Design and implementation of ultrasound therapy device for rabbit urethral scars / 国际生物医学工程杂志

Objective:To design a specialized ultrasound therapeutic device for rabbit urethral scars and to verify its applicability and effectiveness.Methods:New Zealand male rabbits were used as the experimental objects, and the ultrasound therapeutic instrument was customized according to the structure and size of the rabbit penises. The ultrasound therapeutic instrument included the ultrasound pulse emission and control system, the final-stage amplifier, and the ultrasound probe. Firstly, the ultrasound probe was designed according to the size and structure of rabbit penises, and the parameters of the ultrasound probe were determined by COMSOL finite element simulation and actual testing of the sound field distribution. Secondly, the driving circuit of the ultrasound probe was designed according to the parameters of the elements. Then the ultrasound pulse emission and control system based on the field-programmable gate array (FPGA) and the serial screen were designed. Subsequently, the ultrasound therapeutic instrument was subjected to a performance test and a safety test. The ultrasound therapeutic instrument was constructed to include the ultrasound amplifier and the ultrasound probe. Finally, a rabbit urethra reconstruction model was constructed, and eight white rabbits were randomly divided into a model group and an experimental group. The rabbits in the experimental group received the ultrasound therapeutic instrument for treatment of the urethra immediately, with an ultrasound frequency of 2 MHz, a pulse interval of 10 ms, and an output sound intensity of 0.73 W/cm 2. The treatment was performed twice a week (on Tuesday and Thursday), with 10 min of irradiation each time, lasting for four weeks. The rabbits in the model group did not receive any treatment. The area percentage of transforming growth factor-β1 (TGF-β1), matrix metalloproteinase-2 (MMP-2), and tumor necrosis factor-α (TNF-α) staining-positive areas in rabbit urethral tissues were quantitatively analyzed, and the urethral circumference was calculated using Image J software. Results:Due to the addition of sound-absorbing materials, the sound pressure distribution in the treatment chamber was more uniform, and the average value of the standing wave ratio was 1.11, indicating that the structural design met the design requirements. In the overall performance test, the natural focal position of the three ultrasonic transducers was 10 mm, and the consistency of the sound field distribution meet the experimental requirements. The relationship between the peak sound pressure of each transducer and the power supply voltage was close to linear. The output sound intensity ranged from 0.35 to 0.74 W/cm 2, which met the experimental requirements. With the ultrasound output, the temperature of the test point increased slowly, and this experiment could increase the temperature of the tissue by up to 3.3 ℃, which would not lead to thermal damage to the tissue. Animal experiment results showed that the immunopositive area fraction of TGF-β1 in the urethral tissues of rabbits in the experimental group [(4.21 ± 1.32)%] was smaller than that of the model group [(8.53 ± 3.43)%] ( t = ?4.24, P < 0.001). The immunopositive area fraction of TNF-α in the urethral tissues of rabbits in the experimental group [(5.14 ± 2.72)%] was smaller than that of the model group [(7.23 ± 1.57)%] ( t = ?3.37, P < 0.05). The MMP-2 level in the urethral tissue of rabbits in the experimental group [(10.65 ± 2.24)%] was higher than that of the model group[(6.98 ± 2.74)%] ( t = 2.19, P < 0.05). The urethral circumference [(12 209 ± 2 743) μm] was higher than that of the model group [(10 127 ± 2 237) μm] ( t = 15.46, P < 0.05). Conclusions:An ultrasound therapeutic instrument dedicated to rabbit urethral scars has been successfully designed and can be used for the study of ultrasound treatment of rabbit urethral scars.

Therapeutic Efficacy of Jingangteng Capsules Combined with Guizhi Fuling Capsules in the Treatment of Chronic Pelvic Inflammation and Their Effect on Serum Granulocyte-Macrophage Colony-Stimulating Factor and Matrix Metalloproteinase 2 Levels / 广州中医药大学学报

Objective To investigate the efficacy of Jingangteng Capsules combined with Guizhi Fuling Capsules(GFC)for the treatment of patients with chronic pelvic inflammation of damp-heat and stasis obstruction type and to observe their effects on serum granulocyte-macrophage colony-stimulating factor(GM-CSF)and matrix metalloproteinase 2(MMP-2)levels.Methods Ninety patients with chronic pelvic inflammation of damp-heat and stasis obstruction type were randomly divided into the combined group and the GFC group,with 45 patients in each group.Patients in the GFC group were treated with Guizhi Fuling Capsules,while patients in the combined group were given Jingangteng Capsules together with GFC.The treatment period lasted for 2 weeks and then one-month follow-up was conducted.The changes of traditional Chinese medicine(TCM)scores,serum GM-CSF and MMP-2 levels in the two groups were observed before and after treatment.And the clinical efficacy,time for the relief of symptoms,recurrence of disease and occurrence of adverse reactions in the two groups were compared.Results(1)After 2 weeks of treatment,the total effective rate of the combined group was 93.33%(42/45),and that of the GFC group was 66.67%(30/45).The intergroup comparison showed that the therapeutic effect of the combined group was significantly superior to that of the GFC group when comparing the two groups(P＜0.01).(2)After treatment,the scores of TCM symptoms of lower abdominal pain,lumbosacral pain,leukorrhagia,profuse menstruation,dysmenorrhea,and fatigue in both groups were significantly lower than those before treatment(P＜0.05),and the reduction of TCM syndrome scores in the combined group was significantly superior to that in the GFC group(P＜0.05).(3)The time for leucorrhea recovering normal and the time for the relief of lower abdominal distension and abdominal pain in the combined group were significantly shorter than those in the GFC group after treatment(P＜0.01).(4)After treatment,the serum serological indicators of GM-CSF and MMP-2 levels in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the reduction of serum GM-CSF and MMP-2 levels in the combined group was significantly superior to that in the GFC group(P＜0.05 or P＜0.01).(5)The recurrence rate and the incidence rate of adverse reactions in the combined group were 11.11%(5/45)and 13.33%(6/45),respectively,and were significantly lower than those in the GFC group[all being 35.56%(16/45)],the differences being all statistically significant(P＜0.05 or P＜0.01).Conclusion Jingangteng Capsules combined with Guizhi Fuling Capsules can significantly enhance the clinical efficacy of the patients with chronic pelvic inflammatory of damp-heat and stasis obstruction type,effectively shorten the time for the relief of symptoms,and decrease the serum GM-CSF and MMP-2 levels.

Effects of salidroside on proliferation and migration of fibroblastoid synovial cells in rheumatoid arthritis by regulating miR⁃20a⁃5p/TIMP2 axis / 安徽医科大学学报

Objective @#To investigate ellect of salidroside on the function and activation of rheumatoid arthritis fibroblast-like synoviocyte(HFLS-RA)by regulating the miR-20a-5p/tissue inhibitor of metalloproteinase-2(TIMP2) axis.@*Methods@#HFlS-RA cells were used as the research object. HFlS-RA cells were separated intocontrol group, tumor necrosis factor-a (TNF-a) group, salidroside group, inhibitor NC group, miR-20a-5p inhibitor group, salidroside + mimic NC group, and salidroside + miR-20a-5p mimic group. qRT-PCR was applied to deteet the expression of miR-20a-5p in HFIS-RA cells ; enzyme-linked immunosorbent assay( ELISA) was applied todetect the levels of interleukin-18 ( lL-1β) and IL-6 in the supermatant of HFLS-RA cells: cell counting kit-8(CCK-8) method and 5-ethynyl-2 '-deoxyuridine ( EdU) staining were applied to detect HFLS-RA cell proliferation ; scratch experiment was applied to detect HilS-RA cell migration; Western blot was applied to detect the ex.pression of 'TlMP2, CyclinD1, and matrix metalloproteinase ( MMP ) -9 proteins in HFLS-RA cells; double lucifer.ase was applied to verify the relationship between miR-20a-5p and TIMP2. @*Results@#Compared with the control group, the expression of miR-20a-5p, the levels of lL-1β and IL-6, 0Dso value, EdU positive cell rate, scratchhealing rate, and the expression of CyclinDl and MMP-9 proteins in the TNF-α group increased, the expression of TlMP2 protein decreased ( P <0. 05 ) ; compared with the TNF-α group, the expression of miR-20a-5p, the levelsof lL.-1β and IL-6, OD450 value, EdU positive cell rate, scratch healing rate, and CyclinD1 and MMP-9 proteinsexpression decreased, the expression of TlMP2 protein increased in salidroside group ( P <0. 05 ); compared withthe 'T'NF -a group and inhibitor NC group, the expression of miR-20a-5p, the levels of IL-1 β and IL.-6, OD450 val-ue, EdU positive cell rate, seratch healing rate, and the expression of CyclinDl and MMP-9 proteins in the miR.20a-5p inhibitor group decreased, the expression of TlMP2 protein increased ( P <0. 05 ); compared with the sali.droside group and the salidroside + mimic NC group, the expression of miR-20a-5p, the levels of IL-1 β and IL-6 ,OD.so value, EdU positive cell rate, scratch healing rate, and the expression of CyelinD1 and MMP-9 proteins inthe salidroside + miR-20a-5p mimic group increased, the expression of TIMP2 protein decreased ( P < 0. 05 )There was a targeted regulatory relationship between miR-20a-5p and TIMP2. @*Conclusion@#Salidroside may inhibit TNF-α-induced HFS-RA cell proliferation , migration and infammatory response by regulating miR-20a-5p/TIMP2.

Effect of vascular endothelial growth factor-A165 on scleral remodeling of guinea pigs with form-deprivation myopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of the intravitreal injection of vascular endothelial growth factor-A165(VEGF-A165)on the scleral remodeling of guinea pigs with form-deprivation myopia(FDM).METHODS: A total of 120 tricolor guinea pigs, aged three weeks, were randomly divided into 6 groups, with 20 in each group. The blank group did not undergo any intervention. In the FDM group, only the FDM model was established. In the phosphate buffer saline(PBS)group, 2.5 μL of PBS was injected into the vitreous cavity before establishing the FDM model. In the 1ng group, 5ng group, and 10ng group, VEGF-A165 was injected into the vitreous cavity at concentrations of 1, 5 and 10ng, respectively, before the establishment of the FDM model. The FDM model was established by covering the right eyes of guinea pigs with translucent balloons for 14d. The diopter and axial length of the right eyes were measured before and after covering. After 14d, the content of dopamine(DA)in retina was measured by high performance liquid chromatography. Additionally, the mRNA and protein expression levels of matrix metalloproteinase-2(MMP-2), tissue inhibitor of matrix metalloproteinase-2(TIMP-2), transforming growth factor(TGF)-β1, TGF-β2 and α-smooth muscle actin(α-SMA)in sclera were detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blot.RESULTS: Before covering, there were no significant differences in the diopter and axial length of the right eyes of guinea pigs in all groups(P&#x003E;0.05). After 14d of modeling, when compared with the blank group, FDM group showed an increase in the degree of myopia in the right eye, a prolongation of the axial length, a decrease in the content of DA in the retina, and an increase in the expression of MMP-2, TGF-β2 and α-SMA in the sclera. Conversely, the expression of TIMP-2 and TGF-β1 were decreased(P&#x003C;0.01). However, in comparison to the FDM group, the degree of myopia in the 1ng, 5ng, and 10ng groups of guinea pigs decreased, the growth trend of axial length slowed, the content of DA in the retina increased, and the expression of MMP-2, TGF-β2 and α-SMA in the sclera decreased. Furthermore, the expression of TIMP-2 and TGF-β1 in the sclera increased(P&#x003C;0.01). As the concentration of intravitreal injection of VEGF-A165 increased, the degree of myopia in the right eye of guinea pigs gradually increased, and the axial length gradually prolonged. The content of DA in the retina gradually decreased, the expression of MMP-2, TGF-β2, and α-SMA in the sclera gradually increased, while the expression of TIMP-2 and TGF-β1 decreased gradually.CONCLUSION: Intravitreal injection of VEGF-A165 can increase the content of DA in the retina of FDM guinea pigs, affect the expression of MMP-2, TIMP-2, TGF-β1, TGF-β2 and α-SMA in the sclera, and inhibit scleral remodeling of guinea pigs. Notably, the VEGF-A165 at the concentration of 1ng showed the most significant efficacy.

Effects of ICMT gene silencing on the invasion and migration of human salivary adenoid cystic carcinoma cells in vitro / 口腔疾病防治

Objective @#To investigate the effect of isoprene cysteine carboxymethyltransferase (ICMT) gene on the migration and invasion of salivary adenoid cystic cancer cells (SACC) and the related mechanism, to provide experimental evidence for molecular targeted therapy of SACC.@*Methods@# Adenoid cystic cancer cells SACC-LM and SACC-83 were cultured in vitro, and siRNA was transfected into human SACC-LM and SACC-83 cells (experimental group) by transient transfection of a liposome vector. A blank control group and negative control group were set up respectively (transfected NC-siRNA). qRT-PCR was peformed to measure the mRNA expression of ICMT and RhoA in each group after transfection and to determine the silencing efficiency. The expression of ICMT, membrane RhoA, total RhoA, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and Rho associated with coiled helical binding protein kinase 1 (ROCK1) in each group was detected by Western blot. The proliferation abilityies of SACC cells was detected by CCK-8 assay. The migration and invasion ability of SACC cells were detected by comparing the relative healing area of cell scratch assay and the number of Transwell assay cells. @*Results@#After transfection of ICMT-siRNA into SACC-LM and SACC-83 cells, the expression of ICMT gene and protein in the experimental group was significantly decreased compared with the negative control group and blank control group (P<0.05), but there were no significant differences in the expression of RhoA gene and total protein among all groups (P>0.05). The expression of RhoA membrane proteins, ROCK1, MMP-2, MMP-9 in the experimental group was significantly decreased compared with that in the negative control group and blank control group (P<0.05). Cell proliferation ability was significantly decreased (P<0.05). The migration and invasion abilities were significantly decreased (P<0.05). @*Conclusion @#In vitro silencing of ICMT gene can effectively inhibit the migration and invasion of human SACC-LM and SACC-83 cells, and the mechanism may be related to RhoA-ROCK signaling pathway.

miR-184 promotes compensatory lung growth via TIMP-2/MMP-14 in pneumonectomy model / 中国病理生理杂志

AIM:To explore the effect of microRNA-184(miR-184)on compensatory lung growth(CLG)af-ter lobectomy in multiple primary lung cancer(MPLC)and its mechanism.METHODS:(1)Lung tissue samples(n= 16)from MPLC patients and patients with good recovery after lobectomy(CLG)were collected,and the expression of miR-184 was measured by RT-qPCR.(2)Human alveolar epithelial cells were divided into NC-mimic group,miR-184 mimic group,OE-NC group,tissue inhibitor of metalloproteinase-2(TIMP-2)overexpression(OE-TIMP-2)group,and miR-184 mimic+OE-TIMP-2 group according to the transfection(n=3).The expression of miR-184,TIMP-2 mRNA and matrix metalloproteinase-14(MMP-14)mRNA was measured by RT-qPCR,and the protein expression of TIMP-2 and MMP-14 was determined by Western blot.The proliferation of the cells was measured by CCK-8 and colony formation assays.(3)C57BL/6J mice were divided into pneumonectomy(PNX)group and PNX+miR-184 mimic group(n=5).The flexiVent system was used to measure the vital capacity and lung compliance of the mice.Lung volume was measured by water dis-placement method,and lung tissue changes were observed by HE staining.RESULTS:The expression of miR-184 was significantly higher in the patients with better recovery after lobectomy(P＜0.01).Overexpression of miR-184 promoted the proliferation of human alveolar epithelial cells and the recovery of lung function in mice after PNX.In terms of mecha-nism,miR-184 showed targeted binding with TIMP-2,and overexpression of miR-184 promoted the expression of MMP-14 by inhibiting TIMP-2,thereby promoting the proliferation of human alveolar epithelial cells and the recovery of mouse lung function after PNX.CONCLUSION:miR-184 promotes CLG after PNX through the TIMP-2/MMP-14 axis.

