BACKGROUND:Previous studies of the research group have found that icariin-containing serum can delay the progression of knee osteoarthritis,promote chondrocyte proliferation and stem cell cartilage differentiation in rats,but there is still a lack of sufficient basis for clinical application.OBJECTIVE:To investigate the repair effect of icariin-containing serum on lipopolysaccharide-induced inflammatory damage of human chondrocytes.METHODS:The effect of icariin-containing serum on chondrocyte viability was detected by cell counting kit-8 method,and the optimal volume fraction and time of icariin-containing serum were screened.The cells were then divided into blank group,lipopolysaccharide group,icariin-containing serum group and lipopolysaccharide+icariin-containing serum group.After grouping,immunofluorescence was used to detect the secretion of type Ⅱ collagen in chondrocytes in each group.Real-time PCR was used to detect the expression of related genes.RESULTS AND CONCLUSION:(1)Icariin-containing serum showed good biosafety characteristics for human chondrocytes,and the cell viability reached the highest level after 48 hours of intervention when the icariin-containing serum volume fraction was 15％,and the follow-up experiments were carried out according to the conditions.(2)Compared with the blank group,lipopolysaccharide significantly inhibited the expression of type Ⅱ collagen in chondrocytes,while icariin-containing serum showed a positive mobilization effect,which could effectively promote the secretion of type Ⅱ collagen in chondrocytes under normal and inflammatory conditions.(3)The mRNA expression of type Ⅱ collagen and Aggrecan in chondrocytes decreased significantly under lipopolysaccharide stimulation,and the mRNA expression levels of matrix metalloproteinase 13 and a disintegrin-like and metalloproteinase with thrombospondin motifs-5 increased significantly.Icariin-containing serum promoted the mRNA expression of type Ⅱ collagen in human chondrocytes under inflammatory conditions and reduced the mRNA expression of matrix metalloproteinase 13 and a disintegrin-like and metalloproteinase with thrombospondin motifs-5 in chondrocytes after lipopolysaccharide intervention.Therefore,these findings indicate that the icariin-containing serum has good safety for human chondrocytes and plays an important role in maintaining the normal physiological functions of chondrocytes,promoting the synthesis of extracellular matrix,and inhibiting the secretion of inflammatory factors.

BACKGROUND:Previous studies of the research group have found that icariin-containing serum can delay the progression of knee osteoarthritis,promote chondrocyte proliferation and stem cell cartilage differentiation in rats,but there is still a lack of sufficient basis for clinical application.OBJECTIVE:To investigate the repair effect of icariin-containing serum on lipopolysaccharide-induced inflammatory damage of human chondrocytes.METHODS:The effect of icariin-containing serum on chondrocyte viability was detected by cell counting kit-8 method,and the optimal volume fraction and time of icariin-containing serum were screened.The cells were then divided into blank group,lipopolysaccharide group,icariin-containing serum group and lipopolysaccharide+icariin-containing serum group.After grouping,immunofluorescence was used to detect the secretion of type Ⅱ collagen in chondrocytes in each group.Real-time PCR was used to detect the expression of related genes.RESULTS AND CONCLUSION:(1)Icariin-containing serum showed good biosafety characteristics for human chondrocytes,and the cell viability reached the highest level after 48 hours of intervention when the icariin-containing serum volume fraction was 15％,and the follow-up experiments were carried out according to the conditions.(2)Compared with the blank group,lipopolysaccharide significantly inhibited the expression of type Ⅱ collagen in chondrocytes,while icariin-containing serum showed a positive mobilization effect,which could effectively promote the secretion of type Ⅱ collagen in chondrocytes under normal and inflammatory conditions.(3)The mRNA expression of type Ⅱ collagen and Aggrecan in chondrocytes decreased significantly under lipopolysaccharide stimulation,and the mRNA expression levels of matrix metalloproteinase 13 and a disintegrin-like and metalloproteinase with thrombospondin motifs-5 increased significantly.Icariin-containing serum promoted the mRNA expression of type Ⅱ collagen in human chondrocytes under inflammatory conditions and reduced the mRNA expression of matrix metalloproteinase 13 and a disintegrin-like and metalloproteinase with thrombospondin motifs-5 in chondrocytes after lipopolysaccharide intervention.Therefore,these findings indicate that the icariin-containing serum has good safety for human chondrocytes and plays an important role in maintaining the normal physiological functions of chondrocytes,promoting the synthesis of extracellular matrix,and inhibiting the secretion of inflammatory factors.

