BACKGROUND:In recent years,numerous studies have shown that autophagy and mitophagy play an important role in the regulation of bone metabolism.Under non-physiological conditions,mitophagy breaks the balance of bone metabolism and triggers metabolism disorders,which affect osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells,etc. OBJECTIVE:To summarize the mechanism of mitophagy in regulating bone metabolic diseases and its application in clinical treatment. METHODS:PubMed,Web of Science,CNKI,WanFang and VIP databases were searched by computer using the keywords of"mitophagy,bone metabolism,osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells"in English and Chinese.The search time was from 2008 to 2023.According to the inclusion criteria,90 articles were finally included for review and analysis. RESULTS AND CONCLUSION:Mitophagy promotes the generation of osteoblasts through SIRT1,PINK1/Parkin,FOXO3 and PI3K signaling pathways,while inhibiting osteoclast function through PINK1/Parkin and SIRT1 signaling pathways.Mitophagy leads to bone loss by increasing calcium phosphate particles and tissue protein kinase K in bone tissue.Mitophagy improves the function of chondrocytes through PINK1/Parkin,PI3K/AKT/mTOR and AMPK signaling pathways.Modulation of mitophagy shows great potential in the treatment of bone diseases,but there are still some issues to be further explored,such as different stages of drug-activated mitophagy,and the regulatory mechanisms of different signaling pathways.

Aim To investigate how chili-derived exosome-like nanovesicle(CDELN)inhibited ox-LDL uptake and reduced ox-LDL-induced intracellular cholesterol accumulation.Methods CDELN were isolated and purified using tissue crushing,differential centrifugation,ultracentrifugation and sucrose density gradient centrifugation.Ox-LDL was used to stimulate THP-1-derived macrophages for 24 hours to establish a foam cell model in vitro,and the effect of CDELN on macrophage foam and its mechanism were further studied.Confocal laser microscopy was used to detect the uptake of CDELN and DiL-acetylated low density lipoprotein(DiL-ac-LDL)by THP-1 macrophages.Oil red O staining was used to detect intracellular cholesterol content and the positive area of oil red 0 staining in cells was analyzed to evalu-ate the effect of intracellular lipid accumulation.RT-qPCR and Western blot were used to detect mRNA and protein levels of scavenger receptor A(SRA)and cluster of differentiation 36(CD36),lectin-like oxidized low density lipoprotein recep-tor-1(LOX-1),ATP-binding cassette transporter A1/G1(ABCA1/G1).The expression of mitogen-activated protein ki-nase(MAPK)pathway proteins including p-ERK,p-p38 MAPK and p-c-Jun,were also analyzed.Results CDELN were exosome-like nanovesicles with uniform size and double membrane,rich in protein and nucleic acids,which can be taken up by macrophages.The results of DiL-ac-LDL uptake showed that CDELN could inhibit cholesterol uptake of mac-rophages.Oil red 0 staining showed that CDELN could reduce ox-LDL-induced intracellular cholesterol accumulation.RT-qPCR and Western blot showed that CDELN could significantly reduce mRNA levels of matrix metalloprotein-9(MMP-9),SRA,CD36 and LOX-1 and protein levels of p-ERK,SRA,CD36 and LOX-1 in ox-LDL-induced THP-1-derived macro-phages.Treatment with the p-ERK agonist Yoda1 diminished the protective effect of CDELN.Conclusion CDELN can significantly inhibit macrophage foam cell formation,and this effect may be associated with the inhibition of phosphoryl-ation levels of ERK1/2 in the MAPK pathway.

Depression is a mental disorder with high morbidity, disability and relapse rates. Ginkgo biloba extract (GBE), a traditional Chinese medicine, has a long history of clinical application in the treatment of cerebral and mental disorders, but the key mechanism remains incompletely understood. Here we showed that GEB exerted anti-depressant effect in mice through regulating gut microbial metabolism. GBE protected against unpredictable mild stress (UMS)-induced despair, anxiety-like and social avoidance behavior in mice without sufficient brain distribution. Fecal microbiome transplantation transmitted, while antibiotic cocktail abrogated the protective effect of GBE. Spatiotemporal bacterial profiling and metabolomics assay revealed a potential involvement of Parasutterella excrementihominis and the bile acid metabolite ursodeoxycholic acid (UDCA) in the effect of GBE. UDCA administration induced depression-like behavior in mice. Together, these findings suggest that GBE acts on gut microbiome-modulated bile acid metabolism to alleviate stress-induced depression.
Humans ; Mice ; Animals ; Depression/drug therapy* ; Gastrointestinal Microbiome ; Plant Extracts ; Ginkgo biloba

Objective To investigate the effect of p38MAPK gene silencing recombinant adeno-virus on the expression of target gene in different time and to detect the effect of p 38MAPK signal pathway on the upper lip scar hyperplasia at different time to determine the optimal scar treatment time .Methods The adenovirus vector was injected into the scar tissue in 0 week ,1 week and 2 week after cheiloplasty in rabbit .The specimens were harvested in 3 week postoperatively .Four methods in-cluding Sirius red staining ,immunohistochemical staining (IHC) ,Western blotting (WB) ,real-time PC (RT-PCR) were used to quantitatively and quantitatively detect the relative expression levels of p38MAPK and scar-related factors (col Ⅰ ,col Ⅲ ,MM P1 ,TIMP1) .Results Sirius red staining and immunohistochemical staining showed that in 1st week the expression of col Ⅲ and MMP1 in scar tis-sue was significantly higher than that in 0 week and 2 week after operation and the expression of col Ⅰand TIMP1 was significantly less than that in 0 week and 2 week after operation .The results of WB and RT-PCR were consistent with that of IHC .Conclusions After injection into the upper lip scar tis-sue with adenovirus in 1 week ,the degree of scar hyperplasia is the least .

Objective: To report on clinical characteristics and genetic findings in 15 Chinese patients with methylmalonic acidemia (MMA). Methods: For the 15 MMA patients detected by tandem mass spectrometry, genetic analysis was carried out in twelve pedigrees. Clinical characteristics, genetic finding, treatment and outcomes were retrospectively analyzed. Results: The main features of the patients included poor feeding, recurrent vomiting, lethargy, seizure and development retardation. Blood propionylcarnitine (except for 3 patients), its ratio with acetylcarnitine, and urine methylmalonic acid were increased in all patients. Twelve patients were diagnosed genetically, which included 7 with MUT variants, 4 with MMACHC variants, and 1 with MMAB variant. Nine MUT variants were detected, among which c. 1159A>C, 753+ 1delGinsTGGTTATTA and c. 504del were novel. Six known pathogenic MMACHC variants and two novel MMAB variants (c.289_290delGG, c. 566G>A) were also detected. Seven patients died of metabolic crises within a year, others had improved effectively following the treatment, but had mild to severe growth delay and/or developmental retardation. Conclusion The clinical manifestation of MMA are complex. Most patients have variants of the MUT and MMACHC genes. High mortality may occur before one year of age.

OBJECTIVERNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.

METHODSThe pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.

RESULTSSEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).

CONCLUSIONResults indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.

Animals ; Cleft Palate ; Gene Silencing ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; Oxidoreductases Acting on CH-CH Group Donors ; Palate ; growth & development ; RNA, Messenger

This article is aimed to provide a method for the simultaneous analysis of five heavy metals, including Cu, As, Pb, Cd, and Hg, in Chuanxiong Rhizoma and Gastrodiae Rhizoma through ICP-MS. Five heavy metals were determined by an inductively coupled plasma-mass spectrometry method after microwave-assisted digestion. The linear correlation coefficients were all better than 0.999. The lowest limits of quantification for all target elements were 0.003 0.134 ug·L-1, while the recovery values ranged from 80.04% 118.34%. This method was accurate, convenient, rapid, and highly sensitive, and can be applied to determine five heavy metals, including Cu, As, Pb, Cd, and Hg in Chuanxiong Rhizoma and Gastrodiae Rhizoma.

Objective To investigate the expression level of transforming growth factorβ1 (TGF-β1) and interleukin 17 (IL-17) in patients with allergic conjunctivitis. Methods TGF-β1 and IL-17 expression levels in allergic conjunctivitis patients hospitalized in the author's hospital and a healthy population were detected to explore the expression level in all types of patients with allergic conjunctivitis. Results TGF-β1 and IL-17 levels were significantly higher in patients with allergic conjunctivitis than in healthy population (P<0.01). Among the four clinical subtypes of allergic conjunctivi-tis, vernal keratoconjunctivitis had the highest expression level of TGF-β1 and IL-17 in spring, followed by perennial allergic conjunctivitis and seasonal allergic conjunctivitis, and atopic keratoconjunctivitis had the lowest expression level. However, the differences among the four subtypes were not significant (P>0.05). Conclusion Patients with allergic conjunctivitis have higher TGF-β1 and IL-17 expression levels than normal healthy people. TGF-β1 and IL-17 ex-pression level vary in different subtypes of allergic conjunctivitis, vernal keratoconjunctivitis has higher expression level than other types in spring.

Objective RNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves. Methods The pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV-siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n=10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n=10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experi-mental group (n=10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses. Results SEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves.RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a sig-nificant difference (P<0.05). Conclusion Results indicate that Dhcr7 gene silencing affects the fusion of palatal shelves.
Thus, Dhcr7 gene may serve a function in the normal development of palates.

The in-vitro fertilization and embryo transfer technique has been widely applied in human insemination. The rate of successful insemination is gradual y rising, and the in-vitro fertilization directly determine the insemination outcome.
OBJECTIVE:To evaluate the difference between the two common using insemination methods, microdrop and open, in in-vitro fertilization and embryo development.
METHODS:A randomized study was conducted to compare microdrop and open insemination methods among non-male factor patients undergoing in-vitro fertilization and embryo transfer. A total of 1 175 cases were enrol ed in the research. There were 573 cases in the microdrop group, and 602 cases in open insemination group. The fertilization rate and embryo development in the two groups were compared.
RESULTS AND CONCLUSION:The fertilization failure rate [total fertilization failure rate+low fertilization rate (<25%oocytes fertilized)] in the microdrop insemination group was higher than in the open insemination group (11.9%, 3.3%, P<0.001), while the good quality embryo rate and pregnancy rate did not differ significantly between the two groups (al P>0.05). The open insemination method is a simple insemination method with a lower fertilization failure rate. As the fertilization is a highly complicated process involving many extrinsic and intrinsic factors, further study is needed to confirm the effects of the two insemination methods on in-vitro fertilization outcome.