Effects of thyroid-stimulating hormone suppressive therapy on programmed death ligand 1 and matrix metalloproteinase-2 expression in thyroid cancer tissue and prognosis / 中国基层医药

Objective:To investigate the effects of thyroid-stimulating hormone (TSH) suppressive therapy on the expression of programmed death ligand 1 (PD-L1) and matrix metalloproteinase 2 (MMP-2) in thyroid cancer tissue and prognosis.Methods:A total of 102 patients with thyroid cancer who underwent surgical resection in Weihai Central Hospital, Qingdao University from April 2016 to April 2018 were included in this study. They were divided into a hormone replacement group and a TSH suppressive therapy group ( n = 51/group). The hormone replacement group was given hormone replacement therapy after surgical resection, and the TSH suppressive therapy group was given TSH suppressive therapy. The expression of PD-L1 and MMP-2 in the pericancerous tissue was compared between the two groups during surgery and 3 and 6 months after surgery. Tumor recurrence and metastasis were compared between the two groups after 6 months, 1 year, and 3 years of follow-up. Results:At 3 and 6 months after surgery, the PD-L1 positive expression rate in the TSH suppressive therapy group was 9.8% (5/51) and 13.7% (7/51), respectively, and the MMP-2 positive expression rate in the TSH suppressive therapy group was 9.8% (5/51) and 13.7% (7/51), respectively, which were significantly lower than 25.5% (13/51), 31.4% (16/51), 27.5% (14/51), and 33.3% (17/51) in the hormone replacement group ( χ2 = 4.32, 5.24, 4.55, 5.45, P = 0.038, 0.022, 0.033, 0.020). At 3 years after surgery, the tumor recurrence and metastasis rate in the TSH suppressive therapy group was 5.9% (3/51), which was significantly lower than 17.6% (10/51) in the hormone replacement group ( χ2 = 4.32, P = 0.038). Conclusion:For patients with thyroid cancer undergoing surgery, TSH suppressive therapy can better inhibit the expression of PD-L1 and MMP-2 in thyroid cancer tissue, reduce the risk of long-term recurrence and metastasis, and have a better clinical application value for improving the prognosis compared with hormone replacement therapy.

Effect of ACE2 on the prognosis of breast cancer and its potential mechanism / 中国药科大学学报

@#This study aims to investigate the effect of transmembrane protein angiotensin converting enzyme 2 (ACE2) on the prognosis of breast cancer and its potential mechanism.Public databases were used to analyze ACE2 expression and its relationship with clinicopathological features and prognosis of breast cancer patients, combined with in vitro experiments to analyze the mechanism of action and immune relevance of ACE2 in breast cancer.Results showed that the expression of ACE2 in breast cancer tissues was significantly lower than that in normal breast tissues, and that its expression was negatively correlated with age, M stage and N1mi stage of breast cancer patients (P < 0.05).Patients with Luminal type breast cancer with high ACE2 expression had poor prognosis, while in the triple-negative breast cancer (TNBC) subtype, ACE2 showed different prognostic significance.In addition, ACE2 is closely associated with the metabolic and immune microenvironment of tumor tissue.In vitro experiments have shown that ACE2 is lowly expressed in MDA-MB-231 cells and may inhibit cell progress by downregulating matrix metalloproteinase 2（MMP2）.The results suggest that the low expression of ACE2 in breast cancer is closely associated with patient prognosis as well as metabolic and immune microenvironment, and that ACE2 may inhibit TNBC cell progress through the MMP2 pathway.

Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats / 解剖学报

Objective To investigate the structural distribution features and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats. Methods Five young SD rats (24 weeks) and five old SD rats (104 weeks) were used,and their cardiac function was examined by echocardiography. Modified Weigert elastic fiber staining, immunohistochemistry, immunofluorescence and Western blotting techniques were used to detect the expression changes of type I and IH collagen fibers and their proteins, elastic fibers and their proteins, matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 2 (TIMP-2), respectively. Results The type I and type IH collagen in the ventricular interstitium of aged rats was very sufficient and wrapped around the cardiomyocytes. Compared with the young rats, the content of collagen protein in the ventricular interstitium of the aged rats significantly increased (P<0. 05). Elastic fibers in the ventricular interstitium of the aged rats were and widely distributed. Compared with the young rats, the number of elastic fibers and the level of elastin in the ventricular interstitium of the aged rats significantly decreased (P<0. 05), and the expression levels of MMP-2 and MMP-9 in ventricular muscle of aged rats increased, and the)' were correlated with the level of elastin. The level of TIMP-2 in ventricular muscle of aged rats decreased with age. Conclusion The number of collagen fibers and elastic fibers in ventricular interstitium of aged rats is fluctuated with each other. With the increase of age, the contents of TIMP-2 and elastic fibers in the ventricular interstitium gradually decreased, and the ratio of collagen fibers to elastic fibers is out of balance.

