ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism. MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2. ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05). ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.

Objective: To investigate the effects of "liver-smoothing and spirit-regulating" acupuncture on the intestinal flora, lipopolysaccharide (LPS) and the hippocampal toll-like receptor 4 (TLR4)/ transcription factor (NF)-κB signaling pathway in depressive disorder mouse model, and to explore its underlying mechanisms. Methods: Eighteen male SPF-grade C57BL/6J mice were randomly assigned to the control, model, and acupuncture groups using a random number method, with six mice in each group. The depression disorder model was induced in mice from both the acupuncture and model groups using CUMS. The mice in the acupuncture group were treated with acupuncture at the acupoints of "Baihui" (DU20), "Yintang" (DU29), "Hegu" (LI4), and "Taichong" (LR3) on the 15th day of modeling, with a duration of 20 min per session, once per day, for 2 consecutive weeks. Behavioral differences were assessed using the sucrose preference test, open field test, and forced swim test. Hematoxylin-eosin staining was used to observe pathological changes in the hippocampus and colon. The levels of the inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and LPS in the hippocampus and colon were measured using Enzyme-linked Immunosorbent Assay. Western blotting was used to detect the expression of TLR4 and NF-κB protein in the hippocampus. Changes in gut microbiota structure and abundance were analyzed by 16 S rDNA sequencing. Results: Compared to the control group, the model group showed reduced sucrose preference rate, time in the center area, and total distance, with an increase in immobility time (P<0.01). Inflammatory pathological changes were observed in the hippocampal CA1 region and colon. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon increased (P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were increased (P<0.01). The Chao1 index was increased (P<0.01). The relative abundances of Pseudomonadales, Acinetobacter, Moraxellaceae, Solibacillus, Escherichia_shigella, Enterobacteriaceae, Enterobacterales, Dubosiella, and Erysipelottichales were decreased, while the relative abundances of Alloprevotella and gram_negative_bacteriurh_cTPY_13 were increased (P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were upregulated, and the pathway of terpenoid backbone biosynthesis was downregulated (P<0.01). Compared to the model group, the acupuncture group showed increased sucrose preference, time in the center area, and total distance, with a decrease in immobility time (P<0.01). The inflammatory pathological changes in the hippocampal CA1 region and colon were alleviated. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon were reduced(P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were reduced (P<0.01). The Chao1 index was decreased (P<0.05), and the relative abundances of Dubosiella and Erysipelotrichaceae were increased, while the relative abundance of Rikenellaceae, Alloprevotella, and gram_negative_bacteriuch_cTPY_13 were decreased(P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were significantly downregulated, and the pathway of terpenoid backbone biosynthesis was upregulated (P<0.01). Conclusion " Liver-smoothing and spirit-regulating" acupuncture can improve depressive symptoms in depressive disorder mice, potentially through regulating the LPS and TLR4/NF-κB signaling pathway mediated by intestinal flora, reducing the inflammatory response of the hippocampus, and improving the pathological injury of the hippocampus.

Objective To investigate the effects of microRNA(miRNA,miR)-887-3p on the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells and its underlying molecular mechanism.Methods Annulus fibrosus tissues were obtained from 8-week-old SPF-grade SD male rats,centrifuged to prepare and identify annulus fibrosus cells.Rats in the experiment were randomly divided into four groups:a Normal group consisting of primary annulus fibrosus cells without any treatment;a Control group treated with 10 ng/mL interleukin-1β(IL-1β)for 24 hours to establish a degenerative cell model;an interference group(miR-887-3p inhibitor)transfected with miR-887-3p inhibitor using Lipo3000 based on the Control group;and an overexpression group(miR-887-3p mimics)transfected with miR-887-3p mimics using Lipo3000 based on the Control group.CCK-8 assay was used to assess cell viability;flow cytometry was used to measure cell apoptosis rates;real-time fluorescence quantitative PCR(qPCR)was used to detect the expression levels of miR-887-3p and murine double minute 4(MDM4)mRNA;Western blotting was used to measure the protein expression levels of MDM4,Bcl-2,and Caspase-3.Results Immunofluorescence staining of isolated and cultured cells revealed a Collagen I positive rate of over 90％in rat intervertebral disc annulus fibrosus cells,indicating a cell purity level greater than 90％.Real-time fluorescence qPCR results showed that after establishing an annulus fibrosus degenerative cell model using IL-1β,the expression level of miR-887-3p significantly increased compared to the Normal group(P＜0.001).Compared to the Control group,transfection with miR-887-3p inhibitor resulted in a significant decrease in its expression level(P＜0.001).The CCK-8 assay showed that compared to the Normal group,cell viability significantly decreased in the Control group(P＜0.001).Compared to the Control group,cell proliferation ability significantly increased after miR-887-3p inhibition,and significantly decreased after overexpression of miR-887-3p.Flow cytometry results revealed that compared to the Normal group,the apoptosis rate in the Control group significantly increased(P＜0.001).Compared to the Control group,the cell apoptosis rate significantly decreased in the miR-887-3p interference group(P＜0.001)and increased in the overexpression group(P＜0.001).Western blotting analysis showed that compared to the Normal group,Bcl-2 expression level significantly decreased(P＜0.001)and Caspase-3 expression level significantly increased(P＜0.001)in the Control group.Compared to the Control group,Bcl-2 and MDM4 expression levels significantly increased(P＜0.01),and Caspase-3 expression level significantly decreased(P＜0.01)in the miR-887-3p interference group;whereas in the overexpression group,Bcl-2 and MDM4 expression levels significantly decreased(P＜0.05),and Caspase-3 levels significantly increased(P＜0.05).Real-time fluorescence qPCR and protein immunoblotting results showed that after interfering with miR-887-3p,the expression of MDM4 protein and mRNA increased(P＜0.001);after overexpressing miR-887-3p,their expression decreased(protein,P＜0.01;mRNA,P＜0.001).Conclusion MiR-887-3p may modulate the cell proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells by regulating MDM4 expression,thereby influencing the development and progression of disc degeneration.

