Objective:To investigate the effects of long-term exposure to chrysotile and crocidolite on miRNAs and epithelial mesenchymal transformation (EMT) -related gene expression in human pleural mesothelial cells.Methods:In November 2020, fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of EMT-related genes in human pleural mesothelioma cells (NCl-H2052 cells, NCl-H2452 cells) and human normal mesothelial cells (Met-5A cells). MiRNAs with abnormal expression in human pleural mesothelioma cells were screened out from the previous miRNA chip data of research group, and target genes of differentially expressed miRNAs were predicted using miRWalk database (http: //mirwalk.umm.uni-heidelberg.de). RT-qPCR was used to verify the abnormal expression of EMT-related miRNAs in cell lines. Met-5A cells were treated with 5μg/cm 2 chrysotile and crocidolite respectively for 48 h a time, once a week and a total of 10 times. Chrysotile group, crocidolite group and control group were set up. And the control group was added with the same volume of PBS. The expression changes of EMT-related genes and abnormal expression miRNAs in each group were detected by RT-qPCR. The differences among the groups were compared by one-way ANOVA, and the differences between the control group and the experimental group were compared by dunnet- t test. Results:Compared with Met-5A cells, the expression levels of Vimentin and Twist genes were increased, and the expression level of E-cadherin genes was decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001). Target genes of miRNAs with abnormal expression in miRNA chip were predicted, and the results showed four abnormally expressed miRNAs associated with EMT and verified the expression of these four miRNAs in the cell lines. Compared with Met-5A cells, the expression level of hsa-miR-155-5p was increased in NCl-H2052 cells and NCl-H2452 cells, the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001), which was consistent with the results of chip analysis. After exposure of Met-5A cells, it was found that compared with the control group, the expression levels of Vimentin and Twist genes, hsa-miR-155-5p, hsa-miR-34b-5p and hsa-miR-34c-5p in the crocidolite group were increased, while the expression level of E-cadherin gene was decreased ( P<0.05). Compared with the control group, the expression levels of Vimentin, Twist and E-cadherin genes in chrysotile group were increased, while the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased ( P<0.05) . Conclusion:Long-term exposure to chrysotile and crocidolite could cause Met-5A cells to produce miRNAs and EMT-related gene expression changes similar to mesothelioma cells.

Objective:To investigate the effects of long-term exposure to chrysotile and crocidolite on miRNAs and epithelial mesenchymal transformation (EMT) -related gene expression in human pleural mesothelial cells.Methods:In November 2020, fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of EMT-related genes in human pleural mesothelioma cells (NCl-H2052 cells, NCl-H2452 cells) and human normal mesothelial cells (Met-5A cells). MiRNAs with abnormal expression in human pleural mesothelioma cells were screened out from the previous miRNA chip data of research group, and target genes of differentially expressed miRNAs were predicted using miRWalk database (http: //mirwalk.umm.uni-heidelberg.de). RT-qPCR was used to verify the abnormal expression of EMT-related miRNAs in cell lines. Met-5A cells were treated with 5μg/cm 2 chrysotile and crocidolite respectively for 48 h a time, once a week and a total of 10 times. Chrysotile group, crocidolite group and control group were set up. And the control group was added with the same volume of PBS. The expression changes of EMT-related genes and abnormal expression miRNAs in each group were detected by RT-qPCR. The differences among the groups were compared by one-way ANOVA, and the differences between the control group and the experimental group were compared by dunnet- t test. Results:Compared with Met-5A cells, the expression levels of Vimentin and Twist genes were increased, and the expression level of E-cadherin genes was decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001). Target genes of miRNAs with abnormal expression in miRNA chip were predicted, and the results showed four abnormally expressed miRNAs associated with EMT and verified the expression of these four miRNAs in the cell lines. Compared with Met-5A cells, the expression level of hsa-miR-155-5p was increased in NCl-H2052 cells and NCl-H2452 cells, the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001), which was consistent with the results of chip analysis. After exposure of Met-5A cells, it was found that compared with the control group, the expression levels of Vimentin and Twist genes, hsa-miR-155-5p, hsa-miR-34b-5p and hsa-miR-34c-5p in the crocidolite group were increased, while the expression level of E-cadherin gene was decreased ( P<0.05). Compared with the control group, the expression levels of Vimentin, Twist and E-cadherin genes in chrysotile group were increased, while the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased ( P<0.05) . Conclusion:Long-term exposure to chrysotile and crocidolite could cause Met-5A cells to produce miRNAs and EMT-related gene expression changes similar to mesothelioma cells.