The role and mechanism of transforming growth factor β1 and matrix metalloproteinase-2 in pancreatic tissue repair and reconstruction in rats with acute pancreatitis / 中华胰腺病杂志

Objective:To investigate the role of transforming growth factor β1 (TGF-β1) and matrix metalloproteinase (MMP)-2 in pancreatic tissue repair and reconstruction in rats with acute pancreatitis and its potential mechanism.Methods:114 male SD rats were randomly divided into normal control group (CON group) and acute edematous pancreatitis model group (AEP group), acute necrotic pancreatitis model group (ANP group), ANP control group and ANP intervention group. The rat AEP model was constructed by subcutaneous injection of caerulein, and the rat ANP model was prepared by intraperitoneal injection of L-arginine. The ANP intervention group and ANP control group were prepared by intraperitoneal injection of TGF-β1 inhibitor SB431542 or DMSO 30 min before, 24 h and 48 h after pancreatitis induction, respectively. Hydroxyproline content in pancreatic tissue was determined by hydroxyproline kit. The expression of TGF-β1, phosphorylated Smad3 (p-Smad3), type Ⅲ collagen and MMP-2 in pancreatic tissue was detected by immunohistochemical method. The activity of MMP-2 was determined by gelatin enzyme spectrometry. The expression levels of MMP-2 and p-Smad3 proteins in pancreatic tissue were detected by Western blot.Results:The hydroxyproline content in CON group was (61.71±8.56)μg/mg protein. The hydroxyproline content in AEP group reached the peak (116.72±8.53)μg/mg on the 3rd day. The peak value of hydroxyproline content in ANP group was (174.93±11.75)μg/mg on day 5. The peak value in ANP group was significantly higher than that in AEP group, and the peak value of hydroxyproline content in AEP group was significantly higher than that in CON group. The hydroxyproline content at day 3, 5 and 7 in the ANP intervention group was (108.07±10.48)μg/mg, (137.14±8.66)μg/mg and (112.35±13.16)μg/mg, respectively, and that at day 3, 5 and 7 in the ANP control group was (132.35±14.2)μg/mg, (175.43±13.75)μg/mg and (137.92±12.65)μg/mg, respectively. TGF-β1 immunohistochemical peak score in control group, AEP group and ANP group was (0.12±0.03), (1.96±0.21) and (3.00±0.28), respectively. p-Smad3 immunohistochemical peak score was (0.15±0.05), (2.05±0.20), and (3.05±0.24), while type Ⅲ collagen immunohistochemical peak score was (0.11±0.04), (1.56±0.15), and (3.10±0.17). MMP-2 immunohistochemical peak score was (0.05±0.03), (1.45±0.20), and (2.45±0.15), respectively. The immunohistochemical peak scores of TGF-β1, p-Smad3, type Ⅲ collagen and MMP-2 in ANP group were significantly higher than those in AEP group. The immunohistochemical peak scores of TGF-β1, p-Smad3, type Ⅲ collagen and MMP-2 in pancreatic tissue of ANP intervention group and ANP control group were (2.36±0.21), (2.25±0.22), (2.47±0.19), (2.00±0.10) and (3.02±0.21), (3.01±0.19), (3.05±0.24), (2.43±0.11), respectively, which in ANP intervention group was significantly lower than those in the ANP control group. The peak value of MMP-2 activity in pancreatic tissue of CON group, AEP group and ANP group was (10.85±1.73), (85.78±7.16) and (115.43±8.7), respectively, which in ANP group was significantly higher than that in AEP group, and in AEP group was significantly higher than that in CON group. In ANP intervention group and ANP control group 3 and 5 days after molding, the expression levels of MMP-2 protein in pancreas were 0.20±0.01, 1.19±0.02, 0.52±0.01, 1.54±0.05, respectively; p-Smad3 protein expression levels were 0.30±0.04, 0.66±0.11, 1.95±0.05, 1.30±0.01, respectively; and MMP-2 and p-Smad3 in ANP intervention group was significantly lower than those the ANP control group. All the differences among the groups above were statistically significant (all P value <0.001). Conclusions:TGF-β1 and MMP-2 play an important role in tissue remodeling and extracellular matrix deposition after acute pancreatitis inflammation.

Tumor microenvironment responsive liposomes blocking CXCL12/CXCR4 pathway and synergistically enhancing immune efficacy of anti-PD-L1 / 药学学报

Blocking immune checkpoint programmed cell death receptor 1 (PD-1) or programmed death receptor-ligand 1 (PD-L1) can enhance anti-tumor activity of effector T cells. However, the lack of response in many patients to PD-1/PD-L1 therapy remains a question. Improving the immunosuppressive tumor microenvironment (TME) to enhance the efficacy of immune checkpoint inhibitors has become a promising cancer treatment strategy. We constructed a liposome system (PD-L1/siCXCL12-Lp) of CXCL12 siRNA and anti-PD-L1 peptide with matrix metalloproteinases (MMPs) responsiveness, which combined the TME regulation of siCXCL12 and the immune regulation of anti-PD-L1 peptide. All animal experiments were approved by the Biomedical Ethics Committee of Peking University. The authors found that PD-L1/siCXCL12-Lp directly down-regulated the expression of CXCL12 in vitro (33.8%) and in vivo (15.5%). It also effectively increased the ratio of CD8+/Treg by 20.0%, which helped the anti-PD-L1 peptide to better exert its immune effect. The combination therapy significantly inhibited tumor growth (52.08%) with great safety, which explored a new idea for cancer immunotherapy.

