MiR-149-3p promotes the cisplatin resistance and EMT in ovarian cancer through downregulating TIMP2 and CDKN1A.

BACKGROUND: Ovarian cancer (OC), a kind of gynecological cancer, is characterized by high mortality rate, with microRNAs (miRNAs) playing essential roles in it. However, the clinical significance of miRNAs and their molecular mechanisms in OC are mostly unknown. METHODS: miR-149-3p expression was predicted through Gene Expression Omnibus (GEO) data in OC and confirmed by q-PCR in various OC cells and tissues from patients with different clinical characteristics. Moreover, its roles in terms of proliferation, migration and invasion were measured by CCK-8, colony formation, wound healing and transwell assays in OC cells including cisplatin-resistant and cisplatin-sensitive cells. And its effect on epithelial-mesenchymal transition was also assessed through detecting related protein expression. Additionally, its potential targets were verified by dual luciferase assay and Ago-RIP assay. Finally, its oncogenic functions were explored in vivo. RESULTS: In data from GSE79943, GSE131790, and TCGA, miR-149-3p was found to be highly expressed in OC tissues and associated with poor survival. In metastasis and chemoresistant tissues and cisplatin-resistant OC cells, its high expression was confirmed. In terms of tumorigenic effects, miR-149-3p knockdown in cisplatin-resistant OC cells inhibited its cisplatin resistance and other malignant phenotypes, while miR-149-3p overexpression in cisplatin-resistant OC cells led to contrary results. Mechanistically, miR-149-3p targeted 3'UTR of CDKN1A and TIMP2 to function as an oncogenic miRNA. CONCLUSION: In brief, miR-149-3p promoted cisplatin resistance and EMT in OC by downregulating CDKN1A and TIMP2, which might provide a potential therapeutic target for OC treatment.

Arginine and lysine methylation of MRPS23 promotes breast cancer metastasis through regulating OXPHOS.

Mitochondrial oxidative phosphorylation (OXPHOS) is a vital regulator of tumor metastasis. However, the mechanisms governing OXPHOS to facilitate tumor metastasis remain unclear. In this study, we discovered that arginine 21(R21) and lysine 108 (K108) of mitochondrial ribosomal protein S23 (MRPS23) was methylated by the protein arginine methyltransferase 7 (PRMT7) and SET-domain-containing protein 6 (SETD6), respectively. R21 methylation accelerated the poly-ubiquitin-dependent degradation of MRPS23 to a low level. The MRPS23 degradation inhibited OXPHOS with elevated mtROS level, which consequently increased breast cancer cell invasion and metastasis. In contrast, K108 methylation increased MRPS23 stability, and K108 methylation coordinated with R21 methylation to maintain a low level of MRPS23, which was in favor of supporting breast cancer cell survival through regulating OXPHOS. Consistently, R21 and K108 methylation was correlated with malignant breast carcinoma. Significantly, our findings unveil a unique mechanism of controlling OXPHOS by arginine and lysine methylation and point to the impact of the PRMT7-SETD6-MRPS23 axis during breast cancer metastasis.

Groundwater Quality in Agricultural Lands Near a Rapidly Urbanized Area, South China.

Understanding the groundwater quality and its factors is a key issue in the context of the use and protection of groundwater resources in agricultural areas near urbanized areas. This study assessed the groundwater quality in agricultural areas in the Pearl River Delta (PRD) by a fuzzy synthetic evaluation method and determined the main factors controlling the groundwater quality by principal component analysis (PCA). Results showed that approximately 85% of groundwater sites in agricultural lands in the PRD were good-quality (drinkable). Drinkable groundwater was 95% and 80% in fissured aquifers and porous aquifers, respectively. Poor-quality groundwater in porous aquifers was controlled by four factors according to the PCA, including the seawater intrusion; the lateral recharge and irrigation of surface water and geogenic sources for As, Fe, NH4+, and Mn; the wastewater infiltration; and the geogenic sources for iodide. By contrast, another four factors, including the infiltration of wastewater and agricultural fertilizers, the geogenic sources for heavy metals, the geogenic sources for iodide, and the irrigation of contaminated river water, were responsible for the poor-quality groundwater in fissured aquifers. Therefore, in the future, the groundwater protection in agricultural lands in the PRD should be strengthened because the majority of groundwater in these areas was good-quality and suitable for drinking and agricultural purposes. In addition, poor-quality groundwater in agricultural lands in the PRD was a small proportion and negligible because the factors for poor-quality groundwater are complicated.