Objective:To demonstrate the three-dimensional structures of the limbic system and its fiber connections through fiber dissection,and to provide reference for relevant professionals to master the anatomy of the limbic system.Methods:Ten cerebral hemispheres were treated and dissected according to Kelinger method,and the limbic system and its fiber connections were displayed.Results:The limbic system was arranged around the thalamus and corpus cal-losum in a double-layer concentric circle structure.The outer layer structures mainly consisted of the cingulate gyrus and the parahippocampal gyrus,while the inner layer structures included the amygdala,hippocampus and fornix.The main association fiber of the outer layer is the cingulum,whose superior trunk is mainly located in the cingulate gyrus,and the inferior trunk is mainly located in the parahippocampal gyrus.The fiber structures of the inner layer includes the striae terminalis and ansa peduncularis emanating from the amygdala and the fornix of the hippocampus.Conclusion:Limbic system is an important connection structure between telencephalon and diencephalon,and its anatomical struc-ture is complex.Fiber dissection method can effectively demonstrate the complex spatial structure of limbic system,which is of great benefit to relevant professionals to understand its three-dimensional structure.

Objective:To demonstrate the three-dimensional structures of the limbic system and its fiber connections through fiber dissection,and to provide reference for relevant professionals to master the anatomy of the limbic system.Methods:Ten cerebral hemispheres were treated and dissected according to Kelinger method,and the limbic system and its fiber connections were displayed.Results:The limbic system was arranged around the thalamus and corpus cal-losum in a double-layer concentric circle structure.The outer layer structures mainly consisted of the cingulate gyrus and the parahippocampal gyrus,while the inner layer structures included the amygdala,hippocampus and fornix.The main association fiber of the outer layer is the cingulum,whose superior trunk is mainly located in the cingulate gyrus,and the inferior trunk is mainly located in the parahippocampal gyrus.The fiber structures of the inner layer includes the striae terminalis and ansa peduncularis emanating from the amygdala and the fornix of the hippocampus.Conclusion:Limbic system is an important connection structure between telencephalon and diencephalon,and its anatomical struc-ture is complex.Fiber dissection method can effectively demonstrate the complex spatial structure of limbic system,which is of great benefit to relevant professionals to understand its three-dimensional structure.

Objective: To investigate craniofacial differences in individuals with hypodontia and explore the relationship between craniofacial features and the number of congenitally missing teeth. Methods: A cross-sectional study was conducted among 261 Chinese patients (males, 124; females, 137; age, 7–24 years), divided into four groups (without hypodontia: no teeth missing, mild: one or two missing teeth, moderate: three to five missing teeth, severe: six or more missing teeth) according to the number of congenitally missing teeth. Differences in cephalometric measurements among the groups were analyzed. Further, multivariate linear regression and smooth curve fitting were performed to evaluate the relationship between the number of congenitally missing teeth and the cephalometric measurements. Results: In patients with hypodontia, SNA, NA-AP, FH-NA, ANB, Wits, ANS-Me/N-Me, GoGn-SN, UL-EP, and LL-EP significantly decreased, while Pog-NB, AB-NP, N-ANS, and S-Go/N-Me significantly increased. In multivariate linear regression analysis, SNB, Pog-NB, and S-Go/N-Me were positively related to the number of congenitally missing teeth. In contrast, NA-AP, FH-NA, ANB, Wits, N-Me, ANS-Me, ANS-Me/N-Me, GoGn-SN, SGn-FH (Y-axis), UL-EP, and LL-EP were negatively related, with absolute values of regression coefficients ranging from 0.147 to 0.357. Further, NA-AP, Pog-NB, S-Go/N-Me, and GoGn-SN showed the same tendency in both sexes, whereas UL-EP and LL-EP were different. Conclusions Compared with controls, patients with hypodontia tend toward a Class III skeletal relationship, reduced lower anterior face height, flatter mandibular plane, and more retrusive lips. The number of congenitally missing teeth had a greater effect on certain characteristics of craniofacial morphology in males than in females.

Traditional Chinese medicine(TCM)plays a unique role in preventive treatment of disease,treatment of chronic disease and recovery.Integrating modern science and technology and methods to carry out integrated and innovative research will help to bring the value of traditional Chinese medicine into full play.In recent years,with the rapid development of digital therapeutics such as artificial intelligence and wearable devices,digital therapeutics have emerged.Under the guidance of TCM theory,TCM integrates digital therapeutics technology methods,personalized treatment of patients through advanced program-driven software or combined with hardware devices,and improves patient compliance,gives full play to the advantages of TCM.This paper uses CiteSpace to visualize the research hotspots of digital therapeutics,combined with the literature and the website data of the Food and Drug Administration and the National Medical Products Administration,to sort out the application of digital therapeutics in the prevention,treatment and recovery of diseases,analyze the key technologies and core theories in the current application of digital therapeutics.And taking TCM pentatotherapy,kinesiatrics and aromatherapy as examples,this paper discusses the application forms of digital therapeutics under the guidance of TCM theory,and provides a preliminary paradigm for TCM digital therapeutics.

