Diagnostic ultrasound-mediated microbubble cavitation dose-dependently improves diabetic cardiomyopathy through angiogenesis.

Ultrasound-mediated microbubble cavitation (UMMC) induces therapeutic angiogenesis to treat ischemic diseases. This study aimed to investigate whether diagnostic UMMC alleviates diabetic cardiomyopathy (DCM) and, if so, through which mechanisms. DCM model was established by injecting streptozocin into rats to induce hyperglycemia, followed by a high-fat diet. The combined therapy of cation microbubble with low-intensity diagnostic ultrasound (frequency = 4 MHz), with a pulse frequency of 20 Hz and pulse length (PL) of 8, 18, 26, or 36 cycles, was given to rats twice a week for 8 consecutive weeks. Diagnostic UMMC therapy with PL at 8, 18, and 26 cycles, but not 36 cycles, dramatically prevented myocardial fibrosis, improved heart functions, and increased angiogenesis, accompanied by increased levels of PI3K, Akt, and eNOS proteins in the DCM model of rats. In cultured endothelial cells, low-intensity UMMC treatment (PL = 3 cycles, sound pressure level = 50%, mechanical index = 0.82) increased cell viability and activated PI3K-Akt-eNOS signaling. The combination of diagnostic ultrasound with microbubble destruction dose-dependently promoted angiogenesis, thus improving heart function through PI3K-Akt-eNOS signaling in diabetes. Accordingly, diagnostic UMMC therapy should be considered to protect the heart in patients with diabetes.

Exploring the effects of artesunate and fuzheng huayu decoction on mitochondria in the treatment of schistosomiasis liver fibrosis / 中华肝脏病杂志

Objective: To compare the effects of artesunate (Art) and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis. Methods: Eighty C57BL/6 female mice were randomly divided into healthy control group, infection group, Art treatment group and Fuzheng Huayu Decoction treatment group, with 20 mice in each group. Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae. After 6 weeks, praziquantel (300 mg/kg) was used for 2 days to kill the worms. The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day, while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day. After 6 weeks, fresh liver tissues of the four groups were collected. Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A (SDHA) and malate dehydrogenase (MDH2), citrate synthase (CS), ketoglutarate dehydrogenase (OGDH), and target of rapamycin 1 (mTORC1) pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues. The relative expression levels of adenylate activated protein kinase (AMPK) and mitochondrial autophagy pathway kinase (PINK1) were detected. Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate. Two-way ANOVA was used to compare the significance and difference between two sets of samples. Results: Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group. Western blot analysis showed that the infection group (0.82 ± 0.05) had significantly lower relative expression of SDHA protein than the healthy control group (1.00 ± 0.05) (t = 11.23, P = 0.0035), while the Art treatment group (0.73 ± 0.05) had significantly higher relative expression of SDHA protein than the infection group (t = 10.79, P = 0.0073). However, there was no significant change in Fuzheng Huayu Decoction treatment group (0.98±0.05) (t = 1.925, P = 0.1266). The relative expression of p-AMPK protein was significantly higher in the infection group (1.15 ±0.05) than in the healthy control group (0.98 ± 0.07, t = 12.18, P = 0.0029), and the expression of p-AMPK in the Art treatment group (0.50 ± 0.05) was significantly lower than the infection group (t = 11.78, P = 0.0032). The relative protein expression of AMPK was significantly lower in the infection group (0.80 ± 0.05) than in the healthy control group (1.00 ± 0.05, t = 10.53, P = 0.0046). The expression of AMPK was significantly lower in the Art treatment group (0.54 ± 0.05) than in the infection group (T = 13.98, P = 0.0036). The relative expression of p-mTORC1 protein (0.93 ± 0.08) was not significantly different in the infection group than in the healthy control group (t = 2.28, P = 0.065), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1 protein than the infection group (t = 10.58, P = 0.029). The expression of p-mTORC1/ m-TORC1 was not significantly different in the infection group (0.98 ± 0.03) than in the healthy control group (0.97 ± 0.03, t = 0.98, P = 0.085), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1/ m-TORC1 than the infection group (t = 14.58, P = 0. 009). The relative protein expression of PINK1 was significantly lower in the infection group (0.55 ± 0.05) than in the healthy control group (1.00 ± 0.03, t = 13.49, P = 0.0011), while the Art treatment group (1.21 ± 0.05, t = 9.98, P = 0.0046) and Fuzheng Huayu Decoction treatment group (1.31 ±0.35, t = 6.98, P = 0.027) had significantly higher relative protein expression of PINK1 than the infection group. Mitochondrial function tests showed that after adding substrate complex II, the oxygen consumption of the infection group was lower than the healthy control group, while the Art treatment group and the Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. The oxygen consumption was significantly lower after adding the substrate complex III in the infection group than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. Conclusion: Art can alleviate schistosomiasis liver fibrosis by inhibiting AMPK/mTORC1 signaling pathway activity and enhancing mitochondrial oxygen consumption, autophagy and SDHA expression.

