The expression of ferroptosis-related genes in osteoporosis based on GEO.

OBJECTIVE: The aim of this study was to screen the differential genes related to ferroptosis in osteoporosis patients. MATERIALS AND METHODS: GEO2R was used to screen the differential genes related to ferroptosis in osteoporosis patients by searching the relevant chips in the GEO database, and Spearman's correlation analysis was used to describe the correlation between quantitative variables without normal distribution. p-values lower than 0.05 were considered statistically significant. Another group of osteoporosis patients was selected in the GEO database to verify the significantly differentially expressed genes. RESULTS: The results showed that 10 samples in chip GSE35956 were identified as research objects, and a total of 5 ferroptosis differential genes were screened out: ATP5MC3, CDKN1A, MT1G, NCOA4, SLC1A5, of which 3 up-regulated genes (CDKN1A, MT1G, SLC1A5), 2 down-regulated genes (ATP5MC3, NCOA4). The above differential genes were placed in 19 samples of chip GSE35959 for verification, and the same expression trend was obtained, but only the MT1G difference was statistically significant. CONCLUSIONS: The gene correlation test found that MT1G and ATP5MC3 had a strong negative correlation.

Screening and bioinformatics analysis of senile osteoporosis genes based on GEO database.

OBJECTIVE: Despite improvements in research on osteoporosis in the elderly, the specific mechanism remains unknown. In order to develop better treatment regimens with better efficacy and fewer adverse reactions (ARs), it is vital to unravel the pathogenesis of osteoporosis in the elderly. The GEO chip was used to screen differential genes in senile osteoporosis and analyze their interaction mechanisms in order to obtain potential therapeutic pathways and targets. MATERIALS AND METHODS: GSE35956 was downloaded from GEO database and used as the research object for KEGG pathway enrichment analysis, GO enrichment analysis and protein-protein interaction (PPI) network analysis, respectively, to explore the related mechanisms of the occurrence and development of osteoporosis in the elderly. RESULTS: There were 156 differentially expressed genes in the elderly (72 years old) and middle-aged (42 years old) diagnosed with osteoporosis, of which 6 were up-regulated and 150 were down-regulated. An analysis of gene enrichment using GO (gene body) revealed that differentially expressed genes (DEG) were mainly distributed in extracellular matrix (ECM) and other cell structures. Its functions include ossification, parathyroid hormone metabolism, multicellular biological signaling pathway, vitamin catabolism, interleukisn-5 metabolism, transmembrane transporter activity, receptor signaling pathway, calcium metabolism and other molecular functions. According to the Kyoto Encyclopedia of Genes and Genomes (KEGG), an online resource, signaling pathways associated with age-related osteoporosis (OP) are significantly enriched. The DEG enrichment pathways include Wnt, ECM-receptor interaction, cGMP-PKG, GAG degradation, and calcium signaling. A protein and protein interaction (PPI) network was constructed for 14 key genes, including CD44, GRIA1, KNG1 and IL7R. CONCLUSIONS: The findings of this study indicate that CD44, GRIA1, KNG1, IL7R, and other differential genes affect the Wnt signaling pathway in the elderly, which can provide new targets for the follow-up basic research and treatment of osteoporosis in the elderly.

Clinical Feasibility of Using Single-isocentre Non-coplanar Volumetric Modulated Arc Therapy Combined with Non-coplanar Cone Beam Computed Tomography in Hypofractionated Stereotactic Radiotherapy for Five or Fewer Multiple Intracranial Metastases.