The roles of long noncoding RNAs in breast cancer metastasis.

Breast cancer is the most significant threat to female health. Breast cancer metastasis is the major cause of mortality in breast cancer patients. To fully unravel the molecular mechanisms that underlie the breast cancer cell metastasis is critical for developing strategies to improve survival and prognosis in breast cancer patients. Recent studies have revealed that the long noncoding RNAs (lncRNAs) are involved in breast cancer metastasis through a variety of molecule mechanisms, though the precise functional details of these lncRNAs are yet to be clarified. In the present review, we focus on the functions of lncRNAs in breast cancer invasion and metastasis, with particular emphasis on the functional properties, the regulatory factors, the therapeutic promise, as well as the future challenges in studying these lncRNA.

Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

Arginine methyltransferase PRMT7 is associated with human breast cancer metastasis. Endosomal FAK signalling is critical for cancer cell migration. Here we identified the pivotal roles of PRMT7 in promoting endosomal FAK signalling activation during breast cancer metastasis. PRMT7 exerted its functions through binding to scaffold protein SHANK2 and catalyzing di-methylation of SHANK2 at R240. SHANK2 R240 methylation exposed ANK domain by disrupting its SPN-ANK domain blockade, promoting in co-accumulation of dynamin2, talin, FAK, cortactin with SHANK2 on endosomes. In addition, SHANK2 R240 methylation activated endosomal FAK/cortactin signals in vitro and in vivo. Consistently, all the levels of PRMT7, methylated SHANK2, FAK Y397 phosphorylation and cortactin Y421 phosphorylation were correlated with aggressive clinical breast cancer tissues. These findings characterize the PRMT7-dependent SHANK2 methylation as a key player in mediating endosomal FAK signals activation, also point to the value of SHANK2 R240 methylation as a target for breast cancer metastasis.

Arginine hypomethylation-mediated proteasomal degradation of histone H4-an early biomarker of cellular senescence.

Senescence is accompanied with histones level alteration; however, the roles and the mechanisms of histone reduction in cellular senescence are largely unknown. Protein arginine methyltransferase 1 (PRMT1) is the major enzyme that generates monomethyl and asymmetrical dimethyl arginine. Here we showed that abrogation of PRMT1-mediated senescence was accompanied with decreasing histone H4 level. Consistently, under multiple classic senescence models, H4 decreasing was also been found prior to the other 3 core histones. Noticeably, asymmetric demethylation of histone H4 at arginine 3 (H4R3me2as), catalyzed by PRMT1, was decreased prior to histone H4. In addition, we showed that the PRMT1-mediated H4R3me2as maintained H4 stability. Reduction of H4R3me2as level increased the interaction between proteasome activator PA200 and histone H4, which catalyzes the poly-ubiquitin-independent degradation of H4. Moreover, H4 degradation promoted nucleosome decomposition, resulting in increased senescence-associated genes transcription. Significantly, H4 was restored by 3 well-informed anti-aging drugs (metformin, rapamycin, and resveratrol) much earlier than other senescence markers detected under H2O2-induced senescence. Thus, we uncovered a novel function of H4R3me2as in modulation of cellular senescence via regulating H4 stability. This finding also points to the value of histone H4 as a senescence indicator and a potential anti-aging drug screening marker.

Arginine methylation-dependent LSD1 stability promotes invasion and metastasis of breast cancer.

Histone lysine demethylase 1 (LSD1), the first identified histone demethylase, is overexpressed in multiple tumor types, including breast cancer. However, the mechanisms that cause LSD1 dysregulation in breast cancer remain largely unclear. Here, we report that protein arginine methyltransferase 4 (PRMT4 or CARM1) dimethylates LSD1 at R838, which promotes the binding of the deubiquitinase USP7, resulting in the deubiquitination and stabilization of LSD1. Moreover, CARM1- and USP7-dependent LSD1 stabilization plays a key role in repressing E-cadherin and activating vimentin transcription through promoter H3K4me2 and H3K9me2 demethylation, respectively, which promotes invasion and metastasis of breast cancer cells. Consistently, LSD1 arginine methylation levels correlate with tumor grade in human malignant breast carcinoma samples. Our findings unveil a unique mechanism controlling LSD1 stability by arginine methylation, also highlighting the role of the CARM1-USP7-LSD1 axis in breast cancer progression.

