Objective To investigate pathological changes of lens epithelium cells (LECs) and expressions of matrix metalloproteinase-3 (MMP-3),alpha-smooth muscle actin (α-SMA),and fibronectin (Fn) in lens epithelial cells of experimental diabetic cataract rats,and to evaluate the roles of MMP-3,α-SMA,and Fn in the pathogenesis of diabetic cataract.Methods A total of 105 healthy Sprague-Dawley (SD) rats without lens diseases was randomly divided into normal control (n =45) and diabetic model (n =60) groups.Diabetic model rats were subjected to intraperitoneal injection of streptozotocin (STZ) (60 mg/kg),and those in the normal control group received injection with 0.1 mmol/L citric acid buffer solution of the same volume.The diabetic models were affirmed upon a fasting blood ≥ 16.65 mmol/L at the 3rd days after the injection.Once a week,the changes of blood glucose and body weight were monitored and the progression of cataract formation in both lenses of all rats was recorded with slit lamp observation.At end of 4 weeks,8 weeks,and 12 weeks after STZ injection,lenses were isolated and embedded in paraffin.The LECs histopathology was examined with HE staining.Immunohistochemical method was used to detect expressions of MMP-3,α-SMA,and Fn in normal and diabetic LECs.The related comparisons and statistic analysis were carried out.Results The lenses of control group were always completely transparent throughout the period of experiment with 31.48％,77.78％,and 100％ of lenses in diabetic model group swelling at 4th,8th,and 12th week,respectively.Under the light microscopic level,it has been showed that lens epithelium cells,which was occurred aggregate plaque and arranged in many layers,presented some morphologic characteristics of fibroblasts by HE staining.In control group LECs regularly,MMP-3,α-SMA,and Fn did not express equally;MMP-3,α-SMA,and Fn expressions increased obviously highly,difference had statistical significance compared to diabetic cataract group LECs (P ＜ 0.01).With the development of course of disease,the differences in expression of MMP-3,α-SMA,and Fn were significant between the diabetic cataract LECs and the control group (P ＜ 0.01).During the progression of diabetic cataract,the expression of Fn was positively correlated with that of α-SMA and MMP-3 (r =0.994,P ＜ 0.01;r =0.993,P ＜ 0.01).Conclusions The diabetic cataract liyingbody animal model was established successfully,which has laid down necessary basis to expound the morbidity mechanism of diabetic cataract.In the lens of diabetic cataract group,the expressions of α-SMA,MMP-3,and Fn were significantly increased.They participated in the occurrence and development of diabetic cataract.The expression of Fn was positively correlated with α-SMA and MMP-3.The lens epithelium cells which took on elongated aspect like fibroblast cells appeared epithelial-mesenchymal transformation (EMT),which mediated the occurrence and development of LECs fibrosis.

BACKGROUND:Some studies have indicated that different genes in tooth germ tissue play a role at different time, contributing to tooth germ development.
OBJECTIVE:To observe the expressions of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 at different stage of in vitro culture of tooth germ cells.
METHODS:RNA from tooth germ cells was extracted at days 1, 3, 6 of in vitro culture. After reverse transcription, real-time quantitative PCR detection was adopted to measure relative expression of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 mRNA.
RESULTS AND CONCLUSION:Dentin matrix protein 1, enamel protein, and col agen I mRNA expressions increased with culture time, and reached the peak at day 3 (P<0.05), whereas homeobox gene 1 mRNA decreased with culture time (P<0.05).

BACKGROUND:With the promotion of 3D printing technology, 3D printing scaffolds for bone tissue engineering have become the new ideas for jaw bone repair. OBJECTIVE:To compare the physical and biological properties of sheep vertebral bone meal/polyvinyl alcohol (PVA) scaffold, nano-hydroxyapatite (nHA)/PVA scaffold, and sheep vertebral bone meal/PVA nonporous bone plate. METHODS:3D printing technology was used to print sheep vertebral bone meal/PVA scaffold, nHA/PVA scaffold, and sheep vertebral bone meal/PVA nonporous bone plate. Porosity, morphology, water absorption rate and mechanical properties of different scaffolds were detected. Three kinds of scaffolds were al used to culture bone marrow mesenchymal stem cel s, and cel proliferation ability was detected using cel counting kit-8 at 1, 4, 7 days of culture. RESULTS AND CONCLUSION:Under scanning electron microscope, the sheep vertebral bone meal/PVA scaffold and nHA/PVA scaffold exhibited regular and interconnected pores with good continuity and clear network structure;the sheep vertebral bone meal/PVA nonporous bone plate had no obvious pores;however, it had dense and evenly distributed micropores with different sizes on its surface. The porosity of nHA/PVA scaffold was lower than that of the sheep vertebral bone meal/PVA scaffold (P<0.05). The water absorption rate was highest for the nHA/PVA scaffold fol owed by the sheep vertebral bone meal/PVA scaffold and the sheep vertebral bone meal/PVA nonporous bone plate (P<0.05). In contrast, the scaffold toughness was highest for the sheep vertebral bone meal/PVA nonporous bone plate, fol owed by the sheep vertebral bone meal/PVA scaffold and nHA/PVA scaffold. In addition, the cel proliferation activity of cel s cultured on the sheep vertebral bone meal/PVA scaffold was significantly higher than that cultured on the other two kinds of scaffolds. Taken together, the 3D printing sheep vertebral bone/PVA scaffold has good physical and chemical performance.