AIMS: To evaluate the clinical feasibility of single-isocentre non-coplanar volumetric modulated arc therapy (NC-VMAT) with non-coplanar cone beam computed tomography (NC-CBCT) in hypofractionated stereotactic radiotherapy (HSRT) for five or fewer multiple brain metastases. MATERIALS AND METHODS: Ten patients with multiple brain metastases who underwent single-isocentre NC-VMAT HSRT with limited couch rotations (within ±45°) and NC-CBCT with a limited scanning range (150-200°) were included in the current analysis. Conventional single-isocentre coplanar VMAT (C-VMAT) plans were generated and compared with NC-VMAT plans. The intracranial response and toxicities of single-isocentre NC-VMAT HSRT were also evaluated. RESULTS: Compared with C-VMAT, NC-VMAT generated better target conformity (P < 0.05), a lower gradient index (P < 0.05) and better normal brain tissue sparing, especially for volume ≥12 Gy, with a median reduction of 12.65 cm3. For 45° couch rotation, NC-CBCT produced sufficient image quality to differentiate bony anatomy, even with a 150° scanning range, which could be successfully used for patient set-up correction. After NC-CBCT, 57.1% of the measured non-coplanar set-up errors exceeded the threshold value. The median gamma passing rate of NC-VMAT was higher than that of C-VMAT plans (P < 0.05). The non-coplanar beam of NC-VMAT with NC-CBCT corrections exhibited superior gamma passing rate to that without NC-CBCT corrections. The intracranial objective response rate and disease control rate for all patients were 80% (8/10) and 100% (10/10), respectively, and the most common toxicities were headache (20%) and dizziness (20%). CONCLUSION: NC-VMAT with limited couch rotation (within ±45°) combined with NC-CBCT with a limited scanning range (150-200°) markedly improves the plan quality and set-up accuracy in single-isocentre multiple-target HSRT.

The effect of BMP2/Smads pathway mediating platelet-rich fibrin on rat bone mesenchymal stem cells.

OBJECTIVE: We explored the influences on platelet-rich fibrin (PRF) to rat Bone Mesenchymal Stem Cells (BMSCs), as well as the role of bone morphogenetic protein 2 (BMP2)/maternal signal protein homolog (Smads) pathway. MATERIALS AND METHODS: The proposed research is approved by the ethics board of the Second Affiliated Hospital of Harbin Medical University. The BMSCs were isolated and purified. The BMSCs were assigned to a control group arbitrarily, PRF group, BMP activator group and BMP inhibitor group (hereinafter referred to as activator group and inhibitor group). Each group of BMSCs in the logarithmic growth phase was detected for the alkaline phosphatase (ALP) activity since 3 days and 14 days of culture; CCK-8 assay was conducted for detection of the proliferation of BMSCs; Real time PCR was conducted for detection of the osteogenic differentiation marker collagen I (COL-I), BMP2, Runt-related transcription factor 2(RUNX2), osteocalcin (OCN) mRNA relative expression levels; Western-Blot detection of BMP2, OCN, P-SMAD1/5/8, relative expression level of RUNX2 protein. RESULTS: In contrast to the control group, BMSCs' the ALP activity of the PRF group, activator group, as well as inhibitor group increased for 3 days and 14 days, and the activator group>PRF group>inhibitor group (p≤0.05). ALP activity in each group was elevated with the increase in culture time, the ALP activity of the control group, PRF group, activator group and inhibitor group increased (p≤0.05). In comparison to the control group, the relevant expression levels of COL-I, BMP-2, RUNX2 and OCN in the PRF group, activator group, and inhibitor group increased, and the activator group>PRF group>inhibitor group (p≤0.05). The relative expression levels of BMP2, OCN, p-SMAD1/5/8 and RUNX2 protein in each group were statistically different, the activator group>PRF group>control group>inhibitor group (p≤0.05). CONCLUSIONS: PRF can promote the proliferation and osteogenic differentiation of BMSCs by activating the BMP2/Smads signaling pathway.

[Mechanism of paeoniflorin inhibiting apoptosis of hippocampal neurons of rats induced by lead acetate].