Clinical study on the posterior occlusal plane and morphology of temporomandibular joint / Estudio clínico sobre el plano oclusal posterior y la morfología de la articulación temporomandibular

The objective of the study was to investigate the morphological features of the temporomandibular joint in adult patients with posterior occlusal plane and different inclinations. Fifty five skeletal I occlusion patients with average were included, shooting CBCT in the intercuspal position, divided into 3 groups according to OPP-FH angle, and measurement of the temporomandibular joint morphology was calculated with cone beam computed tomography (CBCT) special software Invivo 5.0, Statistical analysis of three groups of data using SPSS23.0. The condyle usually locates in the middle of the fossa, the maximum axial area of the condyle (A) was statistically significant between the group 1 and the group 3. The internal and external diameter (MD) of the condyle was statistically significant between group 1 and group 3, and group 2 and group 3. The bilateral TMJ morphological features of the three groups were basically symmetrical. The position of the condyle in the fossa is mostly centered, and some of the posterior, the maximum axial area and the internal and external diameter of the condyle are different in three groups.

El objetivo de este estudio fue investigar las características morfológicas de la articulación temporomandibular (ATM) en pacientes adultos con plano oclusal posterior y diferentes inclinaciones. Se incluyeron 55 pacientes con oclusión esquelética tipo I, visualizados por tomografía computarizada de haz cónico (CBCT) en posición intercuspiana, y se dividieron en 3 grupos según el ángulo OPP-FH. La medición morfológica de la articulación temporomandibular se calculó con CBCT y mediante el software especial Invivo 5.0. El análisis estadístico de datos se realizó con el software SPSS 23.0. El cóndilo de la mandíbula generalmente se ubica en el centro de la fosa; el área axial máxima del cóndilo de la mandíbula (A) fue estadísticamente significativa entre los grupos 1 y 3. Los datos de los diámetros medial y lateral (DM) del cóndilo de la mandíbula fueron estadísticamente significativos entre los grupos 1 y 3 y los grupos 2 y 3. Las características morfológicas de la ATM de los tres grupos fueron básicamente simétricas. La posición del cóndilo de la mandíbula en la fosa fue principalmente centrada, y parte del área axial máxima, posterior y los diámetros medial y lateral del cóndilo de la mandíbula fueron diferentes en los tres grupos.

Expression of Ihh signaling pathway in condylar cartilage after bite-raising in adult rats.

Temporomandibular joint osteoarthritis (TMJOA) is a complex inflammatory condition with multiple factors and degenerative processes co-occurring. However, its pathogenesis remains uncertain. The purpose of the study was to observe the expression of Indian hedgehog (Ihh) signal related molecules in TMJOA induced by bite-raising and to study the effect and mechanism of Ihh signaling. Our research indicated that Ihh signaling pathway can be activated in condylar cartilage induced by bite-raising. The histological analysis showed TMJOA-like structural changes of condylar cartilage in experiment groups. Ihh, Smoothened (Smo), and Gli zinc finger transcription factors-1 (Gli-1) were activated in the experimental groups, and the expression levels increased significantly over time, whereas the sham control groups showed no fluctuation. Additionally, the expression levels of matrix metalloproteinase-13 (MMP-13) and cysteinyl aspartate specific proteinase-3 (Caspase-3) in the experiment groups increased in a time-dependent manner compared with the matched sham control groups. In conclusion, our results indicated that the Ihh signaling pathway may activate the occurrence of TMJOA by mediating the hypertrophy of chondrocytes, which may be an important regulatory mechanism and potential therapeutic target in the repair of condylar cartilage.

LncRNA ANCR down-regulation promotes TGF-ß-induced EMT and metastasis in breast cancer.