Drug repurposing or repositioning has been well-known to refer to the therapeutic applications of a drug for another indication other than it was originally approved for. Repurposing non-oncology small-molecule drugs has been increasingly becoming an attractive approach to improve cancer therapy, with potentially lower overall costs and shorter timelines. Several non-oncology drugs approved by FDA have been recently reported to treat different types of human cancers, with the aid of some new emerging technologies, such as omics sequencing and artificial intelligence to overcome the bottleneck of drug repurposing. Therefore, in this review, we focus on summarizing the therapeutic potential of non-oncology drugs, including cardiovascular drugs, microbiological drugs, small-molecule antibiotics, anti-viral drugs, anti-inflammatory drugs, anti-neurodegenerative drugs, antipsychotic drugs, antidepressants, and other drugs in human cancers. We also discuss their novel potential targets and relevant signaling pathways of these old non-oncology drugs in cancer therapies. Taken together, these inspiring findings will shed new light on repurposing more non-oncology small-molecule drugs with their intricate molecular mechanisms for future cancer drug discovery.

Objective To analyze the basic characteristics, drug features, prescription rules, and drug-symptom relationships of patients in the splenic deficiency and impairment stage, by data mining of medical records under the New Theory on Spleen Dampness Syndrome (Pi Dan Xin Lun, 《脾瘅新论》). Methods Medical records listed in the “New Theory on Spleen Dampness Syndrome – Understanding and Treatment of Metabolic Syndrome from the Perspective of Traditional Chinese Medicine”, and which were diagnosed with the spleen dampness syndrome at the splenic deficiency and impairment stage, during January 2004 and December 2016 were selected. These patients’ data, including basic information, clinical symptoms, laboratory examination results, traditional Chinese medicine (TCM) and western medicine diagnoses, treatment methods, prescriptions, etc., were collected. The collected data were subsequently compiled into a medical record database using the Epidata 3.1 data management software, followed by the use of Apriori algorithm provided in the SPSS Modeler 14.2 statistical software to investigate the association rules between drug-drug, drug-symptom, and drug-western medicine indices. Results (i) A total of 51 medical records were included, involving 17 types of syndromes. Among them, the top three with frequency ≥ 3 included “Phlegm and blood stasis, and thoracic obstruction” “Deficiency-weakness of the spleen Qi, and static blood blocking collaterals”, and “Deficiency-weakness of the spleen Qi, and static blood blocking collaterals”. Alternatively, of the 14 treatment methods, the top three treatments with frequency of ≥ 3 included “Activating Yang and eliminating turbidity, and removing phlegm and dredging channel blockage” “Strengthening the spleen and benefiting Qi, and eliminating phlegm to activate the channels”, and “Warming Yang and benefiting Qi, and expelling cold to remove obstructions”. Among the 15 prescriptions, the top three used with frequency ≥ 3 included Huangqi Guizhi Wuwu Tang (黄芪桂枝五物汤), Gualou Xiebai Banxia Tang (瓜蒌薤白半夏汤), and Ganjiang Huangqin Huanglian Renshen Tang (干姜黄芩黄连人参汤). Lastly, of the 83 drugs used for a total of 476 times, those with frequency ≥ 15 included Huanglian (Coptidis Rhizoma), Huangqi (Astragali Radix), Jiudahuang (Wine-processed Rhei Radix et Rhizoma), Jixueteng (Spatholobi Caulis), Shengjiang (Zingiberis Rhizoma Recens), Huangqin (Scutellariae Radix), and Guizhi (Cinnamomi Ramulus). (ii) For the drug-drug associations, under the criteria of support ≥ 15% and confidence = 100%, seven second-order association rules, seven third-order rules, and six fourth-order roles were identified. The top-ranking rule of each was “Huangqin (Scutellariae Radix) → Huanglian (Coptidis Rhizoma)” “Ganjiang (Zingiberis Rhizoma) + Huangqin (Scutellariae Radix) → Huanglian (Coptidis Rhizoma)”, and “Baishao (Paeoniae Radix Alba) + Guizhi (Cinnamomi Ramulus) + Jixueteng (Spatholobi Caulis) → Huangqin (Scutellariae Radix)”, respectively. Alternatively, the drug-symptom associations were analyzed under the criteria of support ≥ 5% and confidence = 100%, which derived eight second-order association rules, 31 third-order rules, and 30 fourth-order rules. The top-ranking association rule of each order was “Huangqi (Astragali Radix) → Limb edema” “Guizhi (Cinnamomi Ramulus) + Jixueteng (Spatholobi Caulis) → Limb numbness and pain”, and “Guizhi (Cinnamomi Ramulus) + Jixueteng (Spatholobi Caulis) + Huangqi (Astragali Radix) → Limb numbness and pain”, respectively. Similarly, the drug-western medicine index associations were investigated under the criteria of support ≥ 5% and confidence = 100%, and five second-order association rules, 16 third-order rules, and 16 fourth-order rules were identified. In this category, the top-ranking association rule of each order was “Qinpi (Fraxini Cortex) → Uric acid” “Huanglian (Coptidis Rhizoma) + Ganjiang (Zingiberis Rhizoma) → Glycated hemoglobin”, and “Huanglian (Coptidis Rhizoma) + Ganjiang (Zingiberis Rhizoma) + Huangqin (Scutellariae Radix) → Glycated hemoglobin”, respectively. Conclusion Through association rule mining, this study objectively and quantitatively demonstrated the drug-drug, drug-symptom, and drug-physicochemical index associations of patients with the spleen dampness syndrome at the splenic deficiency and impairment stage treated by Academician TONG Xiaolin. The results indicated that treatment for these patients adopted the “state-target” syndrome differentiation method. The drug combination was characterized by “small prescriptions”, targeting both the patient’s symptoms and signs (syndrome target) and western medicine indices (treatment target). This study could provide references for future research on the academic thoughts and medical experience of Academician TONG Xiaolin.