ObjectiveTo evaluate the safety/efficacy of compound Kushen injection （CKI） by zebrafish model and explore the possible mechanism. MethodsZebrafish were exposed to different concentrations of CKI solution， and the mortality rate after 24 h was calculated. After exposure to sublethal concentration （10） and safe dose concentration （0） for 24 h， the safety of CKI was evaluated based on acridine orange staining for the liver， liver area changes， and liver histological sections. The inflammatory bowel disease （IBD） model of zebrafish was established with 2，4，6-trinitrobenzenesulfonic acid sol （TNBS）. The efficacy of CKI in the treatment of IBD was evaluated by the changes in the area of neutral red staining， the number of neutrophils， the area of alcian blue staining， and the contents of tight junction protein （Occludin）， zonula occludens-1 （ZO-1）， tumor necrosis factor（TNF）-α， and prostaglandin E₂（PGE₂） in the intestine of zebrafish. The mechanism of CKI in the treatment of IBD was predicted by network pharmacology and verified by real-time polymerase chain reaction（Real-time PCR）. ResultsIn the safety evaluation， compared with the blank group， the sublethal concentration of CKI could cause liver cell apoptosis， liver area reduction， loose and disordered arrangement of liver cell structure， obvious vacuolation， and other pathological characteristics of zebrafish， while these indicators showed no significant changes under safe dose conditions. In the effectiveness evaluation， compared with the model group， the safe dose of CKI could increase the neutral red staining area of the intestine in a dose-dependent manner and improve the intestinal phagocytosis function. It could reduce the accumulation of intestinal neutrophils， increase the alcian blue staining area， enhance intestinal goblet cell secretion， improve the contents of Occludin and ZO-1， and decrease the contents of TNF-α and PGE₂. Network pharmacology analysis identified 288 potential targets for CKI treatment of IBD. The top five targets obtained by protein-protein interaction （PPI） network analysis were epidermal growth factor receptor A（EGFRA）， protein kinase B1（Akt1）， heat shock protein 90（HSP90AA1）， homologous oncogene of avian sarcoma virus（SRC）， and protooncogene （JUN）. Kyoto encyclopedia of genes and genomes（KEGG） results showed that CKI may be related to the Wnt signaling pathway and cholinergic synaptic pathway in the treatment of IBD， and Real-time PCR results verified the above pathways. ConclusionExcessive use of CKI can induce obvious liver injury in zebrafish， while the rational use of CKI does not cause obvious toxic reactions and can achieve a therapeutic effect on IBD by regulating the Wnt pathway.

ObjectiveBased on the nuclear factor erythroid 2 related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway， this paper explores the effect of Sinisan （SNS） on liver oxidative stress injury in cholestatic hepatitis rats and its mechanism. MethodThirty 6-week-old male SD rats were randomly divided into a control group， model group， low and high dose groups of SNS （2.5 and 5 g·kg^-1） and ursodeoxycholic acid group （UDCA， 63 mg·kg^-1）， with six rats in each group. Rats were administrated for seven consecutive days. On the 5th day， the control group was given olive oil of 10 mL·kg^-1， and the other groups were given alpha-naphthalene isothiocyanate （ANIT） of 80 mg·kg^-1. The serum biochemical indicator levels of cholestasis and the content of antioxidant factors in rat liver were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in liver tissue. The relative mRNA and protein expressions of Nrf2， HO-1， and quinone oxidoreductase 1 （NQO1） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the control group， the model group showed a significant increase in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）. There were obvious pathological changes in the model group such as the disordered arrangement of hepatocytes， obvious congestion and necrosis in the portal area， infiltration of inflammatory cells， and destruction of the interlobular bile duct. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly down-regulated in the model group （P<0.05， P<0.01）. Compared with the model group， the groups of SNS showed a significant decrease in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）， and the pathological liver injury was obviously improved. The necrotic area was reduced， and the infiltration of inflammatory cells was decreased. In addition， there was a small amount of extravasated blood in the interlobular vein. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly up-regulated （P<0.05， P<0.01）. ConclusionSNS can significantly improve liver injury in cholestatic hepatitis rats， and its mechanism may be related to the inhibition of oxidative stress response mediated by the Nrf2/HO-1 signaling pathway.

