BACKGROUND:The implant osseointegration rate of patients with diabetes is low,and the failure rate is high,which seriously affects the quality of life.It is urgent to improve the implant osseointegration of patients with diabetes by effective means to elevate the success rate.Exploring the effect of berberine on the osteogenic differentiation of bone marrow mesenchymal stem cells under a high-glucose environment and its specific mechanism will provide effective theoretical support for solving the above problems. OBJECTIVE:To explore the effect of natural extract berberine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells under the high-glucose microenvironment. METHODS:Bone marrow mesenchymal stem cells of SD rats were cultured by the whole bone marrow adherence method.CCK-8 assay was used to detect the effects of different concentrations of berberine on the proliferation of bone marrow mesenchymal stem cells under the high-glucose environment and to screen out the optimal berberine concentration.The expressions of Runx2 and Osx were detected by alkaline phosphatase activity,alicarin red staining and PCR to determine the effect of berberine on osteogenic differentiation of bone marrow mesymal stem cells under the high-glucose environment.To further explore the underlying mechanism,we introduced the AMPK-specific inhibitor Dorsomorphin and used a DCFH-DA reactive oxygen species fluorescent probe to examine reactive oxygen species levels.The p-AMPK expression was also determined by western blot assay. RESULTS AND CONCLUSION:(1)10 μmol/L was the optimal concentration of berberine to promote bone marrow mesenchymal stem cell proliferation.(2)Alberberine promoted alkaline phosphatase viability of bone marrow mesenchymal stem cells and mineralized nodule formation in a high-glucose microenvironment.(3)Alberberine promoted the expression of Runx2 and OSx in a high-glucose microenvironment.(4)Alberensine effectively inhibited the reactive oxygen species level of bone marrow mesenchymal stem cells in a high-glucose environment.(5)The effects of berberine on promoting bone marrow mesenchymal stem cell osteogenesis and inhibition of reactive oxygen species were reversed by the AMPK inhibitor.(6)Berberine activated AMPK and promoted p-AMPK expression.(7)The above results indicate that berberine(10 μmol/L)promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment by activating AMPK and reducing intracellular reactive oxygen species levels.

Objective:To evaluate the relationship between gluten-free diet and rheumatoid arthritis (RA).Methods:Data were obtained from large-scale genome-wide association studies (GWAS), and genetic loci that are independent of gluten-free diet and RA of people of Europe2 were selected as instrumental variables. The gluten-free diet GWAS data included 64 949 individuals and 9 851 867 controls. Data were obtained from GWAS of 58 284 RA patients and 13 108 512 controls. The inverse variance weighted (IVW), MR Egger, weighted median method and weighted model were used to conduct two sample Mendelian randomization (MR) analysis. Cochran Q test and mendelian randomness pleiotropy residual sum and outlier (MR-PRESSO) were used to assess SNP heterogeneity. Applying the MR Egger intercept to test the level pleiotropy of SNP. The sensitivity analysis of the "leave one method" that evaluates whether MR studies were influenced by a single SNP. Results:After matching GFD and RA data, three SNPs were included as instrumental variables in the study. IVW showed that GFD could significantly reduce the risk of RA ( β=-60.83, s x=3.82, P<0.001). The weighted median method and weighted pattern also showed that the gluten free diet could reduce the risk of RA ( β=-57.97, s x=4.41, P<0.001; β=-55.81, s x=5.10, P=0.008). Sensitivity analysis of the correlation between GFD and RA showed that there might be heterogeneity between SNPs (Cochran Q test, Q=12.80, P=0.002). The MR-PRESSO results showed that no abnormal SNP was detected ( P=0.174). The forest map showed that SNPs was closely related to GFD and RA stability. The method comparison chart showed that the results of multiple testing methods were basically consistent. The funnel plot showed that SNPs were basically symmetrical, indicating that there was no pleiotropy in MR analysis. The MR Egger intercept test showed no horizontal pleiotropy in MR analysis (intercept value was-0.24, P=0.174). The sensitivity analysis of the "leave one method" is suggested that no single SNP had a significant impact on the overall results. Conclusion:Gluten free diet is related to the risk reduction of RA.

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.

[Objective]To evaluate the knowledge,attitude and practice(KAP)of medication use and safety among the patients with acquired immune deficiency syndrome(AIDS)in a 3A-grade hospital of Guangzhou city,and to provide scientific basis for AIDS prevention and treatment.[Methods]A questionnaire survey was conducted among AIDS patients aged 18 years and above in our hospital to investigate their KAP regarding medication use and safety.[Results]A total of 549 questionnaires were collected,of which 503 were valid,with an effective recovery rate of 91.6%.The average scores of KAP were(14.58±8.49),(25.21±6.92)and(47.58±3.33),respectively,with the scoring rates of 36.46%,63.02%,and 95.16%,respectively.There were statistically significant differences(P＜0.05)in knowledge scores among people with different ages,education levels and occupations.Multiple linear regression showed that education level and medical insurance status had most significant impact on knowledge scores(P<0.05).Significant differences were found in attitude scores among people with different education levels(P<0.05),as well as in practice scores among people with different occupations(P<0.05).Multiple linear regression revealed that age,occupation,knowledge score and attitude score had a significant impact on practice scores(P<0.05).Patients expected to receive pharmaceutical care services from the pharmacists via face-to-face communication,network platform and telephone consultation on medication knowledge such as adverse drug reactions and response measures,drug-drug interactions,missed medication and response measures,medication adherence measures,etc.[Conclusions]AIDS patients in this hospital have a good awareness of medication safety,but their knowledge of medication use needs improvement.Some bad habits may affect their compliance,resulting in safety hazards.Therefore,there is an urgent demand for pharmaceutical care services related to rational drug use.