Objective: To investigate the effect and underlying mechanism of paeoniflorin on hippocampal neuron apoptosis induced by lead acetate. Methods: In September 2020, primary hippocampal neuronal cells were isolated and cultured from fetal rats, and identified using cellular immunofluorescent. MTT assay was used to measure the cell viability to determine the concentration and time of lead acetate-induced hippocampal neuron apoptosis. MTT was also used to evaluate the effect of paeoniflorin concentration on the apoptosis of hippocampal neurons induced by lead acetate. According to the results, different concentrations of paeoniflorin were selected to intervene hippocampal neuron cells, after 24 h, lead acetate was added to the cells, meanwhile, blank and model groups were set up, the content of reactive oxygen species (ROS) , superoxide dismutase (SOD) , lactate dehydrogenase (LDH) , malondialdehyde (MDA) and Caspase-3 were measured. Extracellular signal regulated kinase (ERK) , phosphorylated ERK (p-ERK) , p38 mitogen -activated protein kinases (p38MAPK) , phosphorylated p38MAPK (p-p38MAPK) , c-Jun N-terminal kinase (JNK) and phosphorylated JNK (p-JNK) protein expression in hippocampal neuronal cells were determined by Western blotting. Results: The isolated and cultured hippocampal neurons were identified by immunofluorescence chemical staining and then treated with lead acetate, MTT results showed that lead acetate had the best toxicity effect when treated for 24 h at a concentration of 25 µmol/L. Paeoniflorin showed no cytotoxic effect on hippocampal neuronal cells when the concentrations below 80 µmol/L. Compared with the model group, the activity of hippocampal neuronal cells was significantly increased after treating with 20, 40 or 80 µmol/L paeoniflorin (P<0.05) . Compared with the blank group, the ROS activity, LDH release level, MDA content and caspase-3 content were significantly increased (P<0.01) , and the SOD activity was significantly decreased (P< 0.01) in the hippocampal neuronal cells of the model group. Compared with the model group, the ROS activity, LDH release level, MDA content and caspase-3 content were obviously decreased (P<0.05) , SOD activity was significantly increased (P <0.01) after hippocampal neuronal cells were treated with 40 or 80 µmol/L paeoniflorin. Relative to the model group, the ratio of p-ERK/ERK were significantly up-regulated (P<0.01) , while the ratios of p-p38MAPK/p38MAPK and p-JNK/JNK were significantly down-regulated after hippocampal neuronal cells were treated with 40 or 80 µmol/L paeoniflorin (P<0.05) . Conclusion: Paeoniflorin may down-regulate the expression of p-p38MAPK and p-JNK protein, up-regulate the expression of p-ERK protein, and inhibit the apoptosis of hippocampal neurons induced by lead acetate through the MAPK signaling pathway.

Long non-coding RNA ROR is a novel prognosis factor associated with non-small-cell lung cancer progression.

OBJECTIVE: The aim of the present study was to determine the expression levels of long intergenic non-protein coding RNA, regulator of reprogramming (linc-ROR) in non-small-cell lung cancer (NSCLC) patients and to further explore the prognostic value of this lncRNA. PATIENTS AND METHODS: In our investigation, we determined the expression of linc-ROR in human NSCLC tissues and matched normal lung tissues by quantitative Real-time-PCR analysis. Also, correlations between linc-ROR expression and the clinicopathological features were evaluated. Survival curves were plotted using the Kaplan-Meier method and differences in survival rates were analyzed using the log-rank test. Cox regression analyses were performed to explore the effect of linc-ROR as an independent predictor of survival. RESULTS: We found that linc-ROR had high expression in NSCLC specimens than that in matched adjacent normal lung tissues (p < 0.01). In addition, higher linc-ROR expression levels were positively correlated with advanced TNM stage (p = 0.007), positive distant metastasis (p = 0.001) and LN metastasis (p = 0.011). Furthermore, significantly shorter 5-year overall survival (OS) and disease-free survival (DFS) were observed in patients with higher expression of linc-ROR (both p < 0.001). In a multivariate Cox model, it was found that linc-ROR expression was an independent prognostic factor for both 5-years OS (p = 0.001) and 5-year DFS (p = 0.001) in NSCLC. CONCLUSIONS: Our findings indicate that linc-ROR plays an oncogenic role in NSCLC development and may function as a prognostic and predictive biomarker for NSCLC.

[Experimental study on the expression of α-SMA and HMGB1 in silicotic fibrosismodel rats interfered by lumbricus].

