Objective To investigate and analyze clinical characteristics and related factors of elderly patients with type 2 diabetes mellitus (T2DM) and frailty. Methods A total of 310 elderly patients with T2DM admitted to the hospital from January 2023 to June 2024 were selected as the research subjects. Their general information and disease-related information was collected through questionnaires. The Fried Frailty Scale was used to evaluate frailty status, and the patients were divided into frailty group and non-frailty group based on the Fried Frailty Scale score. Factors related to T2DM with frailty in the elderly were analyzed. Results The incidence of frailty in this study was 31.61% (98/310), and those without frailty accounted for 68.39% (212/310). There were statistically significant differences between the two groups in terms of age, body mass index (BMI), Self-rating Depression Scale (SDS) score, number of chronic complications, glycosylated hemoglobin (HbA1c) level, hemoglobin level, Mini-Nutritional Assessment-Short Form (MNA-SF) score, and Charlson Comorbidity Index (CCI) score (P<0.05). Multivariate logistic regression analysis showed that age, HbA1c level, SDS score, MNA-SF score, and CCI score were risk factors for frailty in elderly patients with T2DM (P<0.05). Conclusion The incidence of frailty is relatively high in elderly patients with T2DM. It is influenced by factors such as age , SDS score , HbA1c level , MNA-SF score and CCI score, and deserves clinical attention.

Objective: To explore the damaging effect of the IP3-protein kinase C pathway on human retinal vascular endothelial cells under high glucose conditions. Methods: Human retinal vascular endothelial cells in the logarithmic growth phase were taken and divided into the experimental group(10 mmol/L glucose and 20 mmol/L glucose) and the control group(5 mmol/L glucose). The morphological changes of cells were observed under different concentrations of glucose culture medium. The apoptosis of human retinal vascular endothelial cells was detected by flow cytometry. Real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to detect the RNA expression level of PKC in the IP3-protein kinase C(PKC) pathway, and Western blot was used to detect the protein expression level of PKC in cells. Results: Under high glucose conditions, the volume of human retinal vascular endothelial cells increased, the degree of extension decreased, and the apoptosis rate increased. Compared with the control group, the apoptosis rate of human retinal vascular endothelial cells in the 20 mmol/L glucose group increased(P<0.05); the IP3 level of human retinal vascular endothelial cells in the 20 mmol/L glucose group(587.9±15.2) ng/ml was lower than that of the control group(738.9±1.0) ng/ml(P<0.05). Under high glucose treatment, the expression levels of PKC mRNA and protein increased(P<0.05). Conclusion The IP3-PKC pathway may be involved in the damage process of human retinal vascular endothelial cells under high glucose conditions, and thus may play a role in diabetic retinopathy.

Objective To investigate the effectiveness of team-based learning(TBL)combined with research-based learning(RBL)in enhancing evidence-based nursing practice skills of undergraduate nursing interns.Methods A total of 114 undergraduate nursing students who interned in the operating room of a ⅢA hospital in Guangzhou from July 2021 to April 2022 were selected as study subjects.A randomized cluster sampling method based on a random number table was used to divide the students into a control group and a trial group.The control group received traditional teaching methods,while the trial group was taught using a combination of TBL and RBL.The two groups were compared in terms of evidence-based practice skills,critical thinking abilities,and their evaluations on the teaching methods.Results The differences in evidence-based practice skills and critical thinking abilities before and after the internship were significantly higher in the observation group compared to the control group(t=35.108,35.897;both P<0.05).Additionally,post-internship evaluation scores for the teaching methods in the trial group were significantly higher than those in the control group(t=-17.580,P<0.05).Conclusion TBL combined with RBL effectively enhances the evidence-based nursing practice skills and critical thinking abilities of undergraduate nursing interns.This approach also improves students'evaluations on the teaching methods and fosters the cultivation of excellent clinical evidence-based nursing professionals.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore the relationship between the neural development of preterm infants and gut microbiota.Methods:66 premature infants who were hospitalized in the Neonatology Department of Hunan Children′s Hospital from September 2018 to September 2019 were included in the study. Their fecal samples and clinical data from the first admission were collected. According to the neurodevelopment, the patients were divided into normal neurodevelopment group and neurodysplasia group. The bacterial DNA of fecal samples was extracted by 16S rDNA high-throughput sequencing technology and bioinformatics analysis was conducted to compare the composition and diversity of gut microbiota between the two groups.Results:(1) The Shannon index of gut microbiota in normal neurodevelopmental group and neurodysplastic group was 0.89(0.41, 1.51) and 1.01(0.47, 1.31), respectively. There was no significant difference in diversity index between the two groups ( P>0.05). (2) Bifidobacterium, veronica and negativites in the gut microbiota of the normal neurodevelopmental group were significantly higher (all P<0.05), and streptococcus in the gut microbiota of the dysplastic group were significantly higher ( P<0.05). The gut microbiota of the two groups were mainly enterococcus and escherichia shigella. Conclusions:At the genus level, enterococcus and escherichia are the dominant flora of early gut microbiota in preterm infants. Gut microbiota is related to the neural development of preterm infants. The increased abundance of streptococcus, and the decreased abundance of bifidobacterium, veronicus, and negativites may be risk factors for neurodysplasia of preterm infants. The diversity of gut microbiota in early preterm infants may not be significantly related to neural development.

