BACKGROUND:The capability of repairing articular cartilage damage is very limited,and tissue engineering technology provides new therapeutic options for repairing damaged cartilage,in which the interaction and induction between chondrocytes,bone marrow mesenchymal stem cells,and synovial mesenchymal stem cells is the basis of autologous healing of cartilage damage. OBJECTIVE:To construct the chondrocyte-bone marrow mesenchymal stem cell-synovial mesenchymal stem cell co-culture system to simulate the in vitro microenvironment of chondrocytes,and to explore the optimal cell inoculation ratio,meanwhile to observe the effects of icariin-containing serum on the proliferation of chondrocytes and the chondrogenic differentiation of stem cells in the system. METHODS:Rat knee chondrocytes,bone marrow mesenchymal stem cells and synovial mesenchymal stem cells were extracted,cultured and identified,and a chondrocyte-bone marrow mesenchymal stem cell-synovial mesenchymal stem cell non-contact co-culture system was constructed according to different cell inoculation ratios.After 72 hours of co-culturing,the chondrocyte proliferative activity and phenotypic ability were observed,and the co-culture system with the best overall effect was selected.New Zealand white rabbits were gavaged with icariin solution(0.25 mg/mL)to prepare icariin-containing serum,and cultured in conventional complete medium(high sugar DMEM culture medium containing 10%fetal bovine serum and 1%double antibody by volume)as the control group,while the experimental group was intervened by adding 10%icariin-containing serum by volume on the basis of above.The proliferative activity of chondrocytes and the expression of collagen type II were tested for the two groups after 24 and 48 hours.The differentiation of bone marrow mesenchymal stem cells and synovial mesenchymal stem cells into chondrocytes in the co-culture system was tested by immunofluorescence staining after 14 days. RESULTS AND CONCLUSION:(1)The three kinds of cells grew normally adherently to the wall in different ratios of co-culture,where chondrocytes showed the best proliferative activity and phenotypic ability in the co-culture system when chondrocytes:bone marrow mesenchymal stem cells:synovial mesenchymal stem cells=2:1:1.(2)Compared with the control group,the proliferative activity and type II collagen expression of chondrocytes in the experimental group were significantly increased after 24 hours(P<0.01),and the two groups still had difference after 48 hours(P<0.05).The two groups showed obvious chondrogenic differentiation of bone marrow mesenchymal stem cells and synovial mesenchymal stem cells after 14 days(P<0.01),and some of the cells appeared round or oval,and the cytoplasmic type II collagen immunofluorescence staining was positive.The fluorescence intensity of the experimental group was significantly higher than that of the control group(P<0.01).(3)The results showed that the chondrocyte-bone marrow mesenchymal stem cell-synovial mesenchymal stem cell co-culture system could be successfully established by the non-contact co-culture method,and the best chondrocyte proliferative activity and phenotypic ability could be obtained when the cell ratio was 2:1:1.Icariin-containing serum had the promoting effect on chondrocyte proliferation,and chondrogenic differentiation of bone marrow mesenchymal stem cells and synovial mesenchymal stem cells in the system.

Renal interstitial fibrosis（RIF） is the main pathological manifestation of chronic kidney disease. Due to the complexity of the mechanism， there is no specific treatment for RIF in clinical practice. The abnormal activation of the phosphatidylinositol 3-kinase （PI3K）/protein kinase B（Akt） signaling pathway and the activation of downstream target genes are key drivers of RIF induction and progression. Traditional Chinese medicine has the characteristics of precise efficacy and minimal toxic side effects， and the occurrence and development of RIF can be regulated by multiple targets and mutual coordination. This review focuses on the PI3K/Akt signaling pathway and summarizes the potential targets and regulatory mechanisms of traditional Chinese medicine in the treatment of RIF. It is found that various effective ingredients （such as sinomenine， mangiferin， coumarin derivates from Hydrangea paniculata， etc.） and formulas （such as Fushengong decoction， Qi-Bang-Yi-Shen formula， etc.） of traditional Chinese medicine can inhibit fibroblast proliferation， improve inflammation and oxidative stress， maintain mitochondrial stability， and slow down ferroptosis through this pathway， thereby delaying the occurrence and progression of RIF.

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.