Objective: To observe the expression of alpha smooth muscle actin (α-SMA) and high mo-bility group protein B1 (HMGB1) in silicosis model rats interfered by lumbricus. Methods: 45 rats were ran-domly divided into the control group, model group and group interfered by lumbricus. The silicosis model rats were established. The group interfered by lumbricus were intragastric administered with lumbricus decoction by the 4 ml/kg dose. The control group and model group were ig administered with the equal amount of normal saline. Each group were killed 5 rats on the 7(th), 14(th) and 28(th) day. The lung tissues were stained with HE and Sirius red methods. The mRNA expressions of α-SMA and HMGB1 were determined with RT-PCR; The pro-tein levels of α-SMA and HMGB1 were determined with Western blotting. Results: Compared with the control group, the expression levels of α-SMA and HMGB1mRNA and protein in lung tissue of model group were grad-ually increased in the 7(th), 14(th) and 28(th) days, the difference was statistically significant (P< 0.01) . Compared with model group, the levels of α-SMA and HMGB1mRNA and protein in lung tissue of group interfered by lumbricus were gradually lowered in the 7th, 14th and 28th days, the difference was statistically significant (P<0.05, P<0.01) . Conclusion: Lumbricus inhibits the collagen deposition and the formation of silicosis pulmo-nary fibrosis, which may be related to the inhibition of HMGB1 expression and activation of α-SMA in lung tis-sue.

[The study of the eosinophil CD34+ progenitor cells differentiation mechanism of model rats with occupational asthma and the intervention of warm and tonifying kidney yang decoction].

Objective: To investigate the mechanism of CD34+ progenitor cell differentiation in rat by observing the change relations between the eosinophils (EOS) and the content of Eotaxin and IL-5 in blood and the CD34+/CCR3+, CD34+/IL-5Rα+ in bone marrow after occupational asthma (OA) model rats are simulated, and to observe the effect of WTKYD Trraitional Chinese Medicine intervention. Methods: A total of 40 healthy male SD model rats (200~250 g weight) were randomly divided into model contrast Group, prednisone acetate intervention Group, WTKYD+1/2 prednisone acetate intervention Group and WTKYD intervention Group, 10 in each group, and set a Group for blank contrast. Give them saline (20 ml/kg) , prednisone acetate (8.22 mg/kg) , WTKYD (20g/kg) +1/2 prednisone acetate (4.11 mg/kg) and WTKYD (20 g/kg) intervention respectively. By means of cell count, immunohistochemical, ELISA, flow cytometry technique, situ hybridization and so on, to observe EOS and the expression of Eotaxin in lung tissue, the EOS in peripheral blood, the content of Eotaxin and IL-5 in blood as well as the expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in bone marrow respectively. Results: The number of EOS, the content of Eotaxin and IL-5, the expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in Model Contrast Group were higher in Blank Contrast Group, the difference was statistically significant (P<0.01) , while they were lower in medical intervention Groups when comparing to Model Contrast Group, the difference was statistically significant (P<0.01 or P<0.05) , and the above items in WTKYD+1/2 Prednisone Acetate Intervention Group were even lower than in Prednisone Acetate Intervention Group and WTKYD Intervention Group, the difference was statistically significant (P<0.05). EOS in lung tissue is highly positive related to the content of Eotaxin and IL-5 in peripheral blood as well as the expression of CD34+/CCR3 and CD34+/IL-5Rα in bone marrow (0.9666、0.9829、0.9142, 0.8874). Conclusion: The increase of internal EOS in lung tissue is related to the up-regulated expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in bone marrow after antigens in Occupational Asthma model rats are stimulated. Through down-regulating it's expression to restrain the differentiation of CD34+ progenitor cells towards EOS, meanwhile, the collaboration of WTKYD and prednisone acetate possess a certain synergistic action.

[Association of single nucleotide polymorphisms in PAI-1 with breast cancer susceptibility and prognosis].