Epithelial to mesenchymal transition (EMT) is a progression of cellular plasticity critical for development, differentiation, cancer cells migration and tumor metastasis. As a well-studied factor, TGF-ß participates in EMT and involves in physiological and pathological functions of tumor progression. Accumulating evidence indicates that long noncoding RNAs(lncRNAs) play crucial roles in EMT and tumor metastasis. Here, we find that lncRNA ANCR participates in TGF-ß1-induced EMT. By our ChIP and Real-time PCR assays, we reveal that TGF-ß1 down-regulates ANCR expression by increasing HDAC3 enrichment at ANCR promoter region, which decreases both H3 and H4 acetylation of ANCR promoter. In addition, by western blot and transwell assays, we indicate that ectopic expression of ANCR partly attenuates the TGF-ß1-induced EMT. Downstream, ANCR inhibits breast cancer cell migration and breast cancer metastasis by decreasing RUNX2 expression in vitro and in vivo. Thus, our study identifies ANCR, as a new TGF-ß downstream molecular, is essential for TGF-ß1-induced EMT by decreasing RUNX2 expression. These results implicate that ANCR might become a prognostic biomarker and an anti-metastasis therapy target for breast cancer.

GSK3ß activity is essential for senescence-associated heterochromatin foci (SAHF) formation induced by HMGA2 in WI38 cells.

Cellular senescence is an irreversible form of cell cycle arrest, which is often characterized by domains of facultative heterochromatin substructures also known as senescence-associated heterochromatin foci (SAHF). SAHF assembly is likely mediated through the downregulation of the Wnt pathway, which inhibits Glycogen Synthase Kinase 3 Beta (GSK3ß) in cells undergoing replicative senescence. Alternatively, High Mobility Group AT-Hook 2 (HMGA2) can also induce SAHF formation in primary cells, through a process in which the involved cell signaling pathway is unknown. Accordingly, it is important to determine whether GSK3ß and the Wnt pathway are necessary during HMGA2-induced SAHF formation. In this study, we developed a senescence model for SAHF assembly in WI38 cell through ectopic expression of HMGA2. In this model, typical senescent features were identified, including elevated SA-ß-galactosidase staining and the downregulation of the Wnt pathway. We also showed that the GSK3ß inhibitor LiCl can partly disable SAHF formation through the HMGA2 overexpression in WI38 cells. However, the disabled SAHF formation resulting from the inactivity of GSK3ß in our senescence model cannot be restored through ectopic overexpression of Catenin Beta 1 (CTNNB1), a downstream transcription factor of the Wnt pathway. This indicates that the GSK3ß activity alone, and not those of downstream target genes, is crucial for the HMGA2-induced SAHF formation following the downregulation of the Wnt pathway.

Methylation of arginine by PRMT1 regulates Nrf2 transcriptional activity during the antioxidative response.

The cap 'n' collar (CNC) family of transcription factors play important roles in resistance of oxidative and electrophilic stresses. Among the CNC family members, NF-E2-related factor 2 (Nrf2) is critical for regulating the antioxidant and phase II enzymes through antioxidant response element (ARE)-mediated transactivation. The activity of Nrf2 is controlled by a variety of post-translational modifications, including phosphorylation, ubiquitination, acetylation and sumoylation. Here we demonstrate that the arginine methyltransferase-1 (PRMT1) methylates Nrf2 protein at a single residue of arginine 437, both in vitro and in vivo. Using the heme oxygenase-1 (HO-1) as a model of phase II enzyme gene, we found that methylation of Nrf2 by PRMT1 led to a moderate increase of its DNA-binding activity and transactivation, which subsequently protected cells against the tBHP-induced glutathione depletion and cell death. Collectively, our results define a novel modification of Nrf2, which operates as a fine-tuning mechanism for the transcriptional activity of Nrf2 under the oxidative stress.

Effect of phosphorylation and methylation on the function of the p16INK4a protein in non-small cell lung cancer A549 cells.

The p16INK4a protein (p16) has been reported to be a tumor suppressor gene that suppresses the proliferation of cells through the direct inhibition of cell cycle progression. Accordingly, p16 is a potential target for cancer gene therapy. In the present study, the arginine 22, 131 and 138 residues of p16 were found to be methylation sites, as the mutation of these arginine residues to lysine resulted in the hypomethylation of p16. Furthermore, the protein arginine methyltransferases (PRMTs), such as PRMT1, PRMT4 and PRMT6, were determined to be involved in the methylation of the p16 arginine residues. PRMT6 effectively reduced the intensity of the association between p16 and CDK4, and also weakened the function of p16 in preventing cell proliferation. In addition, the p16 protein was found to be phosphorylated in various cell lines, and mutations in the serine residues weakened the cell cycle arrest and induction of apoptosis mediated by p16. Preliminarily, the crosstalk between the phosphorylation and arginine methylation modification of p16 was examined. These findings predict a role for serine phosphorylation against arginine methylation of p16.