BACKGROUND:Kidney deficiency and blood stasis syndrome are common traditional Chinese medicine syndromes observed in knee osteoarthritis,which serve as fundamental pathogenesis factors.There exists a significant connection between the two.Previous studies have demonstrated that kidney deficiency and blood stasis syndrome effectively contribute to knee joint cartilage degeneration and the progression of knee osteoarthritis.However,the mechanisms underlying the promotion of knee joint cartilage damage remain unclear and require further investigation. OBJECTIVE:To investigate the influence of kidney deficiency and blood stasis syndrome on the progression of knee osteoarthritis in Sprague-Dawley rats. METHODS:Sixteen Sprague-Dawley rats were randomly divided into two groups:a model observation group and a control group,with eight rats in each group.Animal models of kidney deficiency were induced by ovary removal in the model observation group,while the control group was given a sham procedure for ovarian removal.Two months after modeling,both groups underwent modified HULTH surgery to induce knee osteoarthritis.One week after modified HULTH surgery,the model observation group was subcutaneously given adrenaline hydrochloride to make blood stasis models,while the control group was subcutaneously given normal saline.At the 5th week after modified HULTH surgery,blood rheology,coagulation parameters,triiodothyronine,tetraiodothyronine,and estradiol levels were measured.Knee joint X-ray images were taken,and knee joint sections were stained with safranin O-fast green,hematoxylin-eosin,and immunohistochemistry. RESULTS AND CONCLUSION:Compared with the control group,the model observation group exhibited significant increases in whole blood viscosity at low,medium,and high shear rates,as well as increased plasma viscosity.Fibronectin levels in the coagulation parameters were significantly increased,while prothrombin time and activated partial thromboplastin time were significantly decreased.Triiodothyronine,tetraiodothyronine,and estradiol levels were all significantly decreased.Radiographic results showed that the model observation group exhibited more severe degree of knee joint space narrowing and surface roughness,with the appearance of high-density shadows.Hematoxylin-eosin and safranin O-fast green staining demonstrated more severe cartilage damage in the model observation group,with significantly higher OARSI and Mankin scores compared with the control group.Compared with the control group,immunohistochemistry results showed a significant reduction in the expression of extracellular matrix type II collagen and aggrecan protein in the cartilage of the model observation group rats.Moreover,there was a significant increase in the expression of matrix metalloproteinase 13 and aggrecanase 5,which are inflammatory factors.These results indicate that the Sprague-Dawley rat model of knee osteoarthritis with kidney deficiency and blood stasis was successfully established.Kidney deficiency and blood stasis syndrome further aggravate cartilage extracellular matrix degradation and cartilage degeneration by promoting the expression of inflammatory factors,thereby promoting the progression of knee osteoarthritis in rats.

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 .

To analyze the correlation between lipid profile and disease activity in patients with inflammatory bowel disease (IBD).A total of 307 Crohn′s disease (CD) patients, 232 ulcerative colitis (UC) patients and 165 healthy subjects from the same geographic region were included. Total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and lipoprotein(a)[Lp(a)] were retrieved from their medical records. Crohn disease activity index (CDAI) and Mayo scores were calculated as measurement of disease severity for CD and UC separately. Patients with CD and UC had lower TC, TG, HDL-C and LDL-C levels than those in control group ( P<0.05). Additionally, CDAI was negatively associated with TC, HDL-C and LDL-C levels ( r=-0.218, -0.210, -0.176, P<0.05), while TG level was not associated with CDAI. Mayo scores was not significantly associated with TC, HDL-C, LDL-C and TG. Patients with CD had higher Lp(a) levels than those in UC and control group ( P<0.05). Furthermore, patients with active CD had higher Lp (a) levels than those with inactive disease ( P<0.05).The Lp(a) levels in CD patients were positively associated with CDAI ( r=0.151, P<0.05), while Lp(a) level in UC group was nor assocriated with Mayo score. Patients with IBD have dyslipidemia and lipid profile is associated with disease activity in CD patients.

Objective:Mesenteric fat hypertrophy is present in about a quarter of Crohn′s disease (CD) patients and it can be easily detected by bowel ultrasound (US). The purpose of this research was to assess the correlation between mesenteric fat hypertrophy and behavior and activity of CD.Methods:A total of 89 CD patients who admitted to the First Affiliated Hospital of Nanjing Medical University from August 2018 to November 2019 were recruited in this study. The total CD patients were divided into two groups depending on with or without mesenteric fat hypertrophy by US tests. Crohn′s disease activity index (CDAI), simplified endoscopic score for Crohn′s disease (SES-CD), serum inflammatory indicators and fecal calprotectin were assessed.Results:Mesenteric fat hypertrophy was significantly associated with stricturing behavior (B2, P<0.01). CDAI ( P=0.002) , blood platelet ( P=0.001) , C-reactive protein ( P=0.024) , fecal calprotectin ( P=0.004) and bowel wall thickness ( P<0.01) in patients with mesenteric fat hypertrophy were significantly higher than those without, but not the erythrocyte sedimentation rate ( P=0.110) and SES-CD ( P=0.115) . Serum albumin ( P=0.001) in patients with mesenteric fat hypertrophy was lower than that in patients without mesenteric fat hypertrophy. Conclusion:Mesenteric fat hypertrophy is correlated with intestinal stenosis and disease activity in patients with Crohn′s disease.

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.