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>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<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<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.

Mechanism of rhynchophylline solid lipid nanoparticles inhibiting the proliferation of airway smooth muscle cells in asthmatic model mice / 中国药房

OBJECTIVE To study the inhibitory effect mechanism of rhynchophylline solid lipid nanoparticles （Rhy-SLN） on the proliferation of airway smooth muscle cells （ASMCs） in asthmatic model mice. METHODS Asthma model was prepared by ovalbumin+calmogastrin sensitization. The primary isolation and culture of ASMCs were performed， and morphological observation and identification were also conducted [when α -smooth muscle actin （α -SMA） appeared red and Desmin appeared green in ASMCs， indicating successful cultivation of ASMCs]. The cells were divided into blank group （ASMCs of normal mice）， model group （ASMCs of asthma model mice）， Rhy-SLN group （ASMCs of asthma model mice）， recombinant suppressors of cytokine signaling 1 （SOCS1） overexpression group （ASMCs of asthma model mice transfected with SOCS1 vector）， SOCS1-RNAi group （ASMCs of asthma model mice transfected with SOCS1-RNAi vector） and SB203580 group [p38 mitogen-activated protein kinase （p38 MAPK） inhibitor， ASMCs of asthma model mice]. The cells of each group were added into the corresponding culture medium containing drug （10 μmol/L） or not containing drug for 24 hours. MTT method was used to detect the proliferation of ASMCs in asthmatic mice； Western blot assay was used to detect the protein expressions of α-SMA， interleukin-1β （IL-1β）， SOCS1， p38 MAPK and phosphorylated p38 MAPK （p-p38 MAPK） in ASMCs. RESULTS The primary ASMCs of mice varied in shape and size， presenting irregular， spindle and triangular shapes；α-SMA appeared red and Desmin appeared green， indicating successful cultivation of ASMCs. Compared with model group， ASMCs absorbance values and protein expressions of α -SMA， p38 MAPK， and p-p38 MAPK were reduced significantly in Rhy- SLN group， SOCS1 overexpression group and SB203580 E-mail：wangmeng106@163.com group， while protein expression of SOCS1 （except for group） was increased significantly （P＜0.05）； protein expressions of IL-1β was reduced significantly in ASMCs （P＜ 0.05）. ASMCs absorbance values and protein expressions of α-SMA， SOCS1， p38 MAPK and p-p38 MAPK were increased significantly in SOCS1-RNAi group （P＜0.05）. CONCLUSIONS Rhy-SLN can inhibit the proliferation of ASMCs， the mechanism of which may be associated with overexpression of SOCS1 and inhibiting the protein expressions of IL-1β and p38 MAPK.

Effect of mycophenolate mofetil alleviates carbon tetrachloride-induced liver fibrosis in mice / 中南大学学报(医学版)

OBJECTIVES@#Hepatic fibrosis is a serious pathological consequence of chronic liver disease. Mycophenolate mofetil (MMF) is a commonly used immunosuppressant after organ transplant. However, the relationship between MMF and hepatic fibrosis remains unclear. This study aims to explore the effect of MMF on hepatic fibrosis in mice and the potential mechanism.@*METHODS@#A total of 24 mice (male, 8-week old, C57BL/6) were randomly divided into a control group, a MMF group, a carbon tetrachloride (CCl4) group and a CCl4+MMF group (n=6 in each group). After the mice were sacrificed, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected. The liver tissues were taken up for Masson staining and collagen I (COL1) immunohistochemistry. The levels of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) were detected by Western blotting. Finally, the levels of mRNA for TGF-β1, α-SMA, and COL1 were detected using real-time PCR.@*RESULTS@#Compared with the CCl4 group, the ALT and AST levels were lower (both P<0.05), the degree of liver fibrosis was alleviated, and the deposition of COL1 in the liver was significantly decreased (P<0.01) in the CCl4+MMF group. Compared with the CCl4 group, the protein expression levels of TGF-β1 and α-SMA were significantly decreased (both P<0.05) and the relative expression levels of TGF-β1, α-SMA and COL1 mRNA in the liver were significantly decreased (all P<0.05) in the CCl4+MMF.@*CONCLUSIONS@#MMF could reduce CCl4-induced hepatic fibrosis, which might be related to the inhibition of TGF-β1. This study is expected to provide a target for the treatment of hepatic fibrosis.

Primary leiomyosarcoma of ovary: A rare malignancy as an incidental finding

Primary leiomyosarcoma (PLMS) of the ovary is extremely rare tumors comprising 1% of ovarian tumors. About 3% of all ovarian malignancies are primary ovarian sarcomas. Only 72 cases have been reported till date. A 57-year-old postmenopausal female presented with abdominal pain for the last 6 months. Ultrasonography and MRI revealed a heterogeneously enhancing solid lobulated mass in the left adnexa abutting the fundus of the uterus and bowel loops. The endometrial cavity was normal. Ovarian markers CA 125, CEA, CA 19.9, and all hematological parameters were within normal limits. LDH was near normal (284 IU/ml). The specimen was sent for frozen section and a diagnosis of malignant spindle cell lesion of ovary was rendered. Histopathology of the ovarian mass revealed intersecting fascicles of tumor cells consisting of ovoid to spindle-shaped cells having a moderate amount of cytoplasm. Bizarre and atypical cells were seen singly dispersed and in small aggregates along with the brisk mitotic activity. Focal areas of necrosis and hemorrhage were also noted. Immunohistochemistry showed strong positivity for smooth muscle actin and Caldesmon while focal positivity for Desmin and Epithelial Membrane Antigen (EMA) was noted. The lesion was negative for Inhibin, Calretinin, and CD 117 and S100. The final diagnosis of primary ovarian Leiomyosarcoma was given based on histopathology and Immunohistochemistry. PLMS of the ovary are rare incidental findings in postmenopausal women. These are highly malignant tumors and carry a poor prognosis. Hence, early diagnosis and surgical treatment with cytoreduction improve patient survival.

Effect of Morindae Officinalis Radix Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Improvement of Renal Function and HPG Axis of Kidney Yang Deficiency Model Rats Induced by Adenine / 中国实验方剂学杂志

ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.

Effect of Morindae Officinalis Radix Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Improvement of Renal Function and HPG Axis of Kidney Yang Deficiency Model Rats Induced by Adenine / 中国实验方剂学杂志

ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.

Effect and mechanism of terminal fucosylation inhibitor on ciclosporin-induced renal epithelial-mesenchymal transition / 器官移植

Objective To evaluate the effect and mechanism of terminal fucosylation inhibitor 2-deoxy-D-galactose (2-D-gal) on ciclosporin (CsA)-induced renal epithelial-mesenchymal transition (EMT). Methods Fifteen male C57BL/6 mice aged 8-10 weeks were randomly and evenly divided into the control group (Ctrl group), CsA group and CsA+2-D-gal group (n=5). The expression levels of fucosyltransferase 1 (FUT1), EMT-associated proteins including E-cadherin, Vimentin, α-smooth muscle actin (α-SMA) in the kidney tissues of the Ctrl and CsA groups were detected by Western blot. The expression levels of terminal fucose in the kidney tissues of Ctrl and CsA groups were determined by immunofluorescence. The renal fibrosis of mice in each group was evaluated by Masson staining. The blood urea nitrogen and serum creatinine levels of mice in each group were detected. The in vitro EMT model of renal tubular epithelial cell HK2 was induced by CsA. HK2 cells were stimulated with 0, 2.5, 5.0 and 10.0 μmol/L CsA for 24 h, respectively. In addition, HK2 cells were divided into the Ctrl, 2-D-gal, CsA and CsA+2-D-gal groups. The morphology of HK2 cells after stimulation with different concentrations of CsA and in each group was observed. The expression levels of FUT1, E-cadherin, Vimentin and α-SMA in HK2 cells after stimulation with different concentrations of CsA and in each group were detected by Western blot. The expression level of terminal fucose in HK2 cells of the Ctrl and CsA groups was measured by immunofluorescence. Results Compared with the Ctrl group, the relative expression of E-cadherin protein was down-regulated, those of FUT1, Vimentin and α-SMA proteins were up-regulated (all P < 0.05), and that of terminal fucose in the mouse kidney tissues was up-regulated in the CsA group. Compared with the Ctrl group, the blood urea nitrogen and serum creatinine levels in the CsA and CsA+2-D-gal groups were up-regulated (all P < 0.05). Compared with the CsA group, the blood urea nitrogen and serum creatinine levels in the CsA+2-D-gal group were down-regulated (both P < 0.05). Compared with the Ctrl group, the collagen fiber deposition was increased and the relative expression of α-SMA protein was up-regulated in the mouse kidney tissues of CsA and CsA+2-D-gal groups (all P < 0.05). Compared with the CsA group, the collagen fiber deposition was decreased and the relative expression of α-SMA protein in the mouse kidney tissues was down-regulated in the CsA+2-D-gal group (both P < 0.05). With the increase of CsA concentration, the morphology of HK2 cells gradually became longer and thinner from original normal cobblestone shape, the relative expression levels of FUT1, Vimentin and α-SMA protein in HK2 cells were up-regulated, and that of E-cadherin protein was down-regulated in a concentration-dependent manner. Compared with the Ctrl group, the expression level of terminal fucose of HK2 cells was up-regulated in the CsA group. After CsA treatment combined with 2-D-gal intervention, the morphology of HK2 cells in the CsA+2-D-gal group was restored to resemble that of normal HK2 cells. Compared with the CsA group, the relative expression of E-cadherin protein in HK2 cells was up-regulated, whereas those of Vimentin and α-SMA proteins were down-regulated in the CsA+2-D-gal group (all P < 0.05). Conclusions CsA may induce EMT both in vivo and in vitro, and the terminal fucosylation is increased. 2-D-gal may inhibit CsA-induced EMT by suppressing the terminal fucosylation.

