Objective Explore the relationship between semen quality and trace element levels in seminal plasma,as well as miR-184 levels in exosomes of male infertility patients.Methods 155 male infertility patients in our hospital from January 2022 to July 2023 were selected as the infertility group,and another 155 male patients with normal physical examination during the same period were selected as the normal group.Compare the general information,semen quality parameters,trace elements,and miR-184 levels in exosomes between two groups,and analyze the correlation between male infertility patients and the above laboratory indicators through Spearman rank correlation analysis.Results The sperm concentration,total sperm motility(PR + NP),forward motility sperm(PR),average curve velocity,average linear velocity,average path velocity,and average lateral amplitude of sperm head(ALH)in the infertile group were lower than those in the normal group(P<0.05);The levels of Zn,Mg,and Ca in the infertile group were lower than those in the normal group,while the levels of Cu,Pb,and relative expression of miR-184 were higher than those in the normal group(P<0.05);The levels of Zn and Cu elements in male infertility patients are positively and negatively correlated with sperm concentration,total sperm motility(PR+NP),and forward motility sperm(PR),respectively.The levels of Pb elements are negatively correlated with the average lateral amplitude of sperm head(ALH),and P<0.05.The relative expression level of miR-184 in male infertility patients is negatively correlated with sperm concentration,total sperm motility(PR + NP),forward motility sperm(PR),and average sperm curve velocity,with P<0.05.Conclusion The quality of semen in male infertility patients is closely related to the levels of trace elements in semen and miR-184 levels in exosomes.

BACKGROUND:Exosomes are vesicle-like structures secreted by cells into extracellular compartments in the form of cytosol,which contain a large amount of microRNAs with important intercellular communication roles.MicroRNAs in exosomes rely on exosome transport and are able to enter target cells to exert important biological regulatory effects.In common bone and joint diseases,abnormal or damaged bone metabolism releases a large number of exosomes,while some exosome-derived microRNAs also promote the progression of osteoarthritis.Therefore,exosome-derived microRNAs are closely related to the skeletal system and are important for the development as well as diagnosis and treatment of many osteoarticular diseases. OBJECTIVE:To review the research progress of exosome-derived microRNAs in bone metabolism and bone and joint diseases. METHODS:Using"exosomes,extracellular vesicle,microRNA,miRNA,bone,bone diseases,bone formation,bone regeneration,bone resorption,bone destruction"as Chinese and English search terms,articles were searched on CNKI,Metasys,and PubMed databases.Finally,86 articles were included for summarization. RESULTS AND CONCLUSION:Exosome-derived microRNAs can regulate bone metabolism by affecting bone formation and bone resorption,and are closely related to the development of bone and joint diseases such as fracture healing,osteoporosis,osteoarthritis,rheumatoid arthritis,osteonecrosis of the femoral head,and osteosarcoma.Exosome-derived microRNAs will be an effective means of diagnosis and treatment of certain bone and joint diseases in the future.However,the current research on exosome-derived microRNAs in osteoarthritic diseases is limited,and more explorations and researches are still needed to diagnose and treat osteoarthritic diseases using exosome-derived microRNAs.

OBJECTIVE: To explore the genetic basis for a couple with normal phenotype but repeated pregnancies with fetuses affected by osteogenesis imperfecta. METHODS: Whole exome sequencing (WES) was carried out on fetal specimens and parental DNA to detect potential pathologic variants. Suspected variants were verified by Sanger sequencing. Semen sample of the husband was collected for the extraction of genome DNA, and whole genome amplification (WGA) was performed for single sperms isolated from the sample. RESULTS: WES has identified a heterozygous c.1378G>A (p.G460S) variant of the COL1A2 gene in the fetus, which was predicted to be pathogenic but not detected in peripheral blood samples of both husband and wife. The heterozygotic variant was detected in semen DNA from the husband. Among 15 spermatozoa, 4 were found to harbor the variant. CONCLUSION The fetus was diagnosed with osteogenesis imperfecta, and the gonadal mosaicism probably accounted for the repeated abnormal pregnancies. Possibility of gonadal mosaicism should be considered when counseling couples with normal phenotype and genotype but recurrent abnormal pregnancies and/or births of children with similar phenotypes and genetic variants.
Adult ; Child ; Collagen Type I ; genetics ; Female ; Fetus ; Gonadal Disorders ; genetics ; Humans ; Male ; Mosaicism ; Mutation ; Osteogenesis Imperfecta ; diagnosis ; genetics ; Pregnancy ; Prenatal Diagnosis ; Whole Exome Sequencing

