BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

Objective:To investigate the expression of von Hippel-Lindau (VHL), vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1α (HIF-1α) in endolymphatic sac tumor (ELST) and its clinical significance, and to analyze its association with VHL gene mutation.Methods:Twenty-four cases of ELST, which were surgically resected and diagnosed by pathological examination in Beijing Tongren Hospital Affiliated to Capital Medical University, Beijing, China during 2012-2020, were recruited as the ELST group, and 24 cases of otitis media diagnosed in the same hospital were selected as the control group. The expression of VHL, VEGF, and HIF-1α was assessed using EnVision immunohistochemical staining and compared between the ELST and control groups. Sanger sequencing was performed to detect the VHL mutation status in 24 ELSTs. The correlations among VHL, VEGF and HIF-1α expression were analyzed. The associations of VHL, VEGF and HIF-1α expression with age of onset, gender, tumor size, bone invasion and clinical stage in ELST were also analyzed.Results:The expression rate of VHL in the ELST group was significantly lower than that in the control group ( P<0.05), but the expression rates of VEGF and HIF-1α in the ELST group were significantly higher than those in the control group ( P<0.05). VHL expression was inversely correlated with VEGF and HIF-1α expression. The expression of VEGF and HIF-1α was associated with bone invasion and clinical stage ( P<0.05), but the expression of VHL, VEGF and HIF-1α had no significant associations with the age of onset, gender, or tumor size of ELST ( P>0.05). Conclusions:The expression of VHL is decreased while that of VEGF and HIF-1α increased in ELST. Expression of VHL is inversely correlated with that of VEGF and HIF-1α. The expression of VEGF and HIF-1α is correlated with bone invasion and clinical stage. Thus, VEGF and HIF-1α may be therapeutic targets of ELST.

Objective:To analyze the research trends of benzene poisoning in recent years, to highlight the relevant research hotspots and prospects through bibliometrics.Methods:In August 2020, used "benzene" methylbenzere as key words, publications were retrieved from the PubMed, the Web of Science Core Collection database, SinoMed and CNKI before August 31, 2020. Describing the publication time, the number of the published research, publication countries, research institutions and journals. VOSviewer 1.6.10 were used to evaluate and co-occurrence analyze the keywords of the researches.Results:There were 510 English literatures about benzene poisoning. 43 countries had published relevant literatures. The United States had the largest number of articles (43.92%, 224/510) in this field, and China ranked second (11.57%, 59/510) . Relevant results had been published in 257 English journals. The most widely published journal was Environmental Health Perspectives (4.90%, 25/510) . There were 536 Chinese literatures. 368 research institutions had published relevant literatures. The first one was Shenzhen Occupational Disease Prevention and Control Hospital (3.36%, 18/536) , followed by the Chinese Center for Disease Control and Prevention (3.17%, 17/536) . A total of 170 journals had published relevant research results. The most frequently published journal is Chinese Journal of Industrial Hygiene and Occupational Diseases (12.13%, 65/536) . Key words co-occurrence analysis indicated that the domestic and foreign literatures paid more attention to occupational exposure, blood toxicity, myelosuppression, genetic polymorphism and so on.Conclusion:Domestic and foreign researches on benzene poisoning are mostly in China and the United States. In recent years, the research has gradually focused on the related mechanism of benzene poisoning, such as blood toxicity, carcinogenicity, genotoxicity and immunotoxicity.

Objective:To analyze the research trends of benzene poisoning in recent years, to highlight the relevant research hotspots and prospects through bibliometrics.Methods:In August 2020, used "benzene" methylbenzere as key words, publications were retrieved from the PubMed, the Web of Science Core Collection database, SinoMed and CNKI before August 31, 2020. Describing the publication time, the number of the published research, publication countries, research institutions and journals. VOSviewer 1.6.10 were used to evaluate and co-occurrence analyze the keywords of the researches.Results:There were 510 English literatures about benzene poisoning. 43 countries had published relevant literatures. The United States had the largest number of articles (43.92%, 224/510) in this field, and China ranked second (11.57%, 59/510) . Relevant results had been published in 257 English journals. The most widely published journal was Environmental Health Perspectives (4.90%, 25/510) . There were 536 Chinese literatures. 368 research institutions had published relevant literatures. The first one was Shenzhen Occupational Disease Prevention and Control Hospital (3.36%, 18/536) , followed by the Chinese Center for Disease Control and Prevention (3.17%, 17/536) . A total of 170 journals had published relevant research results. The most frequently published journal is Chinese Journal of Industrial Hygiene and Occupational Diseases (12.13%, 65/536) . Key words co-occurrence analysis indicated that the domestic and foreign literatures paid more attention to occupational exposure, blood toxicity, myelosuppression, genetic polymorphism and so on.Conclusion:Domestic and foreign researches on benzene poisoning are mostly in China and the United States. In recent years, the research has gradually focused on the related mechanism of benzene poisoning, such as blood toxicity, carcinogenicity, genotoxicity and immunotoxicity.