Preparation and in vitro evaluation of fused deposition modeling 3D printed compound tablets of captopril and hydrochlorothiazide / 北京大学学报(医学版)

OBJECTIVE@#To explore the feasibility of preparing compound tablets for the treatment of hypertension by fused deposition modeling (FDM) 3D printing technology and to evaluate the quality of the printed compound tablets in vitro.@*METHODS@#Polyvinyl alcohol (PVA) filaments were used as the exci-pient to prepare the shell of tablet. The ellipse-shaped tablets (the length of major axes of ellipse was 20 mm, the length of the minor axes of ellipse was 10 mm, the height of tablet was 5 mm) with two separate compartments were designed and printed using FDM 3D printer. The height of layer was 0.2 mm, and the thickness of roof or floor was 0.6 mm. The thickness of shell was 1.2 mm, and the thickness of the partition wall between the two compartments was 0.6 mm. Two cardiovascular drugs, captopril (CTP) and hydrochlorothiazide (HCT), were selected as model drugs for the printed compound tablet and filled in the two compartments of the tablet, respectively. The microscopic morphology of the tablets was observed by scanning electron microscopy (SEM). The weight variation of the tablets was investigated by electronic scale. The hardness of the tablets was measured by a single-column mechanical test system. The contents of the drugs in the tablets were determined by high performance liquid chromatography (HPLC), and the dissolution apparatus was used to measure the in vitro drug release of the tablets.@*RESULTS@#The prepared FDM 3D printed compound tablets were all in good shape without printing defects. The average weight of the tablets was (644.3±6.55) mg. The content of CTP and HCT was separately (52.3±0.26) mg and (49.6±0.74) mg. A delayed in vitro release profile was observed for CTP and HCT, and the delayed release time for CTP and HCT in vitro was 20 min and 40 min, respectively. The time for 70% of CTP and HCT released was separately 30 min and 60 min.@*CONCLUSION@#CTP and HCT compound tablets were successfully prepared by FDM 3D printing technology, and the printed tablets were of good qualities.

Etiology and Pathogenesis of Hepatolenticular Degeneration Caused by Latent Toxin Blocking Collaterals / 中国实验方剂学杂志

Hepatolenticular degeneration（HLD），also known as Wilson disease （WD）， is a genetic disorder characterized by copper metabolism disorder caused by ATP7B gene mutation. Specifically， due to the ceruloplasmin synthesis disorder induced by gene mutation，copper cannot be excreted through bile，which results in pathological deposition of copper in various organs and damage to organs such as the brain and the liver. The incidence of WD in Chinese is significantly higher than that in the world. Copper chelating agents， such as D-penicillamine and dimercaptosuccinic acid， are used as the main therapeutic agents in western medicine. However， many clinical adverse events limit the application of these drugs. Traditional Chinese medicine （TCM） has its characteristics in the treatment of WD. As confirmed by long-term research on TCM clinical diagnosis and treatment，MD has become TCM dominant disease. In spite of many views about the etiology and pathogenesis of WD，a consensus has not been reached so far. Based on the theory of latent pathogen in TCM and the pathological mechanism of excessive deposition of copper ions in the body，this study proposed that latent toxin is the key etiology of WD，and further elaborated that the latent toxin of WD was inherited from parents and occurred in children and adolescents，which was hidden in the liver and the kidney and damaged the brain. The latent toxin， Yang in nature and dispersing in property， is prone to transform into dampness-heat to block Qi movement and produce phlegm leading to stasis. Furthermore， this study determined latent toxin blocking collaterals as the basic pathogenesis of WD and revealed the complex clinical manifestations of latent toxin blocking collaterals such as liver collaterals，brain collaterals，kidney collaterals，spleen collaterals，stomach collaterals，lung collaterals，heart collaterals， and uterus collaterals. Treatment should follow the basic therapeutic principles of resolving pathogens，removing toxins， and dredging collaterals. This study is expected to provide a theoretical basis for syndrome differentiation and treatment of WD in TCM.