Objective:To explore the application effects of case nursing review based on training transfer theory in specialized nurses training, and to provide reference for promoting the knowledge and skills transformation of specialized nurses.Methods:A total of 64 specialized nurses from Longgang District Maternity & Child Healthcare Hospital of Shenzhen City were selected as subjects by convenient sampling from June 10 to July 20, 2022 and divided into control group and observation group by random number table method. The control group received routine core competence training, while the observation group received case nursing review based on training transfer theory. The training duration of both groups was 12 months. Within 1 month after the training, the training satisfaction, nursing case report quality score, core competence of specialized nurses, patient satisfaction and training transfer outcomes of the two groups were compared.Results:Finally, 62 specialist nurses were included, with 31 nurses in control group and observation group, 2 males and 29 females in each group, aged (34.10 ± 3.93) years in control group and (33.35 ± 3.06) years in observation group. Before training, there were no significant differences in nursing case report quality score, total score of core competence of specialized nurses and patient satisfaction between the two groups (all P> 0.05). After training, the nursing case quality score, total score of core competence and patient satisfaction in the observation group were 73.00(68.00, 78.00), 179.00(175.00, 183.00), 96.00(94.00, 99.00) points, which were higher than 85.00(80.00, 89.00), 163.00(162.00, 166.00), 88.00(85.00, 92.00) points in the control group, and the differences were statistically significant ( Z = 5.81, 6.73, 13.31, all P<0.05). The total training satisfaction of nurses, awards in professional competitions and the number of technological innovations in the observation group were 21.00(20.00, 22.00) points, 15 awards, 8 innovations, which were higher than 15.00(14.00, 16.00) points, 5 awards, 2 innovations in the control group, and the differences were statistically significant ( Z = 6.74, χ2 = 7.38, 4.29, all P<0.05). Conclusions:The application of case nursing review based on training transfer theory can improve the training satisfaction of specialist nurses, stimulate their motivation to transfer training, and improve their ability and effectiveness in training transfer.

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 investigate effects of lentinan(LNT)on atherosclerosis(AS)plaque formation and adenosine 5'-mo-nophosphate-activated protein kinase(AMPK)signaling pathway in ApoE-/-mice.Methods:ApoE-/-mice were divided into model group,Lipitor group(5 mg/kg),LNT low(5 mg/kg),medium(10 mg/kg),high(20 mg/kg)dose groups,C57BL/6 mice of similar genetic background were as control group.After 12 weeks of intragastric administration,automatic biochemical analyzer was used to detect serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein(LDL-C)and high-density lipoprotein(HDL-C)levels;serum levels of IL-6 and IL-1β were detected by ELISA;oil red O staining was used to observe overall aortic plaque formation;HE staining was used to observe formation of plaque in aortic roots;AMPK pathway proteins expressions in aortawas were detected by Western blot.Results:Compared with control group,serum TC,TG,LDL-C,IL-6,IL-1β levels,aortic plaque area,aortic root plaque area,aortic tissue NLRP3,IL-6,IL-1β expressions were significantly increased in model group,p-AMPK/AMPK expression in aortic tissue and serum HDL-C level were reduced significantly(P<0.05);compared with model group,TC,TG,LDL-C,IL-6,IL-1β,aortic plaque area,aortic root plaque area,aortic tissues NLRP3,IL-6 and IL-1β expressions were reduced significantly in Lipitor group and medium,high dose LNT groups,p-AMPK/AMPK expression in aortic tissue and serum HDL-C level were increased significantly,difference between each dose of LNT was statistically significant(P<0.05);there was no significant difference between Lipitor group and LNT high-dose group(P>0.05).Conclusion:LNT may inhibit inflammatory response and formation of AS plaques in AS mouse by activating AMPK pathway.

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

Objective To investigate the effect of intestinal mucosal Toll-like receptor 4/nuclear factor κB (TLR4/NF-κB) signaling pathway on renal damage in pseudo-sterile IgA nephropathy (IgAN) mice. Methods C57BL/6 mice were randomly divided into experimental group (pseudosterile mouse model group), control group (IgAN mouse model group), pseudosterile mouse blank group, and normal mouse blank group. Pseudosterile mice were established by intragastric administration of quadruple antibiotics once a day for 14 days. The pseudosterile IgAN mouse model was set up by combination of oral bovine serum albumin (BSA) administration and staphylococcal enterotoxin B (SEB) injection. The pathological changes of renal tissue were observed by immunofluorescence staining and PAS staining, and the intestinal mucosa barrier damage indicators lipopolysaccharide(LPS), soluble intercellular adhesion molecule 1(sICAM-1) and D-lactate(D-LAC) were analyzed by ELISA. Biochemical analysis was used to test 24 hour urine protein, serum creatinine and blood urea nitrogen. The mRNA and protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor κB (NF-κB) were detected by reverse transcription PCR and Western blot analysis. Results The kidney damage of pseudosterile IgAN mice was more severe than that of IgAN mice, and the expressions of intestinal mucosal barrier damage markers (LPS, sICAM-1 and D-LAC) were significantly increased in pseudosterile IgAN mice. In addition, the expressions of TLR4, MyD88, and NF-κB level were all up-regulated in the intestinal tissues of IgAN pseudosterile mice. Conclusion Intestinal flora disturbance leads to intestinal mucosal barrier damage and induces activation of TLR4 signaling pathway to mediate renal injury in IgAN.
Animals ; Mice ; Mice, Inbred C57BL ; Glomerulonephritis, IGA ; NF-kappa B ; Toll-Like Receptor 4/genetics* ; Lipopolysaccharides ; Myeloid Differentiation Factor 88/genetics* ; Kidney ; Intestinal Mucosa ; Infertility ; Disease Models, Animal