Objective:To compare the characteristics and differences of intestinal flora in premature infants with late-onset sepsis (LOS) and pneumonia by high-throughput sequencing technology, and to investigate the relationship between intestinal flora and LOS.Methods:Through the case-control method, premature infants with late-onset sepsis who were hospitalized in the neonatal department of Hunan Children′s Hospital from August 2018 to October 2019 were selected as the case group ( n=8). At the same time, premature infants diagnosed with pneumonia were selected as the control group ( n=8). The fecal samples of 16 premature infants were collected for the first time, and the DNA was extracted. The DNA of the target region was amplified by polymerase chain reaction(PCR). High-throughput sequencing was performed using NovaSeq 6000 platform to analyze the composition and diversity of intestinal flora between the two groups. Results:(1) Alpha diversity analysis showed that there was no significant difference in the richness and diversity of intestinal flora between the two groups(all P>0.05). (2) The intestinal flora in premature infants of LOS group and control group were dominated by Firmicutes and Proteobacteria, and facultative anaerobes such as Enterococcus and Escherichia-Shigella were the dominant flora at the genus level. Metastas statistical analysis showed that there was no statistically significant difference in flora composition between the two groups at the phylum level ( P>0.05). (3) Metastas statistical analysis was carried out at the level of class, order, family, genus, and species. The relative abundance of actinomycetes, digestive streptococcaceae and Clostridium in LOS group was higher than that in pneumonia group, and the difference was statistically significant (all P<0.05). (4) The relative abundance of Staphylococcus in the LOS group was significantly greater than that in the control group, but Metastas statistical analysis showed that there was no statistically significant difference in the relative abundance of staphylococcus between the two groups ( P>0.05). (5) Among the 8 cases of LOS, 3 premature infants had positive blood cultures, namely Streptococcus agalactiae, Streptococcus mitis, and Enterococcus faecalis. Enterococcus faecalis belongs to the genus Enterococcus, and Enterococcus belongs to the dominant genus in the LOS group. Conclusions:Different site infections have effects on intestinal microecology of premature infants. There were differences in intestinal flora composition between premature infants with LOS and premature infants with pneumonia.

The recombinant adenoviruses expressing miR-22 (Ad-miR-22) was constructed and the effect of Ad-miR-22 on insulin signal pathway and glucose uptake in HepG2 cells was analyzed. MiR-22 gene was amplified by PCR from human hepatocytes and cloned into the pAdTrack-CMV vector to generate the shuttle plasmid pAdT-22. The positive colonies were confirmed by PCR and sequencing. The resultant shuttle plasmid was linearized with Pme I, followed by co-transformation into competent BJ5183 cells containing an adenoviral backbone plasmid (pAdEasy-1) to create the recombinant plasmid pAd-miR-22. After digested with Pac I, the linearized pAd-miR-22 was transfected into 293A packaging cell line to generate recombinant adenoviruses Ad-miR-22. HepG2 cells were infected with Ad-miR-22 or control Ad-GFP (adenoviruses expressing green fluorescent protein), and then the miR-22 expression levels were analyzed by qPCR. The result shows that adenovirus-mediated overexpression of miR-22 significantly decreased insulin-induced glucose uptake in HepG2 cells. Moreover, overexpression of miR-22 markedly decreased insulin-induced phosphorylation of GSK-3β. miR-22 also increased the mRNA levels of gluconeogenic genes in HepG2 cells. Furthermore, Western blotting results indicate that the protein expression of SIRT1 decreased in Ad-miR-22 infected HepG2 cells as compared with Ad-GFP infected HepG2 cells. In summary, overexpressing of miR-22 significantly increased gluconeogenesis while decreased glucose uptake in HepG2 cells. The effect of miR-22 on glucose metabolism may be mediated by SIRT1.
Adenoviridae ; genetics ; Glucose ; metabolism ; Glycogen Synthase Kinase 3 beta ; metabolism ; Hep G2 Cells ; Humans ; MicroRNAs ; genetics ; metabolism ; Signal Transduction ; genetics ; Transfection

The discovery of hydroxylases in the anticancer drug taxol biosynthesis pathway is a hotspot and difficulty in current research. In this study, a new hydroxylase gene TcCYP725A22 (GenBank accession number: MF448646.1) was used to construct a sub-cellular localization vector pCAMIBA1303-TcCYP725A22-EGFP to get the transient expression in onion epidermal cells. Laser confocal microscopy revealed that the protein encoded by this gene was localized in the cell membrane. Furthermore, the recombinant plant expression plasmid pBI121-TcCYP725A22 was constructed. After transient transformation to the Taxus chinensis mediated by Agrobacterium tumefaciens LBA4404, qRT-PCR and LC-MS were utilized to analyze the effects of TcCYP725A22 overexpression on the synthesis of taxol. The results showed that, in the TcCYP725A22 overexpressed cell line, expression levels of most defined hydroxylase genes for taxol biosynthesis were increased, and the yield of taxanes were also increased. It was concluded that the hydroxylase gene TcCYP725A22 is likely involved in the biosynthetic pathway of taxol.
Biosynthetic Pathways ; Mixed Function Oxygenases ; Paclitaxel ; Taxoids ; Taxus