Objective: To explore the inhibitory effect of exosomes secreted by human umbilical cord mesenchymal stem cells(HUCMSC) on apoptosis of human umbilical vein endothelial cells(HUVEC) after model group(oxygen-glucose deprivation reoxygenation), and to clarify its possible mechanism. Methods: Human umbilical cord mesenchymal stem cells were cultured. The collected cell supernatant was stored in a centrifugal tube. The exosomes secreted by human umbilical cord mesenchymal stem cells were extracted by ultracentrifugation and identified. Human umbilical vein endothelial cells were randomly divided into control group, model group and different concentrations of HUCMSC-EXO(20 μg/ml, 40 μg/ml, 60 μg/ml) treatment groups(adding HUCMSC-EXO into the model group) . The morphological changes of HUVEC cells in each group were observed by inverted phase contrast microscope, and the proliferation inhibition rate of HUVEC in each group was measured by CCK-8 reagent. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3, Bax, Bcl-2 and hypoxia-associated protein hypoxia inducible factor 1α(HIF-1α). Inhibitor(HIF-1α inhibitor) + model group and HUCMSC-EXO + inhibitor + model group were added on the basis of the above experiments. Western blot analysis was performed to observe the effects of HUCMSC-EXO, inhibitor and both of them on HIF-1α and Bax expressions in HUVEC. Results: HUCMSC-EXO was successfully extracted and identified. Compared with the control group, the volume of HUVEC in the model group and the HUCMSC-EXO group with different concentrations decreased, became round, connected and evacuated, and the growth state was poor under the inverted phase contrast microscope.CCK-8 detection showed that the cell viability in the HUCMSC-EXO group was significantly higher than that in the model group, the difference was statistically significant (t=9.23, P<0.05). Western blot analysis showed that compared with the control group, the expression levels of Caspase-3 ((0.296±0.038), (0.879±0.088); t=14.92, P<0.05), Bax((0.234±0.034), (0.762±0.084); t=14.36, P<0.05) of HUVEC in the model group were up-regulated, and the expression level of Bcl-2 was down-regulated ((0.863±0.103), (0.387±0.059); t=9.85, P<0.05), with statistically significant differences. Compared with the model group, the expression levels of Caspase-3( (0.586±0.075); t=6.24, P<0.05), Bax((0.311±0.055); t=11.01, P<0.05) and Bcl-2((0.665±0.071); t=7.45, P<0.05) of HUVEC in the HUCMSC-EXO treatment group were down-regulated and the differences were statistically significant. Inhibitor intervention experiments showed that there were no significant differences between the inhibitor+ model group and HUCMSC-EXO+ inhibitor+ model group in the expression of HIF-1α protein ((0.348±0.055), (0.388±0.077); t=1.04, P>0.05)and Bax protein ((0.363±0.069), (0.370±0.064); t=0.18, P>0.05). But both of them were down-regulated compared with the model group (HIF-1α protein (0.919±0.064), Bax protein (0.902±0.071)), the differences were significant( t=13.56, t=13.03, both P<0.05). Conclusion HUCMSC-EXO has a protective effect on OGD/R model of HUVEC, and its mechanism may be related to the down-regulation of HIF-1α expression.

Intertrochanteric fracture of femur (IFF) is a common fracture type in traumatic orthopedics,which seriously affects patients' quality of life and health.Studies have shown that the fibrinolytic system can be activated in the invasive operation of IFF,which leads to massive blood loss during and after operation,especially hidden blood loss.Tranexamic acid,as an anti-fibrinolytic drug,can effectively reduce fibrinolytic activity by inhibiting fibrinolysis induced by fibrinolytic enzymes in order to stop bleeding.In recent years,with the diversified development of surgical hemostasis methods and the continuous exploration of hemostatic drugs,the application of anti-fibrinolytic drugs represented by tranexamic acid in perioperative period of IFF has gradually increased.Based on literatures at home and abroad,this paper aims to summarize the research progress of the application of tranexamic acid to reduce the perioperative hidden blood loss in intertrochanteric fracture,providing reference for clinical treatment decision.

Objective To observe the expression of neuropilin-1 (NRP-1) in glioma cells of different grades,and evaluate the application value of a novel molecular probe(USPIO-PEG-tLyP-1)in the grading diagnosis of heterotopic glioma in nude mice by magnetic resonance imaging (MRI).Methods Expression levels of NRP-1 in glioma cell lines of different grades were detected by Western-Blot.USPIO-PEG-tLyP-1 was synthesized by carbon diimine method.The U87-MG tumor-bearing mice model (U87-MG group) and CHG-5 tumor-bearing mice model(CHG-5 group) were established with 10 mice in each group.Six tumorbearing mice with a tumor volume about 0.6 cm3 were selected from each group,and they were given with 2mg/kg molecular probes via tail vein respectively and was detected by MRI at 0 h,6 h,12 h and 24 h,then R2 values were calculated.After the imaging,tumor-bearing mice were sacrificed,and tumor tissue sections were made.The iron particles in the sections was detected by Prussian blue staining.The binding ability of molecular probes and tumor tissues in the two groups was compared.Results The expression of NRP-1 in U87-MG and CHG-5 cell lines was significantly higher than that in HA.In addition,the expression of NRP-1 in U87-MG was higher than that in CHG-5 cell(P＜0.01).MRI results showed that R2 values of tumor tissues in the two groups were compared,and the difference was not statistically significant before the injection of molecular probe(U87-MG group(10.35±0.52)vs CHG-5 group(9.86±0.43),t=1.779,P=0.106).The R2 value of tumor tissue in the U87-MG group was higher than that in the CHG-5 group after the injection of molecular probe (6 h:U87-MG group (11.63±0.85)vs CHG-5 group (10.51 ±0.49),t=2.796,P=0.019;12h:U87-MG group(14.23±0.68)vs CHG-5 group(12.29±0.28),t=6.462,P=0.000;24 h:U87-MG group (13.36±0.92) vs CHG-5 group(11.32±0.64),t=4.459,P=0.001).The results of Prussian blue staining showed that there were significantly more blue staining particles in tumor tissues of the U87-MG group than that of the CHG-5 group,and the difference was statistically significant(P＜0.01).Conclusion The NRP-1 targeted molecular probe can be used for grading diagnosis of high and low grade heterotopic brain glioma in nude mice.