A comparison of physicochemical properties of sterilized chitosan hydrogel and its applicability in a canine model of periodontal regeneration.

Chitosan has previously been exploited as a scaffold in tissue engineering processes. To avoid infection, chitosan must be sterilized prior to contact with bodily fluids or blood. Previous research has shown that autoclaved chitosan solution lead to decreased molecular weight, dynamic viscosity, and rate of gelling. We prepared a thermosensitive chitosan hydrogel using autoclaved chitosan powder (121 °C, 10 min) and ß-glycerophosphate (chitosan-PA/GP) and compared the physicochemical properties and biocompatibility in vitro with autoclaved chitosan solution/GP hydrogel. The chitosan-PA/GP hydrogel had a shortened gelation time, higher viscosity, increased water absorption, appropriate degradation time, porous structure, and no obvious cytotoxicity on human periodontal ligament cells. Scanning electron microscopy demonstrated that the cells exhibited a normal morphology. The chitosan-PA/GP hydrogel promoted periodontal tissue regeneration in dog class III furcation defects. The chitosan-PA/GP thermosensitive hydrogel displayed suitable physicochemical properties and biocompatibilities and represents a promising candidate as an injectable tissue engineering scaffold.

[Effect of periodontal initial therapy on teeth with chronic periodontitis and secondary occlusal trauma].

OBJECTIVE: To evaluate the effect of subgingival scaling/root planning (SRP) and occlusal adjustment on clinical and occlusal parameters in teeth with chronic periodontitis and secondary occlusal trauma. METHODS: Eighteen patients with chronic periodontitis and occlusal trauma were included and randomly divided into group A and group B. On day 0, group A was treated by full-mouth subgingival scaling and root planning, and group B was treated by occlusal adjustment in occlusal trauma site. On day 28, group A was treated by occlusal adjustment in occlusal trauma site, and group B was treated by full-mouth subgingival scaling and root planning. Probing depth (PD), attachment loss (AL), bleeding index (BI) were evaluated on 0, 28 and 56 d, and the occlusal time (OT) and the percentage of occlusal force were evaluated on 0, 28 and 56 d in occlusal trauma site. The data was statistically analyzed. RESULTS: In baseline, the PD[(4.42 ± 1.41) mm vs (4.36 ± 1.38) mm], AL [(2.75 ± 1.32) mm vs (2.63 ± 1.37) mm] and BI [(2.20 ± 0.81) vs (2.24 ± 0.89)] of the full-mouth showed no significant difference between the two groups (P > 0.05). There was no significant difference in PD [(5.21 ± 1.21) mm vs (5.08 ± 1.12) mm], AL [(4.94 ± 1.47) mm vs (4.89 ± 1.32) mm], BI [(2.61 ± 0.92) vs 2.50 ± 0.79)], OT [(1.29 ± 0.39) s vs (1.34 ± 0.35) s] and the percentage of occlusal force [(6.8 ± 2.1)% vs (7.4 ± 1.7)%] in occlusal trauma site between the two groups(P > 0.05). After SRP therapy, the PD,AL,BI and OT were significantly decreased (P < 0.05).The clinical parameters exhibited no significant difference after only occlusal adjustment(P > 0.05).On 56 d, the reduction in clinical parameters was not significantly different between the two groups(P > 0.05),however the reduction of OT and the change of the percentage of occlusal force in group A [(0.85 ± 0.41) s, (2.2 ± 2.2)%] were more significant than those in group B [(0.70 ± 0.38) s; (1.5 ± 1.6)%] (P < 0.05). After occlusal adjustment, the increase of OT in group A [(0.21 ± 0.11) s] was lower than that in group B [(0.67 ± 0.37) s]through the 28-day observation period (P < 0.05). CONCLUSIONS: Occlusal adjustment alone is inadequate for control and management of periodontitis.SRP therapy can eliminate the inflammation and decrease the OT of tooth with occlusal trauma.The combination of SRP and occlusal adjustment may achieve more stable results.

Rb protein is essential to the senescence-associated heterochromatic foci formation induced by HMGA2 in primary WI38 cells.