Intervention of Gandou Fumu Decoction on Liver Fibrosis in Mice with Wilson's Disease Through JNK Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo identify the protective effect and possible mechanism of Gandou Fumu decoction （GDFMD） on liver fibrosis in mice with Wilson's disease. MethodA total of 50 homozygous TXJ mice were randomly divided into five groups， with 10 mice in each group. Ten wild-type mice were selected as a normal group. The GDFMD high， medium， and low-dose groups were given 13.92， 6.96， 3.48 g·kg-1 of GDFMD， respectively. The penicillamine group were given 0.1 g·kg-1 of penicillamine. The model group and the normal group were given the same volume of 0.9% sodium chloride solution once a day for 4 consecutive weeks. The enzyme-linked immunosorbent assay （ELISA） method was performed to detect serum superoxide dismutase （SOD） activity and malondialdehyde （MDA） content. Corresponding kits were used to detect the mitochondrial adenine triphosphate （ATP） content and Na+-K+-ATPase activity in liver tissues. Hematoxylin-eosin （HE） and Masson staining were used to observe the pathological morphology of liver tissue， and transmission electron microscope was used to observe ultrastructural changes of liver tissues in mice. Western blot was used to detect the c-Jun N-terminal kinase， the phosphorylated protein， and the expressions of Caspase-3， B cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in c-Jun N-terminal kinase （JNK） signaling pathway. ResultCompared with the normal group， MDA content increased and SOD activity decreased in the model group （P<0.05）. Compared with the model group， SOD activities in the GDFMD high-， medium-， and low-dose groups and the penicillamine group significantly increased （P<0.01）， and MDA content significantly decreased （P<0.05， P<0.01）. Compared with the normal group， ATP content and Na+-K+-ATPase activity significantly decrease in the model group （P<0.05）. Compared with the model group， ATP content and Na+-K+-ATPase activity in the GDFMD medium and high-dose groups and the penicillamine group significantly increased （P<0.05， P<0.01）. The results of the pathological morphology of liver tissue showed that a large number of liver cells degeneration and necrosis， inflammatory cell infiltration， unclear liver lobule structure， and collagen fiber deposition were observed in the model group. Transmission electron microscopy showed that the number of mitochondria in liver tissues significantly reduced， the mitochondria were locally damaged， and the cristae of mitochondria were broken even disappear in the model group. The pathological morphology of liver tissue and mitochondrial structure recovered to varying degrees after medicinal intervention. The results of Western blot suggested that， compared with the normal group， the expression levels of phosphorylation-JNK （p-JNK）， p-JNK/JNK， Caspase-3， and Bax in the liver tissues were up-regulated， while the expression of Bcl-2 was down-regulated in the model group （P<0.05）. The expression levels of p-JNK， p-JNK/JNK， Caspase-3 and Bax were down-regulated and the expression of Bcl-2 was up-regulated in the GDFMD high and medium-dose groups and the penicillamine group （P<0.01）. ConclusionGDFMD can alleviate oxidative stress damage and recover mitochondrial function of TXJ mice with liver fibrosis. The mechanism of GDFMD may be related to regulating the JNK signaling pathway and downstream factors and inhibiting cell apoptosis.

Automated Framework and Visualization-based Analysis of Concept Evolution of Spleen and Stomach Diseases and Syndromes in Traditional Chinese Medicine / 中国实验方剂学杂志

ObjectiveTo collect and screen records concerning the spleen and stomach diseases and syndromes in ancient books of traditional Chinese medicine （TCM） using an automated framework and to systematically explore the concept evolution of spleen and stomach diseases and syndromes using the visualization method. MethodA total of 1 224 kinds of ancient book data in the Ancient Books of Traditional Chinese Medicine Database （V2.1） were analyzed using the automated testing tool Selenium WebDriver with the Python 3.8 programming language and the etree library of Lxml for automatic collection and statistics of the "book title" "author" "classification" "dynasty" "completion time"， and "informative abstract". After being checked and collated， the collected data were visually analyzed with Tableau （V2020.1.3） for figuring out the concept evolution of spleen and stomach diseases and syndromes in the past dynasties from the perspectives of symptoms and signs， etiology and pathogenesis， principle-method-recipe-medicinal， and prognosis. ResultA total of 7 203 clauses were automatically collected from 989 ancient books. It was found that in the pre-Qin period， there were few ancient books related to the spleen and stomach diseases and syndromes， and the understandings were confined to the superficial symptoms or signs and the basic etiology and pathogenesis. From the Han to Sui and Tang dynasties， the related concepts gradually increased and the descriptions about the manifestations are more detailed than those in previous dynasties. The etiology， diagnosis， and treatment system of the spleen and stomach diseases and syndromes were further perfected. In the Song， Jin， and Yuan dynasties， such concepts as independent signs，symptoms， as well as nature and location of spleen and stomach diseases and syndromes were enriched. In the Ming and Qing dynasties， a TCM syndrome differentiation and treatment system for spleen and stomach diseases and syndromes was formed， and the related concepts were gradually simplified and unified. ConclusionThe concepts of spleen and stomach diseases and syndromes have undergone an evolution from simplicity to complexity and then back to simplicity. There are numerous ancient books discussing the concepts of spleen and stomach diseases and syndromes， exhibiting a fluctuating yet rising trend with time. The automated framework enables the construction of a lightweight database of spleen and stomach diseases and syndromes. Based on data visualization， the concept evolution of the spleen and stomach diseases and syndromes from ancient times to the present has been efficiently uncovered， which is conducive to tracing the origin and development of spleen and stomach diseases and syndromes in TCM. This has provided reference for related research of spleen and stomach diseases in modern Chinese medicine.