A review of advances in intraocular fluid detection for high myopia and its relevant fundus diseases / 中华眼底病杂志

The fundus lesions caused by high myopia (HM) often lead to irreversible visual impairment or even blindness. However, the pathogenesis of HM and its fundus lesions is still unclear, the intraocular fluid detection technology of micro samples has brought new prospects for the early diagnosis, monitoring and intervention of the fundus lesions. The molecules associated with HM are various and functionally diverse, intermolecular interactions are staggered and the specific mechanism is complex. With the development of intraocular fluid detection technology, while gradually revealing the role of each molecule in the pathogenesis of HM, it is expected to successfully assist clinical work in the future, providing outpost markers for the progress of myopia and targets for early intervention, or providing a new therapy choice for HM fundus lesions at the molecular level targeting pathogenesis, which is expected to provide more accurate and effective treatment for HM patients in the future.

A review of advances in intraocular fluid detection for high myopia and its relevant fundus diseases / 中华眼底病杂志

The fundus lesions caused by high myopia (HM) often lead to irreversible visual impairment or even blindness. However, the pathogenesis of HM and its fundus lesions is still unclear, the intraocular fluid detection technology of micro samples has brought new prospects for the early diagnosis, monitoring and intervention of the fundus lesions. The molecules associated with HM are various and functionally diverse, intermolecular interactions are staggered and the specific mechanism is complex. With the development of intraocular fluid detection technology, while gradually revealing the role of each molecule in the pathogenesis of HM, it is expected to successfully assist clinical work in the future, providing outpost markers for the progress of myopia and targets for early intervention, or providing a new therapy choice for HM fundus lesions at the molecular level targeting pathogenesis, which is expected to provide more accurate and effective treatment for HM patients in the future.

Advances in the clinical value of tissue inhibitors of metalloproteinase-2 and insulin-like growth factor binding protein 7 in sepsis associated-acute kidney injury / 中华危重病急救医学

Sepsis is an important cause of acute kidney injury (AKI). About 60% of sepsis patients will develop AKI. At present, the standard of clinical diagnosis of AKI is still based on the changes in serum creatinine and urine volume. Because of its lag in time, it may lead to delay in treatment and increase the mortality. To find a new biomarker similar to "troponin" for the diagnosis of AKI, and to achieve the early diagnosis and prevention of AKI, is of great significance to reduce the mortality of AKI. In recent years, it has been found that tissue inhibitors of metalloproteinase-2 (TIMP-2) and insulin-like growth factor-binding protein 7 (IGFBP7) can be used for early diagnosis of sepsis associated-acute kidney injury (SA-AKI). They also have important values in risk stratification, prognosis judgment, intervention and other aspects of SA-AKI. In this paper, the research progress of the application of TIMP-2 and IGFBP7 in SA-AKI is reviewed.

Agkistrodon halys venom antitumor component-I inhibits vasculogenic mimicry in triple-negative breast cancer cells in vitro by down-regulating MMP2 / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of agkistrodon halys venom antitumor component-I (AHVAC-I) on vasculogenic mimicry (VM) formation in triple-negative breast cancer MDA-MB-231 cells and explore its possible mechanism.@*METHODS@#CCK8 assay was used to determine the optimal concentration of AHVAC-I for cell treatment based on its halfinhibitory concentration (IC50). MDA-MB-231 cells were treated with different concentrations of AHVAC-I or 5-Fu, and the changes in vasomimetic capacity of the cells were examined using Matrigel assay. The expression levels of matrix metalloproteinase-2 (MMP2) and MMP9 in the treated cells were detected using quantitative PCR and Western blotting.@*RESULTS@#Compared with the control treatment with culture medium, treatment with 5, 10 and 20 μg/mL AHVAC-I significantly reduced vasomimetic ability of MDA-MB-231 cells in a dose-dependent manner (P < 0.01). MMP2 supplementation obviously restored the vasomimetic ability of the cells inhibited by AHVAC-I.@*CONCLUSION@#AHVAC-I inhibits VM formation in triplenegative breast cancer cells in vitro by down-regulating MMP2 production.

Changes of biomarkers in fluid derived from peritoneal cavity and their relationship with transport rate in peritoneum in patients with new peritoneal catheter / 国际生物医学工程杂志

Objective:To investigate the changes of biomarkers in peritoneal dialysis patients' peritoneal drainage fluid and their relationship with the peritoneal small molecule solute transport rate (PSTR).Methods:Seventy newly-tubed peritoneal dialysis patients from the Peritoneal Dialysis Center of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from September 29, 2014 to April 26, 2018 were selected. The levels of biomarkers plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-2 (MMP-2) and vascular endothelial growth factor (VEGF) in the peritoneal dialysis priming fluid were measured at different time points and 4 h dialysate/blood muscle (D/P) creatinine values at 2 years of follow-up, and the correlation between biomarkers in the extracted peritoneal fluid and 4 h D/P creatinine was examined.Results:Longitudinal studies showed an increase in PAI-1 ( P<0.001) and VEGF ( P=0.04) with increasing duration of peritoneal dialysis. PSTR levels at baseline and after 2 years of follow-up were significantly correlated with PAI-1, MMP-2, and VEGF levels at baseline. PSTR at 2 years was also correlated with MMP-2 levels at 6 months and PAI-1 levels at baseline. Conclusions:The biomarkers PAI-1, MMP-2, and VEGF in peritoneal dialysis drainage fluid are positively correlated with PSTR in peritoneal dialysis patients during the 2-year period.