Cellular senescence is an irreversible form of cell cycle arrest that provides a barrier to neoplastic transformation. The integrity of the Rb (Retinoblastoma) pathway is necessary for the formation of the senescence-associated heterochromatin foci (SAHF) that offers a molecular basis for the stability of the senescent state. Surprisingly, although high mobility group A2 protein (HMGA2) can promote tumorigenesis and inhibit Rb function in tumor cells, high-level expression of HMGA2 is sufficient to induce SAHF formation in primary cells. It therefore becomes significant to determine whether Rb protein is necessary in HMGA2-induced SAHF formation. In this study, we established the cellular senescence and SAHF assembly WI38 cell model by ectopic expression of HMGA2, in which typical senescent markers were seen, including notable upregulation of p53, p21 and p16, and elevated SA-ß-galactosidase staining together with downregulation of E2F target genes. We then showed that the Rb pathway inhibitor E7 protein was able to partly abolish the ability of SAHF formation after HMGA2 expression in WI38 cells, indicating that Rb is a crucial factor for HMGA2-induced SAHF formation. However, Rb depletion did not completely rescue the cell growth arrest induced by HMGA2, suggesting that Rb is not an exclusive pathway for HMGA2-induced senescence in WI38 cells.

[Influence of transfection with human transforming growth factor-beta1 gene on the osteogenic potential of the cultured human gingival fibroblast].

OBJECTIVE: To study the influence of transfection with human transforming growth factor-beta1 (hTGF-beta1) gene on the osteogenic potential and differentiation of the cultured human gingival fibroblast (GF). METHODS: Enzyme kinetics method was used to measure the effects of the transfection on the alkaline phosphatase (ALP) activity. Immunohistochemistry stain and image analysis were applied to evaluate the alteration of the content of osteopontin (OPN), bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC) in the GF with transfection. Mineralization nodules formation in vitro was also used. RESULTS: The ALP activity of the GF after transfection was higher than the GF without transfection significantly (P<0.05), and was close to that of the PDLCs (P>0.05). The content of OC in GF was not improved after transfection, was similar with that of PDLCs (P>0.05). Under immunohistochemistry stain, the contents of OPN, ON, BSP expressed in GF with transfection were higher than those of GF without transfection (P<0.05), but similar to those of PDLCs (P>0.05). In the mineralized cultured medium, the nodules were observed in the GF with transfection and PDLCs after 21 days and 24 days alternatively. After von Kossa stain, purple mineralization nodules were observed. CONCLUSION: The GF transfected with pcDNA3-hTGF-beta1 could express some osteogenic characters, though these characters are restricted.

[Effects of gingival fibroblasts transfected with human transform growth factor-beta1 gene on improving the periodontal tissue regeneration].

OBJECTIVE: To evaluate the effects of gingival fibroblasts (GF) transfected with hTGF-beta1 gene on improving the periodontal tissue regeneration for the repair of degree II artificial furcation defects. METHODS: The gingival fibroblasts transfected with hTGF-beta1 gene was compounded to the cuttlebone-transformed nanometer hydroxyapatite (CBHA) material from the cuttlefish in vitro, the degree II furcation defects on the premolars of dogs were produced surgically, and the compound was to implanted into the defect (transfected group), and compared with the compound of periodontal ligament cells (PDLC) with nanometer HA material and the compound of untransfected GF with HA. The results were examined histologically 8 weeks after operation. RESULTS: In the transfected group and the positive control group, more new attachment was found compared with the negative control (P < 0.01), and the NC, NB and NC of the transfected group and the positive control group were: (2.97 +/- 0.50), (4.29 +/- 0.26) and (4.73 +/- 0.06) mm; (3.09 +/- 0.26), (4.46 +/- 0.25) and (4.69 +/- 0.10) mm, respectively. There was no significant difference between the two groups (P > 0.05). Although the alveolar bone regeneration was found in the untransfected group [NB = (3.46 +/- 0.32) mm], the root resorption was observed. The tracing experiment showed that the transfected GF were found in the new alveolar bone and the periodontal membrane. CONCLUSIONS: GF transfected with hTGF-beta1 gene can significantly improve the periodontal tissue regeneration in treatment of degree II furcation defects and is involved in the formation of the new alveolar bone and the new periodontal membrane.