Objective: To explore the application values of prenatal ultrasound, vascular cast in the diagnosis of fetal aortic arch and its branches anomalies and to analyze the genetic characteristics by gene detection. Methods: Twenty-two cases of the vascular cast specimens of the fetal aortic arch and its branches anomalies were analyzed and studied by comparing with their prenatal ultrasonography. Then the characteristics of each type of fetal aortic arch and its branches anomalies, the missed diagnosis and misdiagnosis were summarized and the results of their gene detection were also analyzed. Results: The 22 cases of fetal aortic arch and its branches anomalies were as follows: 2 cases of double aortic arch showed the ascending aorta was divided into two branches after converging as the descending aorta. Three cases were left aortic arch with aberrant right subclavian artery. Twelve cases were right aortic arch: 8 cases were right aortic arch with mirror-image branching, 3 cases were right aortic arch with aberrant left subclavian artery, 1 case was right aortic arch with isolated left subclavian artery. Of the 8 right aortic arch with mirror-image branching, 3 cases of left arterial duct showed the vertical walking between the fusion site of the left innominate artery and the pulmonary artery. Right aortic arch with aberrant left subclavian artery with arterial duct showed "U" shaped vascular ring. Five cases were other types, including 2 cases of the coarctation of aortic arch, 1 case of interrupted aortic arch, 1 case of pulmonary artery sling, and 1 case of abnormal origin of right pulmonary artery. The ultrasonic missed diagnosis were the 6 deformities: 3 cases of arterial duct and 3 cases of aberrant subclavian artery. The ultrasonic misdiagnosis were the 5 deformities: 2 cases of arterial duct location, 1 case of aberrant subclavian artery, 1 case of isolated left subclavian artery, and 1 case of the coarctation of aortic arch. Genetic test results: In the 18 cases of the genetic detection, 2 cases were positive, 1 case was 22q11.2 microdeletion syndrome and 1 case was carrying KMT2D gene variant. Conclusions There are various kinds of fetal aortic arch and its branches anomalies, which are often associated with intracardiac malformations and venous branches variation. And prenatal ultrasound is of great value in diagnosing them.Vascular cast can visually display their characteristic changes, which is helpful to improve the differential diagnosis of the different aortic arch and its branches anomalies. The detailed genetic detection can improve the further understanding of its etiology.

Objective To explore the correlative factors of sudden death in patients suspected with pulmonary thromboembolism (PTE) in emergency room (ER).Methods A retrospective analysis was conducted.The clinical data of 12 patients with sudden death suspected with PTE (sudden death group) in ER of the Air Force General Hospital from January 2011 to June 2014 were analyzed.The non-sudden death group included 35 patients during the same time period who were diagnosed with PTE based on findings of CT pulmonary arteriography (CTPA) and showed no sudden death in ER.Factors,including sex,age,previous operation,tumor,syncope,dyspnea,bilateral or unilateral edema of lower extremity,heart rate (HR),white blood cell count (WBC),D-dimer,arterial partial pressure of oxygen (PaO2),arterial partial pressure of carbon dioxide (PaCO2) and typical clinical manifestation of electrocardiogram (SⅠTⅢQⅢ),were compared between the two groups.The potential predictors of sudden death of PTE were analyzed by logistic regression analysis.Results Young age (years old:51.3±15.5 vs.62.3±14.4),lower PaO2 [mmHg (1 mmHg =0.133 kPa):49.9± 12.3 vs.62.7± 10.2],higher HR (bpm:122.0± 19.5 vs.89.1 ± 18.5) and higher WBC (× 109/L:13.8 ± 6.9 vs.7.2 ± 2.5) were found in sudden death group as compared with those in non-sudden death group (P ＜ 0.05 or P ＜ 0.01).There was no significant differences in D-dimer level and PaCO2 between sudden death group and non-sudden death group [D-dimer (pg/L):986 (891,3 230) vs.2089 (598,3 397),PaCO2 (mmHg):33.0 (28.6,43.4)vs.36.5 (32.9,41.0),both P ＞ 0.05].The syncope,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months,bilateral asymmetrical edema in sudden death group were more than those of the non-sudden death group,and chest pain was less (P ＜ 0.05 or P ＜ 0.01).Difference in gender,dyspnea and typical SⅠTⅢQⅢ in electrocardiogram were not significant between the two groups (all P ＞ 0.05).It was shown by multiple logistic regression analysis that higher HR [odds ratio (OR) =1.124,95％ confidence interval (95％CI) =1.024-1.235,P =0.014] and higher WBC (OR =1.347,95％CI =1.043-1.738,P =0.022) were identified as independent risk factors of sudden death for PTE.Conclusions Gender,dyspnea,typical S Ⅰ TⅢQⅢ in electrocardiogram,PaCO2 and D-dimer seem unrelated to sudden death of patients with PTE.Young age,chest pain,syncope,bilateral asymmetrical edema,antineoplaston treatment or tumor metastasis within 6 months,operation in previous 4 months and low PaO2 were potential predictors of sudden death according to the univariate analysis.Higher WBC and higher HR are independent risk factors of sudden death for PTE patients.