Objective To explore the clinical,imaging,genetic features in a case of fatal familial insomnia (FFI),and review related literatures.Methods A case of middle-aged woman diagnosed as frontotemporal dementia based on the preliminary manifestation of abnormal mental behavior was reported.The clinical features,imaging characteristics,electroencephalogram and polysomnogram of the patient were analyzed,and the blood samples from the patient and some of her familial members were collected for the sequencing of prion protein gene (PRNP).Results This patient was a middle-aged woman,whose clinical manifestations were abnormal mental behavior,rapid progressive dementia and intractable insomnia,abnormal night sleep behavior and laryngeal stridor.Brain MRI indicated frontotemporal lobe atrophy.Non-sleep disturbance was observed in polysomnography.The cerebrospinal fluid was negative for 14-3-3 protein.The results of PRNP sequencing revealed that the mutation of gene D178N/129M was detected.Conclusions Detection of PRNP plays an important role in the diagnosis of FFI.Patients suspected of FFI in clinic should be detected for genetic testing.Whether the frontotemporal lobe atrophy was caused by FFI or concurrent with FFI remains to be further verified.

Werner syndrome (WS) is a premature aging disorder that mainly affects tissues derived from mesoderm. We have recently developed a novel human WS model using WRN-deficient human mesenchymal stem cells (MSCs). This model recapitulates many phenotypic features of WS. Based on a screen of a number of chemicals, here we found that Vitamin C exerts most efficient rescue for many features in premature aging as shown in WRN-deficient MSCs, including cell growth arrest, increased reactive oxygen species levels, telomere attrition, excessive secretion of inflammatory factors, as well as disorganization of nuclear lamina and heterochromatin. Moreover, Vitamin C restores in vivo viability of MSCs in a mouse model. RNA sequencing analysis indicates that Vitamin C alters the expression of a series of genes involved in chromatin condensation, cell cycle regulation, DNA replication, and DNA damage repair pathways in WRN-deficient MSCs. Our results identify Vitamin C as a rejuvenating factor for WS MSCs, which holds the potential of being applied as a novel type of treatment of WS.
Animals ; Ascorbic Acid ; pharmacology ; Cell Cycle Checkpoints ; drug effects ; Cell Line ; Cellular Senescence ; drug effects ; DNA Damage ; DNA Repair ; drug effects ; DNA Replication ; drug effects ; Disease Models, Animal ; Heterochromatin ; metabolism ; pathology ; Humans ; Mesenchymal Stem Cells ; metabolism ; pathology ; Mice ; Nuclear Lamina ; metabolism ; pathology ; Reactive Oxygen Species ; metabolism ; Telomere Homeostasis ; drug effects ; Werner Syndrome ; drug therapy ; genetics ; metabolism

Objective To study the value of serum tartrate-resistant acid phosphatase 5b (TRACP5b) in bone metastasis in early breast cancer patients with negative axillary lymph nodes.Methods The serum level of TRACP5b was determined by enzyme-linked immumosorbent assay (ELISA) in 26 breast cancer patients and 18 healthy female adults.Results The serum level of TRACP5b was (3.85 ± 0.85) U/L in the group of breast cancer patients and (2.08 ± 0.84) U/L in the control group.The difference had statistical significance (P ＜0.05 ).The sensitivity was 42.31％ and the specificity was 94.44％ for TRACP5 b in predicting bone metastasis of breast cancer with negtive axillary lymph nodes.Conclusions TRACP5b has higher sensitivity and specificity in breast cancer patients than in healthy female adults.TRACP5b is a useful serum marker to predict bone metastasis in breast cancer patients.

Purpose: To observe therapeutic effect of acupuncture plus Methadone on improvement of heroin withdrawal syndrome. Methods: Sixty cases of heroin dependence patients were divided into the treatment group and the control group. The treatment group was given acupuncture plus Methadone, and the control group was treated with Methadone.Results: The score of the withdrawal symptoms was lower in the treatment group than in the control group (P ＜ 0.05). The improvement of withdrawal symptoms was significantly better in the treatment group than in the control group (P＜ 0.05). Conclusion: Acupuncture plus Methadone can significantly improve heroin withdrawal syndrome.