Effect of microRNA-221 targeting tissue inhibitor of metalloproteinase-2 to mediate Akt / mTOR signaling pathway on non-small cell lung cancer transplanted tumor mouse model / 解剖学报

Objective To explore the effects of miR-221 on tumor cell proliferation, migration and invasion in nonsmall cell lung cancer (NSCLC) xenograft model mice, and to preliminarily analyze its possible mechanism of regulating Akt/ mammalian target of rapamycin(mTOR) signaling pathway by targeting tissue inhibitor of metalloproteinase-2 (TIMP-2) on tumor cells in non-small cell lung cancer (NSCLC) through tumor-bearing nude mice. Methods The A549 cells were divided into control group, mimic group, TIMP-2 group and mimic+TIMP-2 group. The mimic group and TIMP-2 group were transfected with miR-221 mimic and TIMP-2 overexpression plasmids, respectively. The mimic + TIMP-2 group was simultaneously transfected with miR-221 mimic and TIMP-2 overexpression plasmids. The control group was transfected with the same amount of negative control plasmid. After transfection, the cells of each group were injected subcutaneously into the left forelimb to construct the corresponding 4 groups of NSCLC mouse models. The proliferation-related protein (Ki67) was detected by immunohistochemical staining to detected the effect of cell proliferation ability. Matrix metalloproteinase-2 (MMP-2) and N-cadherin proteins in each group were tested by Western blotting to assess and compare the abilities of migration and invasion. The levels of miR-221, TIMP-2 and Akt/ mTOR pathways in bone marrow and tumor tissues were detected by Real-time PCR and Western blotting. Results When co-transfected with wild type(WT)-TIMP-2 and miR-221 mimic, the relative luciferase activity in the cells reduced significantly (P<0. 05). Compared with the control group, the tumor mass, volume, Ki67, MMP-2 and N-cadherin protein expression levels, miR-221 and Akt/ mTOR pathway levels were increased significantly, while the levels of TIMP-2 mRNA and protein were significantly reduced in the mimic group (P<0. 05). Compared with the control group, the levels of TIMP-2 mRNA and protein in the TIMP-2 group increased significantly, while the other indicators decreased significantly (P<0. 05). Tumor tissue mass, volume, Ki67, MMP-2, Ncadherin, miR-221 and Akt/ mTOR pathway levels in mimic+TIMP-2 group were significantly lower than those in the mimic group and significantly higher than those in the TIMP-2 group, while TIMP-2 mRNA and protein levels were significantly higher than those in the mimic group and significantly lower than those in the TIMP-2 group (P<0. 05). Conclusion In the NSCLC transplanted tumor mouse model, miR-221 may mediate the Akt/ mTOR pathway by targeting the expression of TIMP-2 protein to promote cell proliferation, migration and invasion.

Nitric oxide regulates adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells

Anoikis is a type of apoptosis that occurs in response to the loss of adhesion to the extracellular matrix (ECM). Anoikis resistance is a critical mechanism in cancer and contributes to tumor metastasis. Nitric oxide (NO) is frequently upregulated in the tumor area and is considered an important player in cancer metastasis. The aim of this study was to evaluate the effect of NO on adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells. Here, we report that anoikis-resistant endothelial cells overexpress endothelial nitric oxide synthase. The inhibition of NO release in anoikis-resistant endothelial cells was able to decrease adhesiveness to fibronectin, laminin, and collagen IV. This was accompanied by an increase in cell invasiveness and migration. Furthermore, anoikis-resistant cell lines displayed a decrease in fibronectin and collagen IV protein expression after L-NAME treatment. These alterations in adhesiveness and invasiveness were the consequence of MMP-2 up-regulation observed after NO release inhibition. The decrease in NO levels was able to down-regulate the activating transcription factor 3 (ATF3) protein expression. ATF3 represses MMP-2 gene expression by antagonizing p53-dependent trans-activation of the MMP-2 promoter. We speculate that the increased release of NO by anoikis-resistant endothelial cells acted as a response to restrict the MMP-2 action, interfering in MMP-2 gene expression via ATF3 regulation. The up-regulation of nitric oxide by anoikis-resistant endothelial cells is an important response to restrict tumorigenic behavior. Without this mechanism, invasiveness and migration potential would be even higher, as shown after L-NAME treatment.