Potential inhibitors for blocking the interaction of the coronavirus SARS-CoV-2 spike protein and its host cell receptor ACE2

The outbreak of SARS-CoV-2 continues to pose a serious threat to human health and social and economic stability. In this study, we established an anti-coronavirus drug screening platform based on the Homogeneous Time Resolved Fluorescence (HTRF) technology and the interaction between the coronavirus S protein and its host receptor ACE2. This platform is a rapid, sensitive, specific, and high throughput system. With this platform, we screened two compound libraries of 2,864 molecules and identified three potential anti-coronavirus compounds: tannic acid (TA), TS-1276 (anthraquinone), and TS-984 (9-Methoxycanthin-6-one). Our in vitro validation experiments indicated that TS-984 strongly inhibits the interaction of the coronavirus S-protein and the human cell ACE2 receptor. This data suggests that TS-984 is a potent blocker of the interaction between the S-protein and ACE2, which might have the potential to be developed into an effective anti-coronavirus drug. SIGNIFICANCEThe ongoing pandemic of COVID-19 caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has made a serious threat to public health worldwide. Given the urgency of the situation, researchers are attempting to repurpose existing drugs for treating COVID-19. In this present study, we screened two compound libraries of 2,864 molecules and identified a potent inhibitor (TS-984) for blocking the coronavirus S-protein and the human cell ACE2 receptor. TS-984 might have the potential to be developed into an effective anti-coronavirus drug for treating COVID-19.

General recommendation for assessment and management on the risk of glucocorticoid-induced osteonecrosis in patients with COVID-19.

BACKGROUND/OBJECTIVE: Coronavirus disease 2019 (COVID-19) is a disaster in human medical history and glucocorticoids remain the most promising therapy. Osteonecrosis is a disease caused by reduced intraosseous blood flow to bones in the joints, which will rapidly induce joint destruction. Approximately one-third patients with severe acute respiratory syndrome (SARS) who received high cumulative doses and long treatment durations of glucocorticoids occurred osteonecrosis. Considering the similarity of SARS and COVID-19 on their pathogen, clinical characteristics, and therapeutic strategies, it is particularly desirable to investigate whether osteonecrosis will become a common sequela among convalescent COVID-19 patients. METHODS: This multi-strategy study was designed by integrating different research methods, such as meta-analysis, systematic review, and cross-sectional investigations to address above study objectives. At first, two meta-analyses were performed on the osteonecrosis incidence among SARS patients and the clinical data of glucocorticoid exposure among COVID-19 patients. Then, a systematic review of low-dosage glucocorticoid associated osteonecrosis and a cross-sectional investigation of glucocorticoid exposure of COVID-19 patients in Wuhan city of China were also conducted. Moreover, the pathogenesis, diagnosis, prevention, and treatment options for osteonecrosis patients with COVID-19 infection were further presented and discussed. RESULTS: Our meta-analysis showed that 32% of SARS patients had developed osteonecrosis after receiving glucocorticoid treatment with high dose, and our system review supported that low level glucocorticoid exposure might also lead to the occurrence of osteonecrosis. Similarly, 40% of COVID-19 patients had undergone glucocorticoid treatment according to our meta-analysis. The cross-sectional investigation in Wuhan city of China found that the average of cumulative glucocorticoid exposure level was 504 â€‹mg calculated by the dosage of methylprednisolone. Notably, a confirmed osteonecrosis case was identified from 1406 patients with COVID-19 during our cross-sectional investigation, implying that preventive management of osteonecrosis should be better started with regular clinical follow-up observation. CONCLUSION: Growing evidence of the glucocorticoid therapy for COVID-19 patients prompts us to establish risk-classification-based early screening and to introduce early prevention protocol of its associated osteonecrosis that will be of clinical significance in favor of improved prognosis of this disease. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: To establish risk-classification-based early screening and to introduce early prevention protocol of glucocorticoid-induced osteonecrosis will be of clinical significance in favor of improved prognosis of COVID-19.

aFGF Targeted Mediated by Novel Nanoparticles-Microbubble Complex Combined With Ultrasound-Targeted Microbubble Destruction attenuates Doxorubicin-Induced Heart Failure via Anti-Apoptosis and Promoting Cardiac Angiogenesis.

The purpose of this study was to evaluate the protective effect of acidic fibroblast growth factor targeted mediated by novel nanoparticles-cationic lipid microbubbles complex (aFGF-NP + CPMBs) combined with ultrasound targeted microbubble destruction (UTMD)on doxorubicin-induced heart failure (HF)and its mechanism. Heart failure rats induced by intraperitoneal injection with doxorubicin (DOX) to achieve cummulative dose of 15mg/kg for continuous 6 weeks showed left ventricular dysfunction, seriously oxidative stress, cardiomyocyte apoptosis, and decrease of myocardial vascular density. In contrast, aFGF-NP + CPMBs combined with UTMD therapy (3ug/kg, caudal vein injection, twice a week, 6weeks)prominently ameliorated left ventricular dysfunction by increased ejection fraction (EF) and fractional shortening (FS), decreased brain natriuretic peptide (BNP); strengthened the ability of antioxidant stress confirmed by increasing the activity of SOD and reducing the production of MDA; exerted the effect of anti-cardiomyocyte apoptosis and promotion angiogenesis by inhibited Bax expression and increased Bcl-2 expression and platelet endothelial cell adhesion molecule (CD31) expression. Taken together, the research suggested that aFGF targeted mediated by novel nanoparticles-cationic lipid microbubbles complex combined with UTMD should be a promising targeted treatment for heart failure.