METHODS:Lentiviral vectors carrying bFGF and BMP-2 were constructed to transfect sheep bone marrow mesenchymal stem cells. cells were divided into four groups:bFGF group, BMP-2 group, co-transfection group BACKGROUND:Basic fibroblast growth factor (bFGF) can promote the proliferation of bone marrow stromal cells, and bone morphogenetic protein-2 (BMP-2) has an important significance in the induction of new bone formation.
OBJECTIVE:To analyze the effects of bFGF, BMP-2 and their co-transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to compare the relative expressions of col agen I, osteocalcin and osteopontin before and after celltransfection, thereby providing theoretical implications for seed cells in the construction of tissue-engineered bone.
and control group. The RNA was extracted using real-time quantitative PCR to detect mRNA levels of col agen I, osteocalcin, and osteopontin.
RESULTS AND CONCLUSION:Significant difference in non-specific osteogenic gene expressions was found among the four groups (P<0.05). bFGF and BMP-2 showed an interaction (P<0.05). Expressions of col agen I and osteocalcin in the co-tranfection group were higher than those in the other three groups (P<0.05), but osteopontin expression exhibited no difference (P>0.05). In vitro experiments showed that the relative expression of col agen I, osteocalcin and osteopontin were higher in the co-transfection group, indicating the cells from the co-transfection group have strongest osteogenic capacity that are suitable for seed cells for bone tissue engineering.

OBJECTIVE To investigate the relationship between the skin prick test and clinical characteristics in patients with chronic sinusitis with nasal polyps(CRSwNP).METHODS Sixty cases with CRSwNP and 40 control samples underwent skin prick test with standardized allergens provided by AllergoPhamar Company,the results and their clinical characteristics are analyzed.RESULTS The positive rate of skin prick test was 81.7% and 17.5% in CRSwNP group and control group respectively(?2 =40.104,?

OBJECTIVE: To study the expression of insulin-like growth factor-1 receptor(IGF-1R) in nasal polyps and its relationship with allergic rhinitis. METHOD: The mRNA and protein expression of IGF-1R in 40 cases (20 cases with allergic rhinitis and 20 cases without) nasal polyps tissue (the CRSWNP group) and 20 middle turbinate tissue samples (the control group) were examined by fluorescence quantitative polymerase chain reaction (FQ-PCR) and immunohistochemistry. RESULT: The positive staining rate of IGF-1R protein of nasal polyps tissue is 70.8% and that of control is 12.3%, the expression of IGF-1R mRNA of nasal polyp is 0.748 +/- 0.111,which is significant higher than that of the control group is 0.107 +/- 0.208 (P < 0.01). No significant difference of the expression of IGF-1R mRNA between with and without allergic rhinitis cases and between with and without endoscopy sinus surgery history cases in the CRSWNP group. CONCLUSION The overexpression of IGF-1R maybe play important roles in the formation of nasal polyp. Hypersensitivity reaction type I mediated by IgE has no contribution to the overexpression of the IGF-1R.
Adult ; Case-Control Studies ; Female ; Humans ; Hypersensitivity ; pathology ; Male ; Middle Aged ; Nasal Polyps ; immunology ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptor, IGF Type 1 ; metabolism ; Young Adult