Objective:Biological markers of the ventrolateral orbitofrontal cortex(vlOFC)involved in pain regula-tion were screened.Methods:Chronic inflammatory pain was induced in male C57BL/6J mice by injection of complete Freund's adjuvant(CFA)into the left posterior plantar.Paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were detected to evalue hyperalgesia.Transcriptome sequencing was performed on fresh tissue from vlOFC of mice after behavioral tests.The differentially expressed genes(DEGs)were screened by bioinformatics method,and their biological functions and pathways were enriched.Results:Compared with the PBS group,the left hindpaw me-chanical pain threshold and the paw withdrawal latency caused by heat pain were significantly reduced in the CFA group(P＜0.001).The DEGs of vlOFC in the two groups were 497,of which 143 were up-regulated and 354 were down-reg-ulated.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGGs)analysis showed that:In chro-nic inflammatory pain model mice,DEGs of vlOFC were mainly manifested in biological processes such as organic cation transport,neurotransmitter transport,and regulation of cytoplasmic calcium ion concentration.It is related to G protein-coupled receptors(GPCRs),neuropeptides and ammonium transport.DEGs mainly focuses on neuroactive ligand-receptor interactions,cytokine-cytokine receptor interactions,and cAMP signaling pathways.Reactome functional en-richment analysis showed that the pathway with the highest number of DEGs enriched and the lowest P value-adjusted was GPCRs ligand binding.Conclusion:Ion transport,neurotransmitter transport and binding,and GPCRs-related ac-tivities in vlOFC are involved in the regulation of chronic inflammatory pain.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective:To systematically evaluate the risk factors for postoperative pulmonary complications (PPCs) in elderly patients undergoing hip fracture surgery.Methods:PubMed, Embase, Cochrance Library, CBM, CNKI and Wanfang Databases were searched.Search strategy was subject word combined with random word.The case control and cohort studies involving the influencing factors for PPCs in elderly patients undergoing hip fracture from January 2000 to November 2020 were retrieved.PPCs included pneumonia, aspiration pneumonitis, pulmonary infections, respiratory failure, atelectasis and pleural effusion, etc.The quality of literatures was assessed by using Newcastle-Ottawa Scale, data were extracted and analysis was performed using RevMan5.3 software.Results:A total of 18 studies were finally enrolled, including 15 case control studies and 3 cohort studies.The results of analysis showed that the advanced age, male, history of smoking, low body mass index, history of cerebrovascular accident, history of chronic obstructive pulmonary disease, history of diabetes, preoperative albumin concentration<35 g/L, preoperative hemoglobin concentration<120 g/L, American Society of Anesthesiologists Physical Status classification≥Ⅲ, operation delay≥48 h and general anesthesia were the risk factors for PPCs ( P<0.05). Conclusions:There are numerous risk factors for PPCs in elderly patients undergoing hip fracture surgery, and advanced age, male, history of smoking, low body mass index, history of cerebrovascular accident, history of chronic obstructive pulmonary disease, history of diabetes, preoperative albumin concentration<35 g/L, preoperative hemoglobin concentration<120 g/L, American Society of Anesthesiologists Physical Status classification≥Ⅲ, operation delay≥48 h and general anesthesia are the risk factors for PPCs in elderly patients undergoing hip fracture surgery.