Role of three drugs in traumatic proliferative vitreoretinopathy in rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effect of triamcinolone acetonide(TA), artesunate(ART), and luteolin(LU)on the prevention and treatment of traumatic proliferative vitreoretinopathy(TPVR). <p>METHODS: Forty-eight cyanotic blue rabbits were selected to prepare TPVR animal models by making a penetrating eye injury and intravitreal injection of 0.3mL platelet-rich plasma, and were randomly divided into four groups(<i>n</i>=12), in which the vitreous cavity of the control group was injected with 0.1mL saline; The vitreous cavity of the TA group was injected with 0.1mL(1mg/mL)triamcinolone acetonide; The vitreous cavity of the ART group was injected with 0.1mL(20μg/mL)artesunate; 0.1mL(10μg/mL)luteolin was injected into the vitreous cavity of the LU group. The vitreous and retinal proliferation were observed by fundus photography and ocular ultrasound at 1, 2, 3 and 4wk postoperatively. The expression levels of α-SMA and VIM protein in the vitreous fluid of each group of rabbit eyes were detected by Western Blot at 28d postoperatively, and the retinal tissue structure of each group was observed by retinal HE staining. <p>RESULTS: At 28d postoperatively, the TPVR grading of rabbit eyes in the TA, ART and LU groups were significantly lower than that in the control group(<i>P</i><0.05), and the TPVR grading of rabbit eyes in the TA group was significantly lower than that in the ART and LU groups(<i>P</i><0.05). The expression levels of α-SMA and VIM proteins in the vitreous fluid of the rabbit eyes in the TA, ART and LU groups were significantly lower than those in the control group at 28d after surgery(<i>P</i><0.01). The results of HE staining showed that the arrangement of retinal layers in rabbit eyesin the control group were disordered, severely distorted or locally broken, the structure of each layer were unclear, the anterior membrane was obviously thickened, and the retina was obviously detached; The arrangement of retinal layersin rabbit eyes in the LU group were slightly distorted, inflammatory exudation was visible in front of the retina, and the retina was superficially detached; The structure of retina in rabbit eyes in the ART group were clear, with mild edema and superficial detachment; The structure of retinal layers in rabbit eyes in the TA group were clear, the arrangement was still neat, the retinal folds were locally visible, and there was no retinal detachment.<p>CONCLUSION: Intravitreal injection of triamcinolone acetonide, artesunate and luteolin were all effective in preventing and treating traumatic TPVR, among which triamcinolone acetonide has the most obvious effect.

The value of acidic mammalian chitinase in early diagnosis of occupational pneumoconiosis / 中国职业医学