Objective To investigate the effect ofdiammonium glycyrrhizinate on neurovascular units in rats after cerebral ischemia reperfusion (IR) injury.Methods Two hundred and forty health SD rats were randomly assigned into normal control group (n=30),sham-operated group (n=30),IR group (n=90) and diammonium glycyrrhizinate group (DG,n=90).The rats in the IR group and DG group were divided into 2,6 and 12 h subgroups after modeling,respectively (n=30).The rats in the IR group and DG group were induced middle cerebral artery occlusion (MCAO) models,and after the models were successfully established,9.11 mL DG sodium chloride injection was given to DG group,while equal saline to normal group,sham-operated group and IR group via the tail vein.The brain tissues of each group were harvested 2,6 and 12 h,resperctively,after modeling.The infraction rate was measured by TTC staining;immunohistochemistry was employed to detect the expresions of Claudin-5 and vessel endothelium (VE)-Cadherin;Western blotting was used to detect the protein expression levels of Rac-1 and Claudin-5.Results The DG group had signficantly lower infarction rate than IR group 2,6 and 12 h after modeling (P＜0.05).The Claudin-5 expression rates in the 6 h and 12 h DG subgroups were signficantly higher than those in the 6 h and 12 h IR subgroups (P＜0.05).The VE-Cadherin expression rates in the DG group were significantly higher than that in IR group at 2,6 and 12 h after modeling (P＜0.05).Samely,the Claudin-5 relative quantity in DG group was significantly higher than that in IR group at 2,6 and 12 h after modeling (P＜0.05).The Rac-1 quantity in DG group was only statistically higher than IR group at 2 h after modeling (P＜0.05).Conclusion The DG can upregulate the Rac-1,VE-Cadherin and Claduin-5 expressions in neurovascutar units,and partly protect neurovascular units after cere bral acute IR injury.

Objective To discuss the feasibility of neuroendoscopic third ventriculostomy for chronic posttraumatic hydrocephalus (PTH).Methods Nineteen cases of chronic PTH treated with neuroendoscopic third ventriculostomy between October 2010 and October 2014 were analyzed retrospectively.There were 13 males and 6 females, aged 11-57 years (mean, 36.3 years).Trauma resulted from traffic accidents in 14 cases, falls in 4 cases and blunt object hitting in 1 case.Of the 19 cases analyzed, 5 had Glasgow Coma Scale (GCS) score of 13-15, 5 had score of 9-12 and 9 had score of 5-8 at admission.Results of operation were assessed with the Canada multicenter evaluation criteria.Prognosis was analyzed with the Glasgow Outcome Scale (GOS).Results All cases were followed up for mean 13.6 months (range, 6-26 months).Improvement of symptoms was achieved in 17 cases, but was not seen in 2 cases.Of the 2 cases, one required ventriculoperitoneal shunt two weeks after ineffective ventriculostomy, and one required second ventriculostomy one month after the presence of stoma blockage.No serious complications occurred.At follow-up, 9 cases had GOS score of 5, 8 cases had score of 4 and 2 cases had score of 3.Conclusions Neuroendoscopic third ventriculostomy is in line with the physical characteristics in cerebrospinal fluid circulation, which implies no shunt implantation, less operative trauma and less complications.The procedure is an effective approach for chronic PTH.