Objective:To investigate the effects of applying Stent Boost Subtract (SBS) technique during percutaneous coronary intervention in patients with complex coronary lesions.Methods:200 patients with coronary artery disease (CAD) who hospitalized in the department of cardiology in Guangzhou First People′s Hospital from June 2018 to June 2020 were enrolled. The coronary lesions of all patients were corresponding to B2 or C type suggested by American Heart Association (AHA)/American College of Cardiology (ACC) according to coronary angiography and treated with percutaneous coronary intervention (PCI). Patients were randomly divided into SBS group ( n=82, SBS technique was applied during PCI) and IVUS group [ n=118, intravascular ultrasound (IVUS) was applied during PCI]. After stent implantation, quantitative coronary angiography (QCA) automatic analysis system was used to measure the related parameters of stent diameter (including the minimum, maximum and mean value of stent diameter) and calculate the stent eccentricity index. During PCI, stent eccentricity index, post-stent expansion, poorly positioned stent with open lesions, failure of overlapping stent with long lesions, mean cumulative dose (CD), product of total dose area (DAP), X-ray time, operation time and operation cost of each PCI were recorded in the two groups. Patients were followed up for 18 months after PCI, and the occurrence of adverse cardiovascular events (MACE) was recorded during the follow-up period, and the cumulative survival rate without MACE was compared between the two groups. Results:There were no statistically significant differences between the two groups in stent eccentricity index, proportion of guided stent expansion, proportion of poorly positioned stent with open lesions, proportion of stent failure to overlap, with statistically significant difference[(0.12±0.04) vs (0.10±0.03); 80.49% vs 85.49%; 2.44% vs 2.54%; 1.22% vs 2.54%, all P>0.05]. There were no significant differences in CD, X-ray time and DAP in SBS group compared with IVUS group [(1 394.18±42.29)Gy/cm 2 vs (1 391.82±45.06)Gy/cm 2; (18.79±3.01)min vs (18.95±3.12)min, (100.24±5.70)Gy/cm 2 vs (99.47±5.93)Gy/cm 2; all P>0.05]. The operation time in SBS group was shorter than that in IVUS group [(70.91±6.51)min vs (73.89±8.95)min, P<0.05], and the operation cost was less than that in IVUS group [(2.98±0.86)ten thousand yuan vs (3.85±0.81)ten thousand yuan, P<0.05]. After 18 months of follow-up after PCI, Kaplan-Meier survival analysis showed that there was no significant difference in MACE event-free survival between SBS group and IVUS group (91.46% vs 94.07%, Log Rank=0.480, P=0.489). Conclusions:SBS is a kind of convenient and effective technique in guiding PCI in patients with complex coronary lesions without increasing operation time and radiation dose, which can achieve the same effect as IVUS guidance.

Objective:To explore the correlation between middle cerebral artery (MCA) pulsatility index (PI) and total magnetic resonance imaging (MRI) burden in elderly patients with cerebral small vessel diseases (CSVD).Methods:A total of 203 CSVD inpatients aged 60 years and above who were hospitalized in the Department of Neurology of Hebei General Hospital from March 2017 to December 2020 were enrolled. The clinical data, transcranial Doppler ultrasound parameters and brain MRI data were collected. According to the total burden score, the patients were divided into low burden group (0-1 point) and high burden group (2-4 points). Univariate and multivariate Logistic regression analysis was used to analyze the correlation between MCA PI and total MRI burden in the elderly patients with CSVD. Subsequently, the receiver operating characteristic curve was used to evaluate the value of MCA PI for predicting the high MRI burden of CSVD in the elderly.Results:Hypertension ( OR=2.569, 95% CI 1.068-6.182, P=0.035), systolic blood pressure ( OR=1.033, 95% CI 1.006-1.061, P=0.016), creatinine ( OR=1.044, 95% CI 1.009-1.079, P=0.013) and MCA PI ( OR=1.125, 95% CI 1.087-1.166, P<0.001) were independently correlated with the increasing total MRI burden in the elderly patients with CSVD. Spearman rank correlation analysis revealed that there was strong and positive correlation between MCA PI and high MRI burden in the elderly patients with CSVD ( r=0.65, P<0.001). The analysis showed that when the cut-off for MCA PI was 1.11, it could identify high MRI burden of CSVD in the elderly. The area under the curve was 0.908 (95% CI 0.864-0.953, P<0.001). The sensitivity and specificity were 0.852 and 0.880, respectively. The positive predictive value was 92.38%, and the negative predictive value was 77.70%. Conclusion:The MCA PI is positively correlated with total MRI burden in the elderly patients with CSVD, and has a higher value in predicting the total MRI burden in the elderly CSVD patients, which probably bring brighter prospects for its clinical application.

White matter hyperintensity refers to spots or patches changes of white matter in the paraventricular or subcortical regions, which is closely related to the occurrence of cognitive impairment and dementia. This article reviews the neuroimaging examination and imaging evaluation scale of white matter hyperintensity.