OBJECTIVE: To observe the expression of acidic mammalian chitinase(AMCase) in lung tissue of silicosis model rats, and bronchoalveolar lavage fluid(BALF) and serum of patients with occupational pneumoconiosis, and to evaluate the value of AMCase in the early diagnosis of pneumoconiosis. METHODS: i) The specific pathogen free adult male Wistar rats were randomly divided into control group and model group, with 15 rats in each group. The rats in the silicosis model group was exposed to free silica dust with a concentration of 2 000.0 mg/m~3 by dynamic inhalation for three hours a day, while the rats in control group were not exposed to dust. Five rats in the two groups were sacrificed at 4, 12 and 24 weeks after dust exposure. ii) By random number table method, a total of 191 patients with occupational pneumoconiosis who received large capacity lung lavage were selected as the pneumoconiosis group, 12 dust-exposed workers who received large capacity lung lavage were selected as the dust control group, and 200 healthy coal miners exposed to dust were selected as healthy control group. iii) Western blotting was used to detect the relative protein expression of AMCase, type Ⅰ collagen(COLⅠ), α-smooth muscle actin(α-SMA) in lung tissues of the rats and the relative protein expression of AMCase in human BALF. Enzyme linked immunosorbent assay was used to detect the level of AMCase protein in human serum. Receiver operating characteristic(ROC) curve was used to evaluate the value of AMCase protein level in human serum for early diagnosis of pneumoconiosis. RESULTS: The relative expression of AMCase, COLⅠand α-SMA protein in lung tissue of rats in the silicosis model group were higher than that of control group(all P<0.01). The relative expression of AMCase protein in BALF of pneumoconiosis group and pneumoconiosis stage Ⅰ, Ⅱ and Ⅲ subgroups were higher than that of dust control group(all P<0.05). The level of AMCase protein in serum of pneumoconiosis group and pneumoconiosis stage Ⅰ, Ⅱ and Ⅲ subgroups were higher than that of healthy control group(all P<0.05). The results of ROC curve analysis showed that the area under ROC curve was 0.78(95% confidence interval: 0.74-0.82).When the cut-off value of serum AMCase protein level was 466.0 ng/L, the sensitivity was 73.8%, and the specificity was 72.6%. CONCLUSION: AMCase protein in human serum has value for early diagnosis of pneumoconiosis and it could be a potential biomarker for early diagnosis of pneumoconiosis.

Monotypic angiomyolipoma of the nasal cavity: An extremely rare cause of nasal mass with recurrent epistaxis

Monotypic angiomyolipoma is usually found in the kidneys and is composed predominantly of epithelioid cells which show positivity for melanocyte and smooth muscle markers. It can pose a diagnostic challenge due to a range of differential diagnosis. We report the second case of monotypic angiomyolipoma of nasal cavity and first from India in a 54-year-old male who presented with a nasal polyp. Grossly the tumor was well circumscribed and un-encapsulated. Microscopy showed a large number of epithelioid cells mixed with a few spindle cells, varying sized blood vessels, and focal areas of adipose tissue. Immunohistochemistry was positive for smooth muscle actin (SMA) and human melanoma black (HMB-45) stains. It is important to identify this tumor as it can sometimes be mistaken for malignancy and only needs endoscopic resection.

Therapeutic effect of human adipose stem cells derived exosomes on carbon tetrachloride induced liver fibrosis in rats / 中国组织工程研究

BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets.OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.

Mechanism of leflunomide in regulating pulmonary fibrosis by regulating miR-449a / 药学实践杂志

Objective To investigate the mechanism of leflunomide (LEF) in regulating pulmonary fibrosis by regulating microRNA (miR)-449a. Methods Human lung fibroblasts MRC-5 were divided into 6 groups: control group, LEF group, LEF+mimic group, mimic group, LEF+inhibitor group and inhibitor group. MiR-449a was overexpressed or silenced by plasmid transfection with miR-449a mimic or inhibitor and ncubate for 48 h at 5 mg / L LEF. The cell viability, cell proliferation ability and apoptotic rate of each group were measured by CCK-8 method, clone formation experiment and flow cytometry. Immunofluorescent staining was used to detect α smooth muscle actin (α-SMA) and collagen I (col I). The levels of miRNA and protein were detected using qPCR and Western blot, respectively. Results The miR-449a level in the mimic group was significantly higher than that in the control group (P<0.05). The level of miR-449a in LEF group and inhibitor group was significantly lower than that in control group (P<0.05). The expression level of miR-449a in LEF+mimic group was significantly higher than that in LEF group, and the level of miR-449a in LEF+inhibitor group was significantly lower than that in LEF group (P<0.05). The cell viability and cell proliferation ability of the LEF group and inhibitor group were significantly higher than those of the control group (P<0.05). The cell viability and cell proliferation ability of the mimic group were significantly lower than those of the control group (P<0.05). The cell viability and cell proliferation ability of the LEF+mimic group were significantly lower than those of the LEF group, while the cell viability of the LEF+inhibitor group was significantly higher than that of the LEF group (P<0.05). The apoptosis rate of LEF group and inhibitor group was lower than that of control group (P<0.05). The apoptosis rate of mimic group was significantly higher than that of control group (P<0.05). The apoptosis rate of LEF+mimic group was significantly higher than that of LEF group, while the apoptosis rate of LEF+inhibitor group was significantly lower than that of LEF group (P<0.05). The fluorescence intensity of α-SMA and Col I proteins in LEF group and inhibitor group were significantly higher than those in control group (P<0.05). The relative fluorescence intensity of mimic group was lower than that of control group (P<0.05). The relative fluorescence intensities of α-SMA and Col I proteins in LEF+mimic group were significantly lower than those in LEF group, while the relative fluorescence intensities of α-SMA and Col I protein in LEF+inhibitor group were significantly higher than those in LEF group (P<0.05). The levels of p-JNK / JNK in LEF group and inhibitor group were higher than those in control group (P<0.05). The p-JNK / JNK level in the mimic group was significantly lower than that in the control group (P<0.05). The level of p-JNK / JNK in LEF+mimic group was significantly lower than that in LEF group, while the level of p-JNK / JNK in LEF+inhibitor group was significantly higher than that in LEF group (P<0.05). Conclusion LEF may activate the JNK pathway by inhibiting the expression of miR-449a in lung fibroblasts, thereby inducing fibroblast activation and proliferation, inhibiting apoptosis, and causing pulmonary fibrosis.