This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage.

Background and purpose:Endoscopic treatment is a promising therapeutic option for superifcial lesions throughout the gastrointestinal tract, this study was aimed to evaluate the efficacy of endoscopic resection (ER) using the new Duette multiband mucosectomy kit (DT-6) on treating esophageal disease. Methods:Since Jun. 2011, ER using DT-6 has been performed on 100 patients in a tertiary medical center. Data from those who have been followed up for over 6 months was analyzed. ER and esophagectomy were compared on treating high grade dysplasia (HGD) lesions and early esophageal cancer. Results:From Jun. 2011 to Jan. 2012, a total of 32 patients with esophageal lesions underwent 34 ER using DT-6 (22 male and 10 female, mean age 59.0 years, range 25 to 83 years). There were (3.4±1.0) specimen resected per operation, and the average greatest diameter was (11.8±2.7)mm. Intraoperative blood loss was (5.45±1.47)mL. The median follow-up period was 8.2 months with a 100%half-year-follow-up rate. Except one pneumothorax occurred during one endoscopic submucosal dissection (ESD), no other complications happened. When Comparing ER and esophagectomy on treating HGD and early esophageal cancer, ER showed advantages in terms of operation time, intraoperative blood loss, hospital stay and complications. Conclusion:ER using DT-6 is safe, simple, minimally invasive and effective for esophageal disease. Prospective study and long follow-up are needed to compare endoscopic resection and esophagectomy for HGD and early esophagus cancer.

Marine actinomyces LYG-1 was isolated from marine mud flats in Lianyungang,China.Strain LYG-1 was identified using the methods of morphology,physiological and ehemotaxonomic characterization and 16S rRNA gene sequence analysis.The results showed that strain LYG-1 was a marine variable species of Streptomyces roseosporus.The fermentation broth of strain LYG-1 exhibited conspicuous antitumor activity against HepG2,MCF-7,HCT116 and MDA-MB-231 cell lines,and the IC50 values were defined by MTT method respectively.

ObjectiveTo explore the effects of brucine on multiple myeloma (MM) and to compare the effects between brucine and bontezomib on MM.MethodsMTT method was used to determine the median inhibitory concentratiom (IC50) of brucine and bortezomib on the MM cell line U266.The supernatant of cultured U266 cell line was added to the culture system for inducing the differentiation of osteoblast cell line MC3T3-E1.After aseptic assay,RT-PCR was used to determine the RNA levels of alkaline phosphatase (ALP),osteocalcine (OC),osteoprotegerin (OPG) and recepter activator of NF-κB ligand (RANKL).Results IC50 of bortezomib on U266 cell line for 48 h was 22.4 nmol/L,and that of strychnine was 0.16 nmol/L.The mRNA levels of ALP,OC and OPG in osteoblast co-intervened by brucine combined with the supernatant of MM cells were higher than those in supernatant of U266 cells,while the level of RANKL mRNA was lower (P ＜0.05).The degree of increasing or reducing was greater than the level of control group intervened only by bortezomib (P ＜0.05).ConclusionThe therapeutic effects of brucine on MM might be carried out through the regulation of osteoclast by osteoblast,and the experiment confirmed that the therapeutic effect of brucine on MM was superior to that of bortezomib.