OBJECTIVE: To study the effects of single nucleotide polymorphisms (SNP) in Plasminogen activator inhibitor 1(PAI-1) on breast cancer susceptibility and patients' prognosis among a Chinese Han women population. METHODS: Six tag SNP (tSNP) of PAI-1 were selected according to HapMap CHB population, and TaqMan realtime PCR method was used to genotype the 6 tSNP in 1 160 breast cancer cases and 1 318 age-matched controls among Chinese Han women. Haplotypes and diplotypes were inferred according to genotyping data and linkage disequilibrium. Finally, the associations of tSNP, haplotypes and dipltypes with breast cancer susceptibility and patients' prognosis were analyzed. RESULTS: Regarding to breast cancer susceptibility, for rs6090 (G>A), AA genotype carriers had 3.79 times higher risk of developing breast cancer (OR=4.79, 95%CI=1.01-22.64, P=0.048 0) than GG or GA genotype carriers. For rs2227672 (G>T), TT genotype carriers had 1.52 times higher breast cancer risk than GG or GT genotype carriers (OR=2.52, 95%CI=1.26-5.01, P=0.008 6). Regarding to breast cancer prognosis, women who carried rs2227692 (C>T) CT genotype had 46% lower risk of developing recurrence, metastasis or death than CC genotype carriers (HR=0.54, 95%CI=0.30-0.97, P=0.040 4). Using stratified association analysis, among BMI<23 patients, those women who carried AA genotype of rs2227631 (G>A) had 3.99 times higher risk of developing the events (recurrence, metastasis or death) than GG or GA genotype carriers (HR=4.99, 95%CI=1.66-15.02, P=0.004 2). Among HER2 positive patients, those women who carried AA genotype of rs2227667 (G>A) had 2.98 times higher risk of developing the events (recurrence, metastasis or death) than GG or GA genotype carriers (HR=3.98, 95%CI=1.47-10.80, P=0.006 7). Among patients with tumors>2 cm, those women who carried rs2227692 (C>T) CT or TT genotype had 51% lower risk of developing the events (recurrence, metastasis or death) than CC genotype carriers (HR=0.49, 95%CI=0.27-0.88, P=0.017 0). CONCLUSIONS: The study indicates that single nucleotide polymorphisms in PAI-1 may affect breast cancer susceptibility and survival in Chinese Han women. The study may contribute to individualized evaluation of breast cancer risk and patients' prognosis if these data are validated in some other Chinese Han populations.

[215 cases of diffuse pulmonary disease diagnosed with transbronchial lung biopsy via bronchofibroscope].

215 cases of diffuse pulmonary disease diagnosed with transbronchial lung biopsy (TBLB) via bronchofibroscope were reported. The incidence of successful biopsy was 96.7%. The total diagnostic rate was 80.5%. There were no other serious complications except for bloody or blood-streaking sputum, but the amount rarely exceeded 50 ml, and pneumothorax occurred occasionally. It was suggested that TBLB is an easy, safe and useful method for diagnosing diffuse pulmonary disease.

[The diagnosis and treatment in 39 cases of sarcoidosis].

The diagnosis, treatment and follow-up in 39 cases of sarcoidosis were reported. The authors believed that (1) diagnosis of this disease would be made by analysing clinical and pathological manifestations carefully, (2) treatment was given when the disease was active, and (3) in order to find complications as early as possible, follow-up was very important during treatment.

Pneumoconiosis: the changes of peripheral airways.

Ultrathin bronchofiberscope (Olympus BF-1.8T) was applied to the observation of the peripheral airways of total 87 cases, including 73 cases of quartz-exposed and asbestos-exposed workers with/without pneumoconiosis (exposure groups), and 14 cases of controls. The changes of peripheral airways in exposure group of non-smokers was found, mainly to be pigmentation, elevation, irregularity and lustlessness on the surface of the bronchiole walls, and predominantly stenosis and ectasis of the lumen. The detectable rate was higher in peripheral airways than in central airways. Some changes existed in early stage and some had a tendency of increasing detectable rate with category progressing. Some factors such as smoking could influence the results of analysis.

Ten families of ankylosing spondylitis.

Twenty-two patients with ankylosing spondylitis (AS) from 10 families were studied with special attention to their clinical findings, HLA-B27 examinations and family histories. Results showed that HLA-B27 was positive in 19 and negative in 3. The hereditary relationship between ankylosing spondylitis and HLA-B27 in Chinese was similar to that in westerners. We consider that subjective symptoms and clinical findings are essential to early diagnosis of AS, but HLA-B27 examination and family history are supportive.