Objective To explore the clinical features,genetic causes and prognosis of intellectual disability with epilepsy(ID-E)in children.Methods The data of unknown causes of ID-E children(n=40)who were identified in Department of Pediatrics,Xiangya Hospital of Central South University from March 2015 to March 2016 were respectively analyzed,and follow-up studies were performed to investigate the epilepsy control and intellectual deve-lopment.Results Forty unexplained ID-E included 25(62.5％)male,and 34(85.0％)cases were severe intellectual disability patients.The onset age of epilepsy was 0.16 to 8.00 years old,median age was 1.5 years old.Twenty cases(50.0％)had slow electroencephalogram background,and 22 cases(55.0％)had focal spikes.Ten cases(25.0％)had abnormal cranial images,with brain dysplasia or atrophy.Follow-up lasted from 0.58 to 1.58 years,and 19 cases(47.5％)had seizure control.Twenty-five cases(62.5％)had used at least 2 anti-epilepsy drugs during follow-up,and 19 cases(47.5％)had drug refractory epilepsy.Improvement of mental or motor development in epilepsy controlled group and the uncontrolled group were 12 cases(63.2％)and 2 cases(9.5％).There were separately 8 cases(8/40 cases,20.0％)and 3 cases(3/16 cases,18.8％)diagnosed respectively by whole genome-wide analysis of copy number variants(CNVs)and gene-panel whose CNVs test findings were negative.Conclusions ID-E patients of unknown causes have the following clinical features:they were mostly found in male patients with severe intellectual disability,and drug refractory epilepsy patients have rather high percentage;well controlling of epilepsy is useful for improvement of mental and motor development.Genetic analysis is significant for control and prognosis of ID-E patients,and genome-wide CNVs have high positive rates which can be used as first-tier test to detect genetic etiology of ID-E of unknown cause.

Objective To explore the febrile response and placental pathological inflammation of pregnant rats exposed to intrauterine infection in late gestation.Methods Pregnant Sprague-Dawley rats at gestational day 18 were randomly divided into control group and intrauterine-infected group with six rats in each.The intrauterine-infected group was intraperitoneally injected with 350 μ g/kg lipopolysaccharide to establish a rat model of intrauterine infection,while the control group was injected with sterile saline of the same dose.Core temperature was measured every 1 h after intraperitoneal injection of lipopolysaccharide or saline for 8 h.At gestational day 19,after anesthesia,the placentas were taken and stained with HE.The expression levels of tumor necrosis factor-α,interleukin-6,and interleukin-1β in the placenta were determined by enzyme linked immunosorbent assay.Student t test was used for statistical analysis.Results (1) There was no temperature difference between the two groups before experimental treatment (P ＞ 0.05).Core temperature was increased 1 h after the lipopolysaccharide injection,reaching (37.67 ±0.08) ℃.The increase of temperature was significant compared with the control group [(37.13 ± 0.08) ℃,t=10.178,P ＜ 0.01].Fever was lowered 2 h later and the rats became hypothermic with body temperature below 37 ℃ in the intrauterine-infected group.The body temperature in the intrauterine-infected group after 2-6 h was (37.70 ± 0.10),(37.23 ± 0.05),(36.57 ± 0.06),(36.60 ± 0.10) and (36.57 ± 0.08) ℃,respectively,compared with the control group [(36.83 ±0.12),(36.63 ± 0.12),(36.71 ± 0.07),(36.87±0.12),and (36.77±0.08) ℃,respectively],the differences being all statistically significant (t=11.402,11.163,-4.025,-4.000 and-4.243,all P ＜ 0.01).(2) HE staining revealed large amounts of neutrophils infiltration,vascular enlargement and congestion in the placenta of the intrauterine-infected rats.No inflammatory cell infiltration was observed in the control placentas.(3) The expression levels of proinflammatory cytokine tumor necrosis factor-α [(0.62 ± 0.02) ng/g],interleukin-6 [(66.12 ± 5.11) ng/g],and interleukin-1β [(7.09± 1.23) ng/g] in the intrauterine-infected group were higher than those in the control group [(0.27±0.01),(16.71 ±1.55) and (2.86 ± 0.38) ng/g,respectively].The differences were all statistically significant (t=-26.608,-18.749 and-5.714,all P ＜ 0.01).Conclusion After exposure to lipopolysaccharide in late gestation,pregnant rats show significant inflammatory response in the placenta,with suppression of febrile response and presence of hypothermia.