Anti-scarring effect of rapamycin following filtering surgery in rabbit eyes / 南方医科大学学报

OBJECTIVE@#To study the effect of rapamycin on scar formation in rabbit eyes following filtering operation and explore the possible mechanism.@*METHODS@#Ninety-six healthy adult rabbits were subjected to trabeculectomy of the left eye and subsequently randomly divided into 4 groups (=24) for treatment with castor oil (control) or rapamycin (1%, 3%, or 5%) eye drops of the operated eyes 4 times a day. The morphology and function of the filtering blebs of the rabbits were compared at 7, 14, 21 and 28 days after the operation; at each of the time points, 6 rabbits from each group were euthanized for detection of expressions of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) in the tissues in the surgical area using immunohistochemistry. Cultured rabbit subconjunctival fibroblasts (RTFSs) were treated with different concentrations of rapamycin (0.06, 0.25, 1, and 4 mg/L) and the cell apoptosis was detected using flow cytometry.@*RESULTS@#In the first, second and third weeks after the operation, the rate of functional follicle formation was significantly higher in the 3 rapamycin groups than in the control group ( < 0.05), and the number of α- SMA-positive fibroblasts decreased over time in the 3 rapamycin groups. In cultured RTFSs, treatment with rapamycin at different concentrations resulted in increased apoptosis of the cells, and rapamycin above 0.25 mg/L significantly increased the cell apoptosis in a dose-dependent manner.@*CONCLUSIONS@#Rapamycin can inhibit hyperplasia of the filtering passage tissue, helps to preserve the functional filtering blebs and prolong their life span, and induces apoptosis of RTFS.

IL-17A activates mouse lung fibroblasts through promoting chemokine CXCL12 secretion / 浙江大学学报·医学版

OBJECTIVE@#To investigate the role of IL-17A in promoting the activation of lung fibroblasts and the secretion of chemokine CXCL12, and to explore the possible mechanism.@*METHODS@#Lung tissues of BALB/c mice were collected after intraperitoneal injection of recombinant mouse IL-17A (rmIL-17A). Real-time RT-PCR and Western blotting were used to detect the expression levels of α-smooth muscle actin (α-SMA) and collagen I in lung tissues, and immunohistochemical staining and real-time RT-PCR were used to determine the expression of CXCL12. Normal mouse primary lung fibroblasts were isolated and cultured, and identified by immunofluorescence staining with optical microscopy. Cells and supernatant of culture medium were collected after stimulation with rmIL-17A at different concentrations. mRNA levels of α-SMA, collagen I, and CXCL12 in the cells were determined by real-time RT-PCR, and the levels of collagen I and CXCL12 in the supernatant of culture medium were determined by ELISA.@*RESULTS@#The mRNA and protein levels of α-SMA and collagen I in the lung tissue of mice injected with rmIL-17A were significantly increased compared with the control group (all @*CONCLUSIONS@#s: IL-17A can promote the activation of lung fibroblasts and translation into myofibroblast. The secretion of collagen is increased, which promote the deposition of extracullular matrix, and leads to the occurrence and development of lung fibrosis. CXCL12, a chemokine secreted by activated fibroblasts, may be involved in this process.

Effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma Extracts on Expression of α-SMA and Runx2 Protein After High Glucose-induced Vascular aging in Mice / 中国实验方剂学杂志

Objective::To investigate the effects of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts (GNC) on the protein expression of α-smooth muscle actin (α-SMA) and runt-related transcription factor2(Runx2) after high glucose-induced vascular aging in mice, and elucidate the protective mechanism of GNC in delaying vascular aging. Method::Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin (STZ). After successful modeling, the mice received high-fat diet for 7 months, and then they were randomly divided into model group, GNC low-dose and high-dose groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). The drug was given by intragastric administration once a day for 9 weeks. Seven days before tissues collection, a new batch of 4-week-old male C57BL/6 mice were purchased and fed normally for 1 week as a youth group. The general condition of the mice was observed. Morphological changes of the common carotid artery in mice were determined by hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM). Masson trichromatic staining was used to observe the fibrosis of common carotid artery in mice. The expression levels of matrix metalloproteinases-2 (MMP-2), cyclin-dependent kinase inhibitor 2A (p16), cyclic-dependent kinase inhibitor 1A (p21), α-SMA and Runx2 in the common carotid arteries of mice were detected by immunohistochemistry. Result::The results of HE, TEM and Masson showed that there was almost no change in the inimal and adventitial thickness, ultrastructure and relative contents of collagen and elastic fibers in the common carotid arteries of mice between the youth group and normal control group. As compared with the normal control group, the intima of the common carotid artery in the model group was not smooth, the endothelial cells were almost completely detached, the cytoplasm was lysed, the inner elastic membrane became thinner, fractured, or even detached, and the proliferating collagen fibers sneaked into the tunica media. The hyperplasia of tunica media and tunica adventitia was obvious and disordered (P<0.01). The vascular smooth muscle cells showed deformations, protuberances, bifurcations, and even fragmentation, and focal necrosis was observed. There were significantly more vacuoles, lysosomes, and obvious autophagy vesicles. The relative content of collagen and elastic fibers in vascular walls increased significantly (P<0.01). Compared with the model group, the above situation was relieved in each administration group (P<0.01). The results of immunohistochemistry showed that high glucose induced high expression of MMP-2, p16, p21 and Runx2 in the common carotid arteries(P<0.01), low expression of α-SMA(P<0.01), and the protein expression tended to be normal after drug intervention(P<0.05, P<0.01). Conclusion::High glucose can induce the aging of common carotid artery in mice and change the expression of α-SMA and Runx2 proteins. The Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can delay vascular aging by regulating the protein expression of α-SMA and Runx2.

Effect of Jianpi Xiaoai Prescription on Activation of HFL1 into CAFs Induced by Exosomes in Colon Cancer / 中国实验方剂学杂志

Objective:To observe the effect of Jianpi Xiaoai prescription on the activation of normal human embryonic lung fibroblasts （HFL1） into tumor-associated fibroblasts （CAFs） induced by human colon cancer cells （HCT116） derived exosomes. Method:SD rats were gavaged with 13.1 g·kg-1 of Jianpi Xiaoai prescription to prepare drug-containing serum， and HCT116 cell exosomes-containing 10% exosomes-free serum and 20% Jianpi Xiaoai prescription drug serum were isolated by ultra-high speed centrifugation. The particle size distribution of exosomes were detected by Nanoparticle tracking analyzer （Zetaview）， and the exosomes' marker proteins apoptotic transfer gene 2 interaction protein X （Alix）， heat shock protein 70 （HSP70）， and tumor-susceptibility gene 101 （TSG101） were identified by Western blot， and the uptake of exosomes labeled with cell membrane staining kit （PKH67） by HFL1 was observed by fluorescence microscope. HFL1 cells were divided into six groups： the blank group， the transforming growth factor-β1 （TGF-β1） group， the TGF-β1 combined with HCT116 exosomes of 2 mg·L-1 group， the TGF-β1 combined with HCT116 exosomes of 4 mg·L-1 group， the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 2 mg·L-1 group， and the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 4 mg·L-1 group， and all groups were cultivated for 48 h. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to determine the protein and mRNA expressions of α-smooth muscle actin （α-SMA）. Result:The particle size distribution detected by Zetaview was mainly between 50-100 nm， and the exosomes were verified based on the expressions of marker proteins Alix， HSP70 and TSG101. After co-incubation of HFL1 cells with exosomes， a large number of exosomes were absorbed by HFL1 cells under fluorescence microscope. Compared with the blank control group， the protein and mRNA expressions of α-SMA in the TGF-β1 group and TGF-β1 combined with HCT116 exosome groups were increased （P<0.01）. Compared with the TGF-β1 combined with HCT116 exosome groups， the protein and mRNA expressions of α-SMA were decreased in the TGF-β1 combined with Jianpi Xiaoai prescription exosome groups （P<0.01）. Conclusion:Human colon cancer cell exosomes combined with TGF-β1 can induce the activation of HFL1 into CAFs， and Jianpi Xiaoai prescription can reduce the activation of HFL1 by affecting the expressions of α-SMA， thus antagonizing the lung metastasis of colon cancer.