Study of plasma matrix metalloproteinase-2 and matrix metalloproteinase-9 level during concurrent chemoradiotherapy and clinicopathologic significance in patients with nasopharyngeal carcinoma / 中国医师杂志

Objective:To investigate the changes of the plasma matrix metalloproteinase (MMP)-2 and MMP-9 levels and their clinical significances during the course of concurrent chemoradiotherapy (CCRT) in nasopharyngeal carcinoma (NPC) patients.Methods:From January 2018 to June 2019, 46 patients with nasopharyngeal carcinoma were treated in the department of oncology, Changsha Central Hospital Affiliated to Nanhua University. All patients were confirmed by pathology. They were divided into early NPC group ( n=32) and invasive NPC group ( n=16) according to the degree of invasion. The early NPC group was treated with concurrent chemoradiotherapy alone, and the invasive NPC group was treated with neoadjuvant chemotherapy combined with concurrent chemoradiotherapy. Blood samples were collected at four stages of the treatment, and the concentrations of MMP-2 and MMP-9 were detected by enzyme linked immunosorbent assay (ELISA). Results:The longer the treatment time, the lower the concentration of MMP-9 ( P=0.007) in early NPC group; There was no significant difference in MMP-9 level before treatment, after neoadjuvant chemotherapy, after concurrent chemoradiotherapy, at the end of treatment and the first follow-up ( P>0.05) in invasive NPC group. There was no significant difference in the content of MMP-2 between the two groups before and after treatment ( P>0.05). There was no correlation between serum MMP-2 and MMP-9 levels and tumor stage, lymph node metastasis, tumor invasion and response rate ( P>0.05) in invasive NPC patients, while the level of MMP-9 was positively correlated with white blood count (WBC) and neutrophil count ( r=0.85, P=0.004, r=0.82, P=0.003); The ratio of MMP-9/MMP-2 was positively correlated with WBC and neutrophil count ( r=0.86, P=0.003, r=0.83, P=0.001). Conclusions:Synchronous radiotherapy and chemotherapy can reduce the serum MMP-9 level in early stage NPC patients, but it has no effect on the serum MMP-9 level in patients with invasive NPC, which suggests that synchronous radiotherapy and chemotherapy can not prevent the proliferation and distant metastasis of cancer cells in patients with invasive NPC.

The predictive value of TIMP-2 and IGFBP7 in delayed renal function recovery after DCD renal transplantation / 中华泌尿外科杂志

Objective:This study is to investigate the predictive value of serum levels of TIMP-2 and insulin-like growth factor-binding protein 7(IGFBP7) in patients with DCD(donation after cardiac death) kidney transplantation.Methods:A prospective research design was used to select DCD kidney transplant patients admitted to the Li Huili Hospital of Ningbo University from January 2018 to October 2020.Inclusion criteria: ①Complete data; ②There were no serious complications affecting the function of the transplanted kidney in the early postoperative period.Exclusion criteria: ①Incomplete data; ②Patients were unable or unwilling to cooperate with the study; ③Severe complications affecting the function of the transplanted kidney occurred early after the operation.The ELASE method was used to quantitatively detect the serum TIMP-2 and IGFBP7 levels at 6, 12, 24, 48, 72 hours and 7 days after renal transplantation, and monitor the serum creatinine values during the same period and 21 days after the operation. According to the occurrence of DGF, the measured values of TIMP-2 and IGFBP7 at different time points and their product's ability to predict the occurrence of DGF after kidney transplantation were analyzed. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic efficacy of TIMP-2 and IGFBP7 for DGF.Results:A total of 33 patients were enrolled, 7 patients (21.2%) in the DGF group and 26 patients (78.8%) in the non-DGF group. Between the two groups, the donor glomerular filtration rate were [98.5(15.8-132.5)ml/(min·1.73m 2) and 79.1(60.6-102.5)ml/(min·1.73m 2)], recipient gender (male/female: 3/4 cases and 10/16 cases), recipient age [48(34-56) Years old and 45(23-61) years old], the recipient's preoperative creatinine [1114.0(731.4-1293.0)μmol/L and 858.4(657.6-1051.9)μmol/L], the recipient's preoperative urea nitrogen [15.0(13.2-19.6)mmol/L and 17.3(13.6-20.9)mmol/L], receptor preoperative albumin [43.5(38.5-45.3)mmol/L and 41.2(37.5-46.1) mmol/L], recipient dialysis method [hemodialysis/peritoneal dialysis: 3/4 cases and 9/17 cases], warm ischemia time [6(5-7) and 5(4-6) min, there was no statistically significant difference] ( P>0.05). The values of serum IGFBP7 and TIMP-2×IGFBP7 in the DGF group were higher than those in the non-DGF group at all time points ( F=15.753, P=0.040; F=13.000, P=0.024), while serum TIMP-2 was not significant between the two groups difference ( F=1.157, P=0.075). For the diagnostic value of DGF, the AUC of serum IGFBP7 at 48 h after surgery was 0.863 (95% CI 0.696-1.000, P=0.004). When 5.97 ng/ml was used as the cut-off value, the sensitivity was 85.7% and the specificity was 80.8 %. The AUC of TIMP-2×IGFBP7 at 48 hours after surgery was 0.819 (95% CI 0.641-0.996, P=0.011). When 62.06(ng/ml) 2 was used as the cutoff value, the sensitivity was 71.4% and the specificity was 80.8%.There was no statistical difference in the area under the curve between the two ( P>0.05). There were differences in the dynamic trend of serum IGFBP7 and creatinine in the DGF group. Serum IGFBP7 at 7 days after surgery was positively correlated with creatinine at 21 days after surgery. Conclusion:Serum IGFBP7 and TIMP-2×IGFBP7 could predict the occurrence of DGF after DCD donor kidney surgery. The predictive value changes with time. Among them, 48h and 7d after surgery are the most valuable. However, serum TIMP-2 has not been found to have predictive value in this study.