Development and validation of the diagnostic scale of traditional Chinese medicine syndrome elements for diabetic kidney disease.

BACKGROUND: To construct a traditional Chinese medicine (TCM) syndrome elements diagnostic scale for diabetic kidney disease (DKD). METHODS: A total of 492 DKD patients were included in the study. TCM symptoms, signs and tongue manifestation information of the patients were collected, which constituted the items of the TCM syndrome elements diagnostic scale. Frequency dominance method was used to screen the core items. Cluster analysis and factor analysis method were used to identify the syndrome elements. Correlation coefficient and regression analysis were used to determine the syndrome elements. Regression coefficient was used to determine the scale items, and the diagnostic threshold was established by receiver operating characteristic curve. By using the above statistical methods , TCM syndrome elements diagnostic scale was constructed, and confirmed via diagnostic tests of 150 patients. RESULTS: There were 61 items of TCM diagnostic descriptions, and we kept the most useful 32 after filtering. After extracting the syndrome elements, a TCM syndrome elements diagnostic rating scale for DKD containing 9 syndrome elements was constructed, which were qi deficiency syndrome, blood deficiency syndrome, yin deficiency syndrome, yang deficiency syndrome, excessive heat syndrome, qi stagnation syndrome, damp heat syndrome, blood stasis syndrome and phlegm turbidity syndrome. A small-sample clinical validation test of the scale showed sensitivity of 78.8-100%, specificity of 84.3-100%, and accuracy of 82.7-100%. CONCLUSIONS: We constructed a TCM syndrome elements diagnostic rating scale for DKD, providing a basis for the standardized study of TCM syndromes.

Protective Effect and Mechanism of Gandou Fumu Decoction on Oxidative Damage of HepG2 Cells Induced by CuCl2 / 中国实验方剂学杂志

Objective:To explore the effect of Gandou Fumu decoction （GDFMD） on the oxidative damage of HepG2 cells induced by CuCl₂based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway. Method:CuCl₂ （200 μmol·L^-1） was used to induce a copper-loaded HepG2 cell model. HepG2 cells were divided into a blank group （HepG2 cells + blank rat serum）， a model group （HepG2 cells + CuCl₂ + normal rat serum）， a GDFMD group （HepG2 cells + CuCl₂ + GDFMD-medicated rat serum）， an inhibitor group （HepG2 cells + NVP-BEZ235 + normal rat serum）， and a GDFMD + NVP-BEZ235 group （HepG2 cells + NVP-BEZ235 + GDFMD-medicated rat serum）. ELISA method was used to determine superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） activity， and malondialdehyde （MDA） content. The expression of microtubule-associated protein 1 light chain 3 （LC3） was detected by immunofluorescence. Phospho-PI3K/PI3K （p-PI3K/PI3K）， p-Akt/Akt， p-mTOR/mTOR， Beclin-1， LC3Ⅱ/LC3Ⅰ， and p62/Actin were determined by Western blot. PI3K， Akt， mTOR， Beclin-1， LC3Ⅰ， LC3Ⅱ， p62 mRNA expression was measured by real-time polymerase chain reaction （PCR）. Result:Compared with the blank group， the model group displayed decreased activities of SOD and GSH-Px and increased content of MDA （<italic>P</italic><0.01）. Compared with the model group， the GDFMD group showed elevated activities of SOD and GSH-Px and reduced content of MDA （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the inhibitor group exhibited weakened GSH-Px activity and up-regulated content of MDA （<italic>P</italic><0.05）. Compared with the blank group， the model group showed diminished expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62， and increased expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ （<italic>P</italic><0.01）. The expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62 was elevated， and the expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ declined in the GDFMD group （<italic>P</italic><0.05，<italic> P<</italic>0.01）， while the p-PI3K/PI3K and p-mTOR/mTOR expression was down-regulated and the Beclin-1 and LC3Ⅱ/LC3 I expression was increased in the inhibitor group （<italic>P</italic><0.05， <italic>P<</italic>0.01） as compared with those in the model group. Compared with the GDFMD group， the GDFMD + NVP-BEZ235 group showed down-regulated expression of p-Akt/Akt and p-mTOR/mTOR and up-regulated expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ（<italic>P</italic><0.05， <italic>P</italic><0.01）. The expression of LC3Ⅱ protein in the model group was increased （<italic>P</italic><0.01） as compared with that in the blank group. The expression of LC3Ⅱ protein was lower in the GDFMD group than in the model group， and higher in the GDFMD + NVP-BEZ235 group than in the GDFMD group. No significant difference in the expression of PI3K， Akt， and mTOR mRNA was observed among the groups. Compared with the blank group， the model group displayed lowered expression of p62 mRNA， and elevated expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （<italic>P</italic><0.01）. Compared with the model group， the GDFMD group exhibited increased expression of p62 mRNA， and declining expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （<italic>P</italic><0.01）， while the inhibitor group showed increased expression of Beclin-1 mRNA （<italic>P</italic><0.05）. The expression of Beclin-1 and LC3Ⅱ mRNA in the GDFMD + NVP-BEZ235 group was elevated （<italic>P</italic><0.01） as compared with that in the GDFMD group. Conclusion:GDFMD may inhibit the excessive autophagy and alleviate the oxidative damage of HepG2 cells induced by CuCl₂， with the underlying mechanism related to the activation of PI3K/Akt/mTOR signalling pathway.