Expression and significance of hypoxia inducible factor-1α and α-smooth-muscle actin in cervical cancer / 中国医师进修杂志

Objective To examine the expression levels of hypoxia inducible factor (HIF)-1α and α-smooth-muscle actin (SMA) in both cervical cancer tissues with concurrent chemoradiotherapy and non-cervical cancer tissues,and assess the clinical significance in cervical cancer.Methods The immune-histochemistry was used to detect HIF-1α and α-SMA in 68 cases of cervical cancer tissues and 56 cases of non-cervical cancer tissues (including normal cervical tissues,hysteromyoma and cervical intraepithelial neoplasias) from January 2013 to January 2014 in the First Affiliated Hospital of Hebei Northern University.Results The positive expression rates of HIF-1α and α-SMA in cervical cancer tissues and non-cervical cancer tissues were 58.8％ (40/68),39.3％ (22/56) and 54.4％ (37/68),35.7％ (20/56) respectively;the differences were significant statistically (P＜ 0.05).The expression of HIF-1α was positively correlated with the expression of α-SMA protein in cervical cancer tissues (r =0.376,P =0.001).The positive expression of HIF-1α was closely related to International Federation of Gynecology and Obstetrics (FIGO) stages (r =0.371,P =0.004),lymph node metastasis (r =0.243,P =0.039),abdominal aortic parathyroid lymph node metastasis (r =0.286,P =0.014) and serum squamous cell carcinoma antigen (SCC-Ag) (r =0.271,P =0.020),but it was not correlated with age and tumor size (P ＞ 0.05).The positive expression of α-SMA was associated with lymph node metastasis (r =0.363,P =0.001),abdominal aortic parathyroid lymph node metastasis (r =0.271,P =0.020) and SCC-Ag (r =0.272,P =0.020),but it was not correlated with age,FIGO stage and tumor size (P ＞ 0.05).The shortterm effect rates of concurrent chemoradiotherapy in cervical cancer tissues with positive and negative expression of HIF-1α and α-SMA were 27.5％ (11/40),21.6％ (8/37),and 64.3％ (18/28) and 48.4％ (15/31),and there were statistical differences (P ＜ 0.05).The medial follow-up time of 68 cases was 60 months,and the 5-year survival rate was 52.9％ (36/68).The 5-year survival rates of the patients with positive expression of HIF-1α and α-SMA were 42.5％ (17/40) and 40.5％ (15/31),the 5-year survival rates of the patients with negative expression of HIF-1α and α-SMA were 67.9％ (19/28) and 67.7％ (21/31),and the differences were statistically significant separately (x2 =4.091 and 4.573,P =0.043 and 0.032).Conclusions The elevation of HIF-1α and α-SMA may be used to predict the development and prognosis of cervical cancer with concurrent chemoradiotherapy.

Expression and significance of hypoxia inducible factor-1α and α-smooth-muscle actin in cervical cancer / 中国医师进修杂志

