD children in the experimental group was below 5 kΩ. The total extraction rate of auditory neural response telemetry (ART) of all stimulating electrodes was 55% (33/60), and the amplitude of the electricity induced complex action potential (ECAP) of electrodes at different positions ranged from 50.69 to 170.3 μV.The average surgical time of the TSLA group was 46.4 min, shorter than the traditional approach.There was significant difference between the scores of pre-operative, 6 months and 1 year after start-up for the TSLA group(categories of auditory performance,CAP: 2.0 (2.0,3.0) vs. 4.0 (3.8,4.0) , Z=-3.109, P=0.002; speech intelligibility rating,SIR: 2.0 (2.0,2.3) vs. 3.0 (2.8,4.0) , Z=-2.952, P=0.003; meaningful use of speech scale,MUSS: 4.0 (3.3,6.0) vs. 9.0 (6.0,11.8) , Z=-3.421, P=0.001; meaningful auditory integration scale, MAIS or infant-toddler and meaningful auditory integration scale, IT-MAIS: 5.5 (3.8,9.0) vs. 15.5 (10.8,18.5) , Z=-3.522, P=0.000 for the latter two).In addition, The scores of the TSLA group were significantly worse than the control group at 6 months and 1 year after start-up.The high-resolution CT scan showed good adhesion of the electrodes without displacementone week and one year after surgery. Conclusions For CCD patients, TSLA and customized electrode is recommended due to lower surgical difficulty and post-operative risk, shortened surgical time; Intraoperative ART monitoring can be used to determine the integrity of the auditory pathway. Children with common cavity deformity have a longer period of auditory rehabilitation, and the individual differences in speech rehabilitation are significantly different, which is significantly worse than those with normal structure.

Background:The incidence of gastric cancer is gradually rising in recent years,long non-coding RNA taurine up-regulated gene 1 (TUG1)may have certain effects on the occurrence and progression of gastric cancer. Aims:To study the expression TUG1 in gastric cancer tissue and its effect on prognosis of patients with gastric cancer,and study the correlation between TUG1 and p27,cyclin D1. Methods:Surgically resected gastric cancer tissues and corresponding distal normal tissues of 48 gastric cancer patients from June 2013 to December 2013 at Qingdao Municipal Hospital were collected. qRT-PCR was used to detect the mRNA expression of TUG1,and its relationship with clinicopathological features was analyzed. Protein expressions of p27 and cyclin D1 were determined by Western blotting,and correlation with expression of TUG1 was analyzed. Kaplan-Meier was used to analyze the relationship between expression of TUG1 and prognosis. Results:The mRNA expression of TUG1 in gastric cancer tissues was significantly higher than that in corresponding normal tissues (6. 18 ± 0. 19 vs. 5. 09 ± 0. 16,P < 0. 05),and was not correlated with gender,age,tumor size,but correlated with lymph node metastasis,tumor differentiation and TNM staging (P < 0. 01). The protein expression of p27 was significantly decreased in gastric cancer tissues than in normal tissues (0. 1709 ± 0. 0212 vs. 0. 3087 ± 0. 0252,P < 0. 01),while protein expression of cyclin D1 was significantly increased (0. 3417 ± 0. 0271 vs. 0. 2417 ± 0. 0173,P < 0. 01),and the expression of p27 was negatively correlated with expression of cyclin D1 in gastric cancer tissues (r = - 0. 897,P < 0. 01). The expression of TUG1 was negatively correlated with expression of p27 (r = - 0. 730,P < 0. 01),and was positively correlated with expression of cyclin D1 (r = 0. 809,P < 0. 01)in gastric cancer tissues. The median survival time was shorter in gastric cancer patients with high-expressed TUG1 than in patients with low-expressed TUG1 (P < 0. 05). Conclusions:Long non-coding RNA TUG1 plays a role of cancer gene in the development of gastric cancer through p27 /cyclin D1 pathway. Detection of expression of TUG1 has important significance on the prediction of prognosis of gastric cancer patients.