Mesenchymal stem cells in the treatment of spinal cord injury: Cell therapy and combination of new drugs and biomaterials / 中国组织工程研究

BACKGROUND: The treatment of spinal cord injury is mainly to improve the inflammation microenvironment of the injury site, as well as the regeneration and repair of neural function. Mesenchymal stem cells have the characteristics of easy proliferation in vitro, multi-directional differentiation, and suppression of secondary inflammation and immunomodulation, which makes the transplantation of mesenchymal stem cells for various refractory tissue injury diseases including spinal cord injury become a potential cell therapy. OBJECTIVE: To summarize the preclinical and clinical research of mesenchymal stem cells in the treatment of spinal cord injury in recent years, and put forward the problems and development directions of mesenchymal stem cells in experimental research. METHODS: We searched the articles published in Wanfang databases, CNKI and PubMed databases from 2000 to 2020. The key words were “mesenchymal stem cells, spinal cord injury, cell transplantation, immunomodulation, combination therapy, biomaterials” in Chinese and English, respectively. RESULTS AND CONCLUSION: (1) Mesenchymal stem cells from different sources play the role of anti-inflammatory, inducing axon and neuron regeneration in animal models and clinical experiments, and effectively improve the neural function of the damaged area. (2) The exosomes derived from mesenchymal stem cells show the effects of immunomodulation and angiogenesis in the disease model of spinal cord injury. (3) In order to maximize the potential of mesenchymal stem cells, exploring cell pretreatment, combined with new drugs or biological materials is the research direction of mesenchymal stem cells in the treatment of spinal cord injury.

Preparation and in vitro evaluation of fused deposition modeling 3D printed verapa-mil hydrochloride gastric floating formulations / 北京大学学报(医学版)

OBJECTIVE@#To explore the feasibility of preparing gastric floating formulations by fused de-position modeling (FDM) 3D printing technology, to evaluate the in vitro properties of the prepared FDM 3D printed gastric floating formulations, and to compare the influence of different external shapes of the formulation with their in vitro properties.@*METHODS@#Verapamil hydrochloride and polyvinyl alcohol (PVA) were used as the model drug and the excipient, respectively. The capsule-shaped and hemisphere-shaped gastric floating formulations were then prepared by FDM 3D printing. The infill percentages were 15%, the layer heights were 0.2 mm, and the roof or floor thicknesses were 0.8 mm for both the 3D printed formulations, while the number of shells was 3 and 4 for capsule-shaped and hemisphere-shaped formulation, respectively. Scanning electron microscopy (SEM) was used to observe the morpho-logy of the surface and cross section of the formulations. Gravimetric method was adopted to measure the weights of the formulations. Texture analyzer was employed to evaluate the hardness of the formulations. High performance liquid chromatography method was used to determine the drug contents of the formulations. The in vitro floating and drug release behavior of the formulations were also characterized.@*RESULTS@#SEM showed that the appearance of the FDM 3D printed gastric floating formulations were both intact and free from defects with the filling structure which was consistent with the design. The weight variations of the two formulations were relatively low, indicating a high reproducibility of the 3D printing fabrication. Above 800.0 N of hardness was obtained in two mutually perpendicular directions for the two formulations. The drug contents of the two formulations approached to 100%, showing no drug loss during the 3D printing process. The two formulations floated in vitro without any lag time, and the in vitro floating time of the capsule-shaped and hemisphere-shaped formulation were (3.97±0.41) h and (4.48±0.21) h, respectively. The in vitro release of the two formulations was significantly slower than that of the commercially available immediate-release tablets.@*CONCLUSION@#The capsule-shaped and hemisphere-shaped verapamil hydrochloride gastric floating formulations were prepared by FDM 3D printing technology successfully. Only the floating time was found to be influenced by the external shape of the 3D printed formulations in this study.

Anatomical study on reconstruction of calcaneus fibula ligament / 中国骨伤

OBJECTIVE@#To measure anatomical data of calcaneofibular ligament (CFL), relevant data of CFL attachment to provide an anatomical basis for CFL reconstruction.@*METHODS@#Twenty-seven adult ankle specimens were selected, including 11 males and 16 females, aged from 22 to 71 years old with an average of (41.6±17.2) years old;9 cases on the left side and 18 cases on the right side. The specimens reserved at least 20 cm above ankle joint and a complete foot, and exclude deformities, fractures, incomplete development and degenerative lesions. CFL was performed detailed anatomical observation, morphological parameters of CFL was measured, and coordinates of fibula side and calcaneal side of CFL in the coordinate axis were measured. The distance between fibula insertion of CFL and fibula tip, distance between calcaneal insertion of CFL and lateral calcaneal nodule, and Angle between CFL and long axis of fibula were also measured.@*RESULTS@#In these 27 specimens, CFL cases were all single bundles and the length of CFL was (32.83 ± 8.19) mm. The center point of fibula attachment in CFL was(2.87± 1.21) mm proximal with a coefficient of variation of 42.16% and (2.08±1.34) mm anteriorly with a coefficient of variation of 64.42%. The center point of calcaneal attachment region of CFL was located on coordinate axis on the distal end (15.32±5.33) mm, with a coefficient of variation of 34.79%, and the posterior part (6.38±2.15) mm, with a coefficient of variation of 33.86%. The distance between center point of fibula attachment and fibula tip was (4.81±0.82) mm. The distance between center point of calcaneal attachment area of CFL and lateral calcaneal nodules was(17.25±3.12) mm. Angle between CFL and fibula axis is (43 ±18)° .@*CONCLUSION@#According to anatomical studies, we could locate the fibula and calcaneal attachment of CFL by anatomical markers around ankle joint. However, the location of CFL attachment has a large variation, and the anatomical characteristics need to be considered in anatomical reconstruction.

Novel role of intracellular ATP in obesity pathology / 生理学报

ATP is an important energy source for cells. Traditionally, intracellular ATP levels are believed to be relatively stable and will not rise consistently in the physiological conditions. However, new studies suggest that ATP levels may rise in multiple tissues under the condition of energy surplus contributing to the metabolic disorders in obesity. However, the molecular mechanism of ATP elevation remains unknown in obesity except the increase in energy supply. Based on our experimental results and the findings reported in the literature, we discuss the cellular and molecular mechanisms by which ATP levels are regulated in cells by multiple factors, including superoxide ions, mitochondrial flash, antioxidants, anti-apoptotic molecule Bcl-xL, AMP-activated protein kinase (AMPK) and metformin. Contribution of these factors to the alteration of ATP set-point will be discussed together with their impact on insulin resistance in type 2 diabetes mellitus. With a focus on the energy surplus in obesity, we explore the mechanism of insulin resistance induced by ATP elevation and provide an answer to the contradiction between the new experimental results and the traditional viewpoint of intracellular ATP. We propose that elevation of intracellular ATP may lead to metabolic disorder in obesity through activation of a feedback mechanism that inhibits mitochondrial function.

Treatment for posterior root tear of lateral meniscus through bone tunnel suture under arthroscopy / 中国骨伤

OBJECTIVE@#To evaluate clinical effects of posterior root tear of lateral meniscus through bone tunnel suture under arthroscopy.@*METHODS@#From January 2012 to December 2014, 23 patients with posterior root tear of lateral meniscus repaired through bone tunnel suture under arthroscopy, including 15 males and 8 females, aged from 19 to 48 years old with an average age of (25.0±4.7) years old; 10 knees on the left side and 13 knees on the right side. Complications were observed, Lysholm score before and after operation at 12 months were used to evaluate clinical results, and VAS score was applied to assess pain relief. MRI was used to check recovery outcomes of lateral meniscus injury.@*RESULTS@#All patients were followed up from 13 to 24 months with an average of (17.0±4.3) months. No injury of vessels, nerve and incision infection occurred. Motion of knee joint of 19 patients reached normal, 4 patients manifested limited activity of knee joint at12 months after operation. Postoperative Lysholm score 88.52±6.48 at 12 months was higher than that of before operation 46.12±7.35; Postoperative VAS score 0.8±0.7 at 12 months was lower than that of before operation 4.3±1.6.@*CONCLUSIONS@#Bone tunnel suture under arthroscopy for the treatment of posterior root tear of lateral meniscus could relieve pain, decrease postoperative complications and obtain good clinical efficacy.

Bright carbon dots via inner filter effect for the sensitive determination of the purine metabolic disorder in human fluids.

In this paper, the high performance fluorescent carbon dots were synthesized with maleic acid, tris and benzoic acid as raw materials by one-step hydrothermal method. The obtained carbon dots with uniform size emitted strong blue fluorescence, which the maximum excitation and emission wavelengths at 250 nm and 415 nm, respectively. Under the optimum condition, it was meaningfully founded that the reaction between the carbon dots and uric acid resulting in the fluorescence quenching of the carbon dots at the emission spectrum of 415 nm. The reason was that they had a synergistic effect between the fluorescence internal filtering effect and the static quenching effect. The fluorescence internal filter effect sensing system was constructed by using uric acid as the absorbable material and carbon dots as the luminophore. Hence, a fluorescence quenching method for the determination of uric acid was established in the concentration range from 5.0 to 400 µM with the detection limit (3σ/S) of 2.26 µM. Thus, a fluorescent sensing assay for the determination of uric acid was founded and confirmed in human fluids.