Objective@#To examine the expression levels of hypoxia inducible factor (HIF)-1α and α-smooth-muscle actin (SMA) in both cervical cancer tissues with concurrent chemoradiotherapy and non-cervical cancer tissues, and assess the clinical significance in cervical cancer.@*Methods@#The immune-histochemistry was used to detect HIF-1α and α-SMA in 68 cases of cervical cancer tissues and 56 cases of non-cervical cancer tissues (including normal cervical tissues, hysteromyoma and cervical intraepithelial neoplasias) from January 2013 to January 2014 in the First Affiliated Hospital of Hebei Northern University.@*Results@#The positive expression rates of HIF-1α and α-SMA in cervical cancer tissues and non-cervical cancer tissues were 58.8% (40/68), 39.3% (22/56) and 54.4% (37/68), 35.7% (20/56) respectively; the differences were significant statistically (P<0.05). The expression of HIF-1α was positively correlated with the expression of α-SMA protein in cervical cancer tissues (r = 0.376, P =0.001). The positive expression of HIF-1α was closely related to International Federation of Gynecology and Obstetrics (FIGO) stages (r = 0.371, P = 0.004), lymph node metastasis (r = 0.243, P = 0.039), abdominal aortic parathyroid lymph node metastasis (r = 0.286, P = 0.014) and serum squamous cell carcinoma antigen (SCC-Ag) (r = 0.271, P = 0.020), but it was not correlated with age and tumor size (P>0.05). The positive expression of α-SMA was associated with lymph node metastasis (r = 0.363, P = 0.001), abdominal aortic parathyroid lymph node metastasis (r = 0.271, P = 0.020) and SCC-Ag (r = 0.272, P = 0.020), but it was not correlated with age, FIGO stage and tumor size (P>0.05) . The short-term effect rates of concurrent chemoradiotherapy in cervical cancer tissues with positive and negative expression of HIF-1α and α-SMA were 27.5% (11/40), 21.6% (8/37), and 64.3% (18/28) and 48.4% (15/31), and there were statistical differences (P<0.05). The medial follow-up time of 68 cases was 60 months, and the 5-year survival rate was 52.9% (36/68). The 5-year survival rates of the patients with positive expression of HIF-1α and α-SMA were 42.5% (17/40) and 40.5% (15/31), the 5-year survival rates of the patients with negative expression of HIF-1α and α-SMA were 67.9% (19/28) and 67.7% (21/31), and the differences were statistically significant separately (χ2 = 4.091 and 4.573, P = 0.043 and 0.032).@*Conclusions@#The elevation of HIF-1α and α-SMA may be used to predict the development and prognosis of cervical cancer with concurrent chemoradiotherapy.

Effects of moderate treadmill exercise on alpha-smooth muscle actin and type iv collagen in the liver of type 2 diabetic rats / 中国组织工程研究

BACKGROUND: Diabetes-induced liver damage is easy to be ignored in the early stage. Exercise therapy can increase the sensitivity of insulin, which is an important means of prevention and treatment of diabetes. OBJECTIVE: To explore the effect of moderate intensity exercise intervention on liver injury during the occurrence of diabetes mellitus. METHODS: The experimental protocol was approved by the Laboratory Animal Care Ethics Committee of Wuhan Sports University. Thirty SPF Sprague-Dawley rats were divided into three groups: normal control group, diabetic control group and diabetic exercise intervention group. Normal control group was fed with normal diet, with no exercise. Diabetes control group was fed with high sugar and high fat diet for 8 weeks, followed by a small dose of streptozotocin (30 mg/kg) injected intraperitoneally, with o exercise. Diabetes exercise intervention group was fed and injected in the same way as diabetes control group, and at the same time carried out moderate intensity treadmill training. After 7 days of streptozotocin injection, hematoxylin-eosin and Masson staining were used to observe the cell morphology, the expression of α-smooth muscle actin was observed by immunohistochemistry, and orbital blood samples were collected to detect the concentration of serum type IV collagen using ELISA. RESULTS AND CONCLUSION: Compared with the normal control group, hematoxylin-eosin staining showed that the structure of liver lobule in the diabetic control group was disordered, a large number of fat vacuoles were seen, and the bile duct in the portal area was obviously proliferated; Masson staining that there were showed fat vacuoles, the structure of liver lobule was seriously damaged, and blue stained collagen fibers were seen in the portal area, and light blue stained collagen fibers were seen between liver cells. The above pathological changes were alleviated in the diabetic exercise intervention group, and the fatty degeneration of liver cells was obviously reduced. The expression of α-smooth muscle actin in the diabetic control group and diabetic exercise intervention group was significantly higher than that in the normal control group (P < 0.05), and that in the diabetic exercise intervention group was significantly lower than that in the diabetic control group (P < 0.05). The level of type IV collagen in the diabetic exercise intervention group was significantly lower than that in the diabetic control group (P < 0.05), to slow down the progress of fibrosis. To conclude, moderate intensity exercise has a good effect on streptozotocin induced liver fibrosis in diabetic rats.

Therapeutic effect of human adipose stem cells derived exosomes on carbon tetrachloride induced liver fibrosis in rats / 中国组织工程研究

BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets. OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.