Objective: o study the effect of cleaning treatment with hydrofluoric acid (HF) on the surface and bonding strength of IPS e.max and Vita Mark II ceramic inlays. Methods: Fifty pieces of IPS e.max and Vita Mark II ceramic inlay specimens were made separately using CAD/CAM. After uniformly bonding surfaces using 9% HF etching, they were randomly divided into an untreated control group (group A) and the following experimental groups: neutralizing powder (B group), 37% phosphoric acid (group C), ultrasonic cleaning (group D) and neutralizing powder + 37% phosphoric acid + ultrasonic cleaning (group E). Each set of 8 specimens was bonded to Variolink N resin adhesive under standard conditions. The shear adhesive strength was measured after exposure to a constant-temperature water bath at 37 ℃ for 24 h. The location of the fracture and the type of adhesion failure were recorded. The shear adhesion and the average strength of the connection were analyzed. The remaining 2 specimens were used for scanning electron microscopy (SEM) to observe the surface morphology, including the crystal structure, pore pattern, and residue. Results : The results were similar for the IPS e.max and Vita Mark II inlays. The maximum bond strength was observed in the IPS e.max ceramic inlays in group E, with an average bond strength 11.96 MPa higher than that in group A. Among the Vita Mark II porcelain inlays, the maximum bond strength was observed in group E. The average bond strength was 9.74 MPa higher than that in group A. The results of the statistical analysis were similar for the IPS e.max and Vita Mark II porcelain inlays, with significant differences in the bond strengths between groups C, D, and E and the control group (P < 0.05). There was no significant difference in the adhesive strength between groups B and A. At the same time, there was no significant difference in the bond strength between the treatment groups B, C, D, and E (P > 0.05). SEM revealed that the pores on the surface of ceramics subjected to the acid etching treatment were broadened and uniform, with less residue than observed in the control group. The effects of treatments D and E were the best. Conclusion The HF etching treatment can enhance the bonding strength of IPS e.max and Vita Mark Ⅱ ceramic inlays while leaving little residue, and the joint strength is highest when the joints are treated together.

Objective: To investigate the effects of over expression and low expression of antisense transcripts of circular RNA cerebellar degeneration associated protein 1 (CDR1as) in Balb/C mouse bone marrow mesenchymal stem cells (BMSCs) on factors related to osteogenesis and angiogenesis. Methods: BMSCs were cultured and identified in vitro. The lentiviral (LV) vector containing the overexpressed and silenced circRNA CDR1as genes and the control lentivirus were respectively transfected into mouse BMSCs, and stable cell lines were screened. The cells were divided into the circRNACDR1as over expression group and the over expression control group, and the CircRNACDR1as low expression group and the low expression control group. The components were stained with Alizarin Red S and alkaline phosphatase after 14 and 21 days of osteoinduction; qRT-PCR was used to detect the target genes circRNA CDR1as, osteogenic differentiation markers alkaline phosphatase (ALP), runt- related transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN), osterix(Osx), collagen I (COL-1), and the mRNA expression levels of vascular endothelial grown factor (VEGF) and angiogenin-1 (Ang-1). Results: The results of alizarin red staining and alkaline phosphatase staining showed that the extracellular matrix calcium precipitation and ALP staining area of the over expression experimental group was greater than its control group, and those of the low expression experimental group was less than its control group. As the number of days of osteogenic induction increased, the calcium precipitation and ALP staining in each group also increased. RT-PCR results showed that the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in the over expression experimental group BMSCs were significantly increased (P<0.001). In the low expression experimental group, the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in BMSCs were significantly reduced (P<0.001). Conclusion Over expression of the circRNA CDR1as gene promotes the osteogenic differentiation and angiogenesis of BMSCs. Low expression of the circRNA CDR1as gene inhibits the osteogenic differentiation and angiogenesis of BMSCs.

BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function. OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased(P<0.05)in the miR-378a transfection group.(4)It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.

Objective: To investigate the level of hypoxia inducible factor-1α (HIF-1α) on osteoblasts and angiogenesis-associated cytokines in bone marrow mesenchymal stem cells (BMSCs) from SD rats. Methods: BMSCs were isolated and cultured and identified by flow cytometry. Plasmid vectors containing upregulated and downregulated HIF-1α gene and a control vector were constructed. The plasmids were transfected into BMSCs by Lipofectamine®LTX transfection reagent, and the cells were divided into an overexpression experimental group, an overexpression control group, a low expression experimental group and a low expression control group. All components were stained with a lizarin red 3 d and 7 d after osteogenesis induction. The mRNA expression levels of the target gene HIF-1α, osteogenic differentiation-specific markers, including Runt-related transcription factor 2 (Runx2) and angiogenic markers, including platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β (TGF-β), were detected by RT-PCR. Western blot was used to detect the protein expression of the target proteins HIF-1α, Runx2, and PDGF-BB. Results: The CD29- and CD45-positivity rates of BMSC surface markers identified by flow cytometry were 98.2% and 4.2%, respectively. RT-PCR results showed that the mRNA expression of HIF-1α, Runx2, TGF-β and PDGF-BB was observably increased (P < 0.001). The mRNA expression levels of HIF-1α, Runx2, TGF-β and PDGF-BB in BMSCs from the low expression experimental group were significantly reduced (P < 0.001). Western blot results showed that the expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the overexpression experimental group were all increased (P < 0.001). The expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the low expression experimental group were reduced (P < 0.001). Alizarin red staining results showed that the area of calcium nodules in the low expression experimental group was smaller than that in low expression control group, the area of red calcium nodules in the over expression experimental group was larger than that in over expression control group, and with the increase of osteogenic induction time, the calcification area of each group also increased. Conclusion Upregulation and downregulation of HIF-1α can regulate the osteogenic differentiation and the expression of angiogenesis related factors of BMSCs.