Pretreatment Serum Amyloid A and C-reactive Protein Comparing with Epstein-Barr Virus DNA as Prognostic Indicators in Patients with Nasopharyngeal Carcinoma: A Prospective Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The measuring Epstein-Barr virus (EBV) DNA is an important predictor of nasopharyngeal carcinoma (NPC). This study evaluated the predictive value of pretreatment serum amyloid A (SAA) and C-reactive protein (CRP) comparing with EBV DNA in patients with NPC. MATERIALS AND METHODS: In an observational study of 419 non-metastatic NPC patients, we prospectively evaluated the prognostic effects of pretreatment SAA, CRP, and EBV DNA on survival. The primary end-point was progress-free survival (PFS). RESULTS: The median level of SAA and CRP was 4.28 mg/L and 1.88 mg/L, respectively. For the high-SAA group (> 4.28 mg/L) versus the low-SAA (≤ 4.28 mg/L) group and the high-CRP group (> 1.88 mg/L) versus the low-CRP (≤ 1.88 mg/L) group, the 5-year PFS was 64.5% versus 73.1% (p=0.013) and 65.2% versus 73.3% (p=0.064), respectively. EBV DNA detection showed a superior predictive result, the 5-year PFS in the EBV DNA ≥ 1,500 copies/mL group was obviously different than the EBV DNA < 1,500 copies/mL group (62.2% versus 77.8%, p < 0.001). Multifactorial Cox regression analysis confirmed that in the PFS, the independent prognostic factors were including EBV DNA (hazard ratio [HR], 1.788; p=0.009), tumour stage (HR, 1.903; p=0.021), and node stage (HR, 1.498; p=0.049), but the SAA and CRP were not included in the independent prognostic factors. CONCLUSION: The results of SAA and CRP had a certain relationship with the prognosis of NPC, and the prognosis of patients with high level of SAA and CRP were poor. However, the predictive ability of SAA and CRP was lower than that of EBV DNA.

Combination of Tumor Volume and Epstein-Barr Virus DNA Improved Prognostic Stratification of Stage II Nasopharyngeal Carcinoma in the Intensity Modulated Radiotherapy Era: A Large-Scale Cohort Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Little is known about combination of the circulating Epstein-Barr viral (EBV) DNA and tumor volume in prognosis of stage II nasopharyngeal carcinoma (NPC) patients in the intensity modulated radiotherapy (IMRT) era. We conducted this cohort study to evaluate the prognostic values of combining these two factors. MATERIALS AND METHODS: By Kaplan-Meier, we compare the differences of survival curves between 385 patients with different EBV DNA or tumor volume levels, or with the combination of two biomarkers mentioned above. RESULTS: Gross tumor volume of cervical lymph nodes (GTVnd, p 0 copy/mL, GTVtotal 0 copy/mL, GTVtotal ≥ 30 cm³). When patients in the low-risk group were compared with those in the high-risk group, 3-year PFS (p=0.003), LRFS (p=0.010), and DMFS (p=0.031) rates were statistically significant. CONCLUSION: Pretreatment plasma EBV DNA and tumor volume were both closely correlated with prognosis of stage II NPC patients in the IMRT era. Combination of EBV DNA and tumor volume can refine prognosis and indicate for clinical therapy.

Diagnostic value of MRI for posterior root tear of medial and lateral meniscus / 中国骨伤

<p><b>OBJECTIVE</b>To explore diagnostic value of MRI on posterior root tear of medial and lateral meniscus.</p><p><b>METHODS</b>From January 2012 to January 2016, clinical data of 43 patients with meniscal posterior root tear confirmed by arthroscopy were retrospective analyzed, including 25 males and 18 females, aged from 27 to 69 years old with an average age of(42.5±8.3)years old;27 cases on the right side and 16 cases on the left side. MRI examinations of 43 patients with tear of posterior meniscus root confirmed by knee arthroscopies were retrospectively reviewed. MRI images were double-blinded, independently, retrospectively scored by two imaging physicians. Sensitivity, specificity and accuracy of MRI diagnosis of lateral and medial meniscus posterior root tear were calculated, and knee ligament injury and meniscal dislocation were calculated.</p><p><b>RESULTS</b>Forty-three of 143 patients were diagnosed with meniscus posterior root tears by arthroscopy, including 19 patients with lateral tears and 24 patients with medial tears. The sensitivity, specificity and accuracy in diagnosis of posterior medial meniscus root tears for doctor A were 91.67%, 86.6% and 83.9% respectively, and for doctor B were 87.5%, 87.4% and 87.4%, 19 patients with medial meniscal protrusion and 2 patients with anterior cruciate ligament tear. The sensitivity, specificity and accuracy in diagnosis of posterior lateral meniscus root tears for doctor A were 73.7%, 79.9% and 79% respectively, and for doctor B were 78.9%, 82.3% and 82.5%, 4 patients with lateral meniscus herniation and 16 patients with cruciate ligament tear. Kappa statistics for posterior medial meniscus root tears and posterior lateral meniscus root tears were 0.84 and 0.72.</p><p><b>CONCLUSIONS</b>MRI could effectively demonstrate imaging features of medial and lateral meniscal root tear and its accompanying signs. It could provide the basis for preoperative diagnosis of clinicians, and be worthy to be popularized.</p>