Objective: To investigate the improvement effect of Shenyuan Granule on the vascular calcification in the db/db diabetic nephