Objective: To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability. Methods: A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels. Results: Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment. Conclusion TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Objective : To investigate the role and mechanism of bone formation caused by the ratio of advanced platelet-rich fibrin (A-PRF) and β-tricalcium phosphate (β-TCP) in rabbit femur defect model, which provides a new idea for clinical treatment of bone defect. Methods : Twenty-four New Zealand white rabbits were divided into model group, 1∶1 complex group (A-PRF∶β-TCP=1∶1), 2∶1 complex group (A-PRF∶β- TCP=2∶1) and 4∶1 complex group (A-PRF∶β- TCP=4∶1）, with 6 rabbits in each group. Femoral defect models were constructed in each group. In the composite group, the bone defect was filled with composite material, while in the model group, no material was filled. After 8 weeks, the animals were euthanized and specimens were collected. Bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.SP) and trabecular number (Tb.N) in femoral defect tissue were measured by micro-CT and photographed. Hematoxylin - eosin staining was used to detect the pathological changes of new bone tissue. The morphological changes of the new bone tissue were observed by scanning electron microscopy. Determination of phospho-mitogen activated protein kinase p38 (p-p38MAPK), CCAAT/enhancer binding protein homologous protein (CHOP) and phospho-cysteine aspartic protease-3 (p-Caspase3) in newborn femur by ELISA. The mRNA expressions of osteoprotegerin (OPG), bone morphogenetic protein-2 (BMP-2), receptor activator of nuclear factor kappa-B ligand (RANKL) and p38MAPK were detected by real-time quantitative PCR. The expression of OPG, BMP-2, RANKL, p-p38MAPK and p-Caspase3 protein in the new bone tissue was observed by immunohistochemistry. Results : In the model group, bone formation in the femoral defect area was slow and osteogenic quality was poor. Compared with the model group, the bone formation and neocapillaries of femoral defect area in the complex group was good, BMD, BV.TV, Tb.Th, Tb.N were increased, and Tb.Sp were decreased, the expressions of p-p38MAPK, CHOP and p-Caspase3 were decreased, and the mRNA and protein expressions of OPG and BMP-2 were increased. The mRNA expression of RANKL and p38MAPK was decreased. Apoptosis in new bone tissue of each group showed the lowest apoptosis rate in samples of the 2∶1 complex group (P<0.05); A-PRF: β-TCP=2∶1 ratio has the best osteogenic effect. Conclusion The complex composed of A-PRF and β-TCP can promote the expression of OPG, inhibit the expression of RANKL and phosphorylation of p38MAPK, reduce the apoptosis of new bone tissue cells, and promote osteogenic differentiation.

Objective The risk prediction factors of patients with chronic kidney disease (CKD) complicated with pulmonary tuberculosis were analyzed, and the risk prediction model was constructed to provide theoretical basis for the prevention and treatment of pulmonary tuberculosis in patients with CKD. Methods Stratified sampling was used to randomly select 289 patients with CKD admitted to our hospital as the investigation objects. According to whether patients complicated with tuberculosis, they were divided into experimental group (n=65, CKD complicated with tuberculosis) and control group (n=224, CKD). Univariate analysis and logistic regression were used to analyze the influencing factors of pulmonary tuberculosis in PATIENTS with CKD. According to the regression results, the risk prediction model of pulmonary tuberculosis in CKD patients was established, and the ROC curve was used to predict the efficacy of the model. Results Among 289 patients with CKD, 65 cases (22.49%) had pulmonary tuberculosis. Chest X-ray showed 54 cases of infiltrating pulmonary tuberculosis, 5 cases of voiding pulmonary tuberculosis, 4 cases of caseous pneumonia and 2 cases of tuberculous pleurisy. The main clinical manifestations of CKD complicated with pulmonary tuberculosis were low fever, poor appetite and fatigue in 36 cases, cough and expectoration in 18 cases, high fever in 9 cases and pleural effusion in 2 cases. Mycobacterium tuberculosis culture was positive in 23 cases (35.38%). There were no significant differences in age, CKD stage, past tuberculosis history, low immunity, malnutrition, dialysis treatment, anemia and hypoproteinemia between 2 groups (P<0.05).Multivariate logistic regression analysis showed that low immunity, malnutrition and dialysis treatment were independent risk factors for pulmonary tuberculosis in CKD (P<0.05). Risk of tuberculosis in patients with CKD prediction model for P = 1/(1+e^{-(0.496+0.839×(low immunity)+ 0.892×(malnutrition)+ 1.247×(dialysis)}]; ROC curve was used to analyze the predictive efficacy of the regression model. The results showed that the AUC of pulmonary tuberculosis predicted by the risk prediction model was 0.779, 95%Cl(0.668-0.889) for CKD patients. Conclusion The risk of tuberculosis in CKD is higher,low immunity, malnutrition, dialysis treatment of CKD patients is high risk for tuberculosis, according to the specific situation of the patients, take targeted measures to prevention, can reduce the risk of tuberculosis in patients with CKD.

Objective To analyze the clinical situation and related factors of subclinical hypothyroidism in patients with abnormal glucose metabolism, and to provide theoretical basis for the prevention and treatment of subclinical hypothyroidism in patients with abnormal glucose metabolism. Methods A total of 428 patients with abnormal glucose metabolism who were treated in the Department of Endocrinology of Tianmen First People&#39;s Hospital from March 2018 to March 2020 were selected, and serum FT3, FT4 and TSH levels were determined by automatic immune analysis system. Automatic analyzer was used to measure the levels of FBG, HbA1c, TC, TG, LDL-C and UA. A self-made questionnaire was used to investigate the basic information of all subjects, including gender, age, abnormal course of glucose metabolism, BMI and blood pressure. The survey method was combined with telephone inquiry and field investigation. Logistic regression was used to analyze the independent risk factors for subhypothyroidism in patients with abnormal glucose metabolism. Results Among 428 patients with abnormal glucose metabolism, 89 patients were accompanied by subclinical hypothyroidism, including 39 males and 43 females, with an average age of (45.12±8.13) years. The prevalence of subhypothyroidism in females was higher than that in males, and the difference was statistically significant (χ²=4.353 , P<0.05). There was no significant difference in serum FT3 and FT4 levels between the two groups (P>0.05). The serum TSH level in experimental group was significantly higher than that in control group (P<0.05). Univariate analysis showed statistically significant differences in age, gender, abnormal course of glucose metabolism, BMI, BMI, FBG, HbA1c, UA, TC, LDL-C and SBP between the two groups (P<0.05). Logistic regression analysis showed that old age, high levels of FBG, TC, SBP and UA were independent risk factors for subclinical hypothyroidism in patients with abnormal glucose metabolism (P<0.05). Conclusion The incidence rate of patients with abnormal glucose metabolism complicated with subclinical hypothyroidism is high. The biochemical indexes such as blood glucose, blood lipid, blood pressure and uric acid should be monitored regularly. The early regulation of glucose metabolism disorder is an effective way to prevent and treat subclinical hypothyroidism.

Objective To analyze the distribution characteristics and drug resistance of nontuberculous mycobacteria(NTM),and to provide guidance for the selection of targeted agents in clinical treatment. Methods The clinical data of inpatients in our hospital from April 2019 to February 2021 were collected,the culture and strain identification of non tuberculosis mycobacteria were carried out,the drug sensitivity test of anti tuberculosis drugs was carried out,and the drug resistance of non tuberculosis mycobacteria to first-line anti tuberculosis drugs was analyzed. Results A total of 1 326 strains of mycobacterium were isolated,including 1 154(87.03%)strains of mycobacterium tuberculosis and 172(12.97%)strains of non-mycobacterium tuberculosis.Nine species of nontuberculous mycobacteria were detected,including slow-growing mycobacteria such as Mycobacterium kansasii and Mycobacterium avium-intracellulare complex,belonging to Groups I-III,and fast-growing mycobacteria such as Mycobacterium chelonae and Mycobacterium smegmatis,belonging to Group IV. Among them , Mycobacterium avium-intracellulare complex and Mycobacterium chelonae were dominant,accounting for 26.16%and 36.63%,respectively.Drug susceptibility tests showed that the resistance rate of Mycobacterium avium-intracellulare complex to streptomycin was 100.00%,the drug resistance rate of Mycobacterium chelonae to isoniazid,rifampicin and streptomycin was 100.00%,and the drug resistance rate of Mycobacterium smegmatis and Mycobacterium abscessus to most antibacterial drugs was 100.00%.The resistance rate of major NTM bacteria to clarithromycin was relatively low.There was no statistically significant difference in the susceptibility rates of slow and fast-growing mycobacteria to isoniazid and clarithromycin(P>0.05) ; The susceptibility rates of slow-growing mycobacteria to amikacin,clarithromycin and rifambutin were 62.86%,92.86%and 72.86%,all above 50.00%.The susceptibility rate of the fast-growing mycobacteria to clarithromycin was also more than 50.00%,being 87.25%.The susceptibility rate of slow-growing mycobacteria to other antibiotics was higher than that of fast-growing mycobacteria(P<0.05).The drug resistance of Mycobacterium tuberculosis to first-line anti tuberculosis drugs was significantly lower than that of non Mycobacterium tuberculosis(P<0.05). Conclusion Non-tuberculous mycobacteria have high drug resistance,especially fast-growing mycobacteria,so drug susceptibility tests are of great value in clinical treatment.

[摘 要] 目的：探究miR-323a-3p、四次穿膜蛋白超家族成员1（TM4SF1）在NSCLC组织和细胞中的表达及两者间的靶向调控关系，观察两者表达对A549细胞增殖、迁移、侵袭和裸鼠移植瘤生长的影响。方法：收集2014年1月至12月间青海省人民医院手术切除的20例NSCLC组织及其相应的癌旁组织，qPCR和WB法检测癌组织中miR-323a-3p、TM4SF1 mRNA 和TM4SF1蛋白的表达。向A549细胞转染miR-323a-3p mimic，采用MTT法、Transwell法、WB法检测miR-323a-3p过表达对细胞的增殖、迁移和侵袭以及TM4SF1、细胞周期蛋白D1（cyclin D1）、p21、MMP-2、MMP-9蛋白表达的影响。采用生物信息学预测工具StarBase和双荧光素酶报告基因实验分析miR-323a-3p与TM4SF1靶向关系。将si-TM4SF1转染至A549细胞，以及分别将miR-323a-3p mimic与pcDNA或pcDNA-TM4SF1共转染A549细胞，评估细胞增殖、迁移和侵袭能力的变化；同时建立各组细胞的BALB/c裸鼠移植瘤模型，在14、21和28 d时测量并计算移植瘤体积。结果：与癌旁组织相比，NSCLC组织中miR-323a-3p表达水平明显下调，TM4SF1 mRNA和蛋白表达水平显著上调（均P<0.01）。miR-323a-3p过表达或抑制TM4SF1表达都会降低A549 细胞的增殖、迁移、侵袭能力及cyclin D1、MMP-2、MMP-9蛋白表达而促进p21蛋白表达，并且抑制A549细胞裸鼠移植瘤的生长（均P<0.01）。生物信息学StarBase工具预测和双荧光素酶基因报告实验结果显示miR-323a-3p能够靶向结合TM4SF1基因并调控 TM4SF1的表达。上调TM4SF1表达后，miR-323a-3p过表达对A549细胞恶性生物学行为及cyclin D1、MMP-2、MMP-9蛋白表达、移植瘤生长的抑制作用均被部分逆转（均P<0.01），对p21蛋白表达的促进作用也被逆转（P<0.01）。结论：miR-323a-3p 通过靶向下调肺癌A549细胞中TM4SF1的表达抑制细胞的增殖、迁移、侵袭和裸鼠移植瘤生长。

Objective: To explore the protective effects of anthrahydroquinone-2,6-disulfonate (AH 2 QDS) on the kidneys of paraquat (PQ) poisoned rats via the apelin-APJ pathway. Methods: Male Sprague Dawley rats were divided into four experimental groups: control, PQ, PQ+sivelestat, and PQ+AH 2 QDS. The PQ+sivelestat group served as the positive control group. The model of poisoning was established via intragastric treatment with a 20% PQ pesticide solution at 200 mg/kg. Two hours after poisoning, the PQ+sivelestat group was treated with sivelestat, while the PQ+AH 2 QDS group was given AH 2 QDS. Six rats were selected from each group on the first, third, and seventh days after poisoning and dissected after anesthesia. The PQ content of the kidneys was measured using the sodium disulfite method. Hematoxylin-eosin staining of renal tissues was performed to detect pathological changes. Apelin expression in the renal tissues was detected using immunofluorescence. Western blotting was used to detect the expression levels of the following proteins in the kidney tissues: IL-6, TNF-α, apelin-APJ (the apelin-Angiotensin receptor), NF-κB p65, caspase-1, caspase-8, glucose-regulated protein 78 (GRP78), and the C/EBP homologous protein (CHOP). In in vitro study, a PQ toxicity model was established using human tubular epithelial cells treated with standard PQ. Twenty-four hours after poisoning, sivelestat and AH 2 QDS were administered. The levels of oxidative stress in human renal tubular epithelial cells were assessed using a reactive oxygen species fluorescence probe. Results: The PQ content in the kidney tissues of the PQ group was higher than that of the PQ+AH 2 QDS group. Hematoxylin-eosin staining showed extensive hemorrhage and congestion in the renal parenchyma of the PQ group. Vacuolar degeneration of the renal tubule epithelial cells, deposition of crescent-like red staining material in renal follicles, infiltration by a few inflammatory cells, and a small number of cast formation were also observed. However, these pathological changes were less severe in the PQ+sivelestat group and the PQ+AH 2 QDS group (P<0.05). On the third day after poisoning, immunofluorescence assay showed that the level of apelin in the renal tissues was significantly higher in the PQ+AH 2 QDS group than in the PQ group. Western blotting analysis results showed that IL-6, TNF-α, NF-κB p65, caspase-1, caspase-8, GRP78, and CHOP protein levels in the PQ group were higher than in the PQ+AH 2 QDS group (P<0.05). The expression of apelin-APJ proteins in the PQ+AH 2 QDS group was higher than in the PQ+sivelestat and PQ groups (P<0.05); this difference was significant on Day 3 and Day 7. The level of oxidative stress in the renal tubular epithelial cells of the PQ+AH 2 QDS group and the PQ+sivelestat group was significantly lower than in the PQ group (P<0.05). Conclusions: This study confirms that AH 2 QDS has a protective effect on PQ-poisoned kidneys and its positive effect is superior to that of sivelestat. The mechanism of the protective effects of AH 2 QDS may be linked to reduction in cellular oxidative stress, PQ content of renal tissue, inflammatory injury, endoplasmic reticulum stress, and apoptosis. AH 2 QDS may play a role in the treatment of PQ poisoning by upregulating the expression of the apelin-APJ.