Objective To analyze the correlation between morphological and dosimetric parameters in patients with esophageal cancer at different locations using multi-to-multi double screening stepwise regression method,and to make simple predictions.Methods A retrospective analysis was conducted on 105 patients with advanced esophageal cancer who underwent radiotherapy at the First Affiliated Hospital of Fujian Medical University from 2019 to 2021.Morphological parameters of organs-at-risk were collected from CT images,and intensity-modulated radiotherapy plans were developed using Raystation4.7.The prescription doses for PTV-G and PTV-C were 60 Gy/30 F and 54 Gy/30 F,respectively.Multi-to-multi double screening stepwise regression method was employed to analyze the correlation between morphological and dosimetric parameters in esophageal cancer patients,and some preliminary predictions were provided.Results The dosimetric volume parameters of the lungs and heart were correlated with PTV-G volume,PTV-G length,PTV-G cross-sectional area,left and right lung volumes,lung length and total lung volume(P<0.05).For upper thoracic esophageal cancer,dosimetric volume parameters of the lungs and heart were correlated with PTV-G volume,PTV-G length,and right lung volume(P<0.05).For middle thoracic esophageal cancer,dosimetric volume parameters of the lungs,heart,and spinal cord were correlated with PTV-G volume,PTV-G length,PTV-G cross-sectional area,left and right lung volumes,and lung length(P<0.05).For lower thoracic esophageal cancer,dosimetric volume parameters of the lungs,heart,and spinal cord were correlated with PTV-G volume,PTV-G length,right lung volume,and lung length(P<0.05).Conclusion For patients with tumors at different locations,both overall and segmental analyses should be considered to balance therapeutic effect and side effects of radiotherapy,thereby maximizing the benefits for tumor patients.

Objective To analyze the correlation between morphological and dosimetric parameters in patients with esophageal cancer at different locations using multi-to-multi double screening stepwise regression method,and to make simple predictions.Methods A retrospective analysis was conducted on 105 patients with advanced esophageal cancer who underwent radiotherapy at the First Affiliated Hospital of Fujian Medical University from 2019 to 2021.Morphological parameters of organs-at-risk were collected from CT images,and intensity-modulated radiotherapy plans were developed using Raystation4.7.The prescription doses for PTV-G and PTV-C were 60 Gy/30 F and 54 Gy/30 F,respectively.Multi-to-multi double screening stepwise regression method was employed to analyze the correlation between morphological and dosimetric parameters in esophageal cancer patients,and some preliminary predictions were provided.Results The dosimetric volume parameters of the lungs and heart were correlated with PTV-G volume,PTV-G length,PTV-G cross-sectional area,left and right lung volumes,lung length and total lung volume(P<0.05).For upper thoracic esophageal cancer,dosimetric volume parameters of the lungs and heart were correlated with PTV-G volume,PTV-G length,and right lung volume(P<0.05).For middle thoracic esophageal cancer,dosimetric volume parameters of the lungs,heart,and spinal cord were correlated with PTV-G volume,PTV-G length,PTV-G cross-sectional area,left and right lung volumes,and lung length(P<0.05).For lower thoracic esophageal cancer,dosimetric volume parameters of the lungs,heart,and spinal cord were correlated with PTV-G volume,PTV-G length,right lung volume,and lung length(P<0.05).Conclusion For patients with tumors at different locations,both overall and segmental analyses should be considered to balance therapeutic effect and side effects of radiotherapy,thereby maximizing the benefits for tumor patients.

ObjectiveTo study the mechanism of matrine in the treatment of inflammatory bowel disease （IBD） based on the zebrafish model and network pharmacology. MethodThe IBD model of zebrafish was established using 2，4，6-trinitro-benzenesulfonicacid （TNBS）， and the intestinal phagocytic function， goblet cell secretion， and neutrophil aggregation were evaluated using neutral red staining， alcian blue staining， and neutrophil number changes. Changes in tumor necrosis factor （TNF）-α and cholecystokinin （CCK） content in zebrafish were determined by using relevant reagent kits. Network pharmacology and molecular docking techniques were used to predict the potential mechanism of matrine in the treatment of IBD. Gene expression of relevant targets was verified through Real-time polymerase chain reaction （Real-time PCR）. ResultCompared with the model group， the matrine administration group can increase the neutral red staining area in a dose-dependent manner and improve intestinal phagocytic function（P<0.05，P<0.01）. It can reduce the staining area of alcian blue and affect the secretion of intestinal goblet cells（P<0.01）. It can reduce the number of neutrophil granulocytes， relieve its aggregation， significantly reduce TNF-α content（P<0.01）， and increase the CCK content. Network pharmacology analysis identifies 28 potential targets for matrine in the treatment of IBD. The top five targets by protein-protein interaction （PPI） network analysis are CHRNA7， DRD1， CHRNA4， SLC6A3， and GRM5. The Kyoto encyclopedia of genes and genomes （KEGG） results show that the treatment of IBD with matrine may be related to neuroactive ligand-receptor interaction， cholinergic synapse， and neutrophil extracellular trap formation. Real-time PCR results show that matrine can affect the expression level of related target genes. Conclusionmatrine has a certain therapeutic effect on IBD and can affect the inflammatory response of IBD. Its therapeutic effect may be related to neuroactive ligand-receptor interaction and other pathways.

OBJECTIVE To separate and identify the chemical constituents of the n-butanol fraction from the stems of Clerodendrum trichotomum ，and to investigate their antitumor activities in vitro . METHODS The ethanol extracts were obtained with 85% ethanol from dried stems of C. trichotomum. After dispersed with water ，ethanol extracts were distributed by petroleum ether，ethyl acetate and n-butanol in turn ，then concentrated under reduced pressure to obtain the fractions of each extraction part . The n-butanol fraction from the stems of C. trichotomum was isolated and purified by macroporous resin D 101 column chromatography and various chromatographic techniques including silica gel ，hydroxypropyl glucan gel and Toyopearl HW -40F macroporous resin and so on . The structures of them were identified by physical and chemical properties ，MS and NMR . All these compounds were evaluated for cytotoxic activities against 4 kinds of human tumor cells such as cultured K 562，MCF-7，A549 and HepG2，using the MTT assay . RESULTS Fourteen chemical constituents were isolated and identified as teuvincenone B （1）， uncinatone（2），villosin C （3），syringaresinol（4），syringaresinol-4ʹ-O-β-glucopyranoside（5），3，12-O-β-D-diglucopyranosyl-11，16- dihydroxyabieta-8，11，13-triene（6），glypentoside C （7），martynoside（8），isomartynoside（9），2-（4-hydroxyphenyl）ethanol-O-β-D- glucopyranosyl-（1→2）-O- β -D-glucopyranoside（10），3，4-dimethoxyphenyl-1-O- β -D-apiofuranosyl （1→2） - β -D- glucopyranoside（11）， 2，6-dimethoxy-4-hydroxy-1-O- β -D- glucopyranoside（12），adenosine（13）and cistanoside F （14）. In vitro anti-tumor activity studies showed that compounds 1-3 showed certa in inhibitory activities against tumor cellproliferation，among which compound 2 displayed the strongest inhibitory activity against MCF -7，A549 and HepG 2 cells，and their IC 50 values were 25.00，22.34 and 12.50 μmol/L respectively ；only compound 3 showed stronger inhibitory activity against K562 cell with IC 50 of 28.41 μmol/L. CONCLUSIONS Among them ，compounds 10 to 13 are isolated from genus Clerodendrum for the first time ，compounds 4，5，14 were isolated from C. trichotomum for the first time . The abietane diterpenoids （compounds 1-3）have better inhibitory activities against above four tumor cell lines .

Objective:To explore the effectof tumor volume on pulmonary dose-volume parameters by intensity-modulated radiation therapy (IMRT) in non-small cell lung cancer (NSCLC),and to provide a basis for pulmonary dose parameters in IMRT treatment.Methods:A total of 204 patients with NSCLC received IMRT were retrospectively analyzed from June,2009 to October,2013.The prescribed dose of planning target volume (PTV) for primary tumor was 60-66Gy (2.00-2.25 Gy,27-33 times in all).The fractional volume percent of the lung received a dose ＞5 or 20 Gy (V5,V20),and absolute volume of lung received a dose ＜5 Gy (AVS5).The mean lung dose (MLD) in normal tissues were analyzed.Regression model curve was used to analyze them along with the change of primary tumor volume.Results:With the increase in lung tumor volume,the V5,V20 and MLD presented quadratic equation curve,and AVS5 presented logarithmic equation.When the tumor volume,less than a certain value (294.6,283.2,304.9 cm3,respectively),the V5,V20 and MLD increased with tumor size and presented an increased quadratic curve;when the tumor volume was higher than a certain value (294.6,283.2,304.9 cm3 respectively),the V5,V20 and MLD was declined.The AVS5 was declined in a logarithmic curve along with the increase of tumor volume.Conclusion:With the increase in lung tumor volume,the change in rule ofV5,V20,MLD and AVS5 is not completely equivalent.When the tumor volume exceeds a certain boundary value (about 300 cubic centimeter),the corresponding tumor diameter is about 7-8 cm.In addition to the focus on pulmonary V5,V20 and MLD,we should also pay more attention to AVS5 restrictions in establishment ofIMRT in NSCLC.

Objective: To calculate out the Hausdorff distance based on the scripting in RayStation treatment planning system, which was then applied in measuring the deformation error of brain stem during image automatic registration between CT and MR. Methods: Scripting was edited in RayStation system (version 4.7) by using IronPython. The set of point coordinates on the contour of any two region of interest (ROI) had been found firstly, then the Hausdorff distance between the two point sets was calculated out. A graphical user interface (GUI) was designed by using XAML to acquire the visualized output of Hausdorff distance. GUI appeared when the script was run, where two ROIs was selected, then the corresponding Hausdorff distance and the running time were displayed by pressing the "Calculate" button. Results: The mean Hausdorff distance of brain stem in 20 patients with head and neck neoplasms was 1.20 cm while the mean elapsed time was 11.01s. Conclusions Hausdorff distance of any two ROIs can be calculated out by using the developed method. GUI is designed to realize the visual interaction with RayStation system. Therefore, the RayStation system satisfies the demands of Hausdorff distance calculation in both clinical and research work.