Objective : To construct a visual literacy evaluation system for pupils by Delphi expert consultation method（Delphi method）. Methods : The visual literacy evaluation indices for pupils were preliminarily constructed through literature review. Twenty experts in relevant fields were invited and the visual literacy evaluation system for pupils was established by two rounds of expert consultation based on Delphi method. Results : The average age of 20 experts was（48.35±5.79）years old,of which 19 experts had senior titles,and 17 experts had master&#39;s degree or above. The enthusiasm of the experts in two rounds of consulting was both 100.00%. The average authoritative coefficient（W）of the experts was 0.88. For the first consulting round,the W of importance scores from experts was 0.91（P<0.01）;the W of operability scores from experts was 0.89（P<0.01）,which indicated satisfactory consistency. For the second consulting round,the W of importance scores from experts was 0.79（P<0.01）;the W of operability scores from experts was 0.77（P<0.01）,which also indicated satisfactory consistency. The finalized visual literacy evaluation system for pupils after two rounds of expert consultation included three primary indices（eye care knowledge literacy,eye care attitude literacy and eye care behavior literacy）,six secondary indices（basic knowledge,eye care behavior,eye care attitude,eye care willingness,bad eye care behavior and eye protection）and 40 tertiary indices（≥5.0 of eyesight among normal people,always wearing glasses due to poor eyesight and so on）. Conclusion The experts who participated in the construction of visual literacy evaluation system for pupils had strong professional representativeness,high enthusiasm,high authority and good coordination,and the evaluation system can be used for evaluating pupils visual literacy.

Objective To analyze the curative effect of nitric oxide (NO) and bosentan on treatment of the interruption of aortic arch (IAA) with ventricular septal defect (VSD) and serious pulmonary hypertension (SPH). Methods Thirty-two children with IAA and VSD combined SPH from January 2015 to May 2017 confirmed by cardiac CT and ultrasound in Children’s Hospital of Hebei Province were enrolled including 17 males and 15 females, aged 1.10-4.30 months (mean, 2.71±0.98 months) and weighing 3.33-6.10 kg (mean, 4.57±0.88 kg). The 32 children were randomly divided into two groups (n=16 in each), a NO group and a bosentan group. All the patients underwent interruption of aortic arch and ventricular septal defect repair. When patients returned to cardiosurgery intensive care unit (CSICU) half an hour later, patients in the NO group inhaled NO 20 ppm for 36 h and those in the bosentan group were given bosentan by nasogastric feeding 15 mg, twice a day. The cardic index, pulmonary/systemic pressure ratio, oxygenation index at 3 h, 6 h, 12 h, 24 h, 36 h after surgery were evaluated, and the differences between the two groups were compared. Results The pulmonary/systemic pressure ratio in the two groups increased at first and then decreased, while oxygenation index in the two groups decreased at first and then increased, and the differences in the same groups atthe adjacent time points were statistically significant (P<0.05). The cardiac index in the two groups decreased at first and then increased, the differences in the same groups at the adjacent time points were statistically significant, except for 6 h and 12 h after surgery in the bosentan group (P>0.05). At postoperative 6 h, 12 h, the oxygenation index in the NO group was significantly higher than that in the bosentan group, and the pulmonary/systemic pressure ratio in the NO group was less than that in the bosentan group (P<0.01). The cardiac index in the NO group was higher than that of the bosentan group after 6 h, 12 h, 24 h of operation, which were statistically significant (P<0.05), and the cardic index of children in the NO group was greatly higher than that in the bosentan group after 12 h of surgery (P<0.01); at the same time point, the corresponding indexes were not statistically significant between the two groups (P>0.05). Conclusion NO inhalation in the treatment of IAA with VSD and SPH in children with early postoperative SPH is better than the bosentan, but in the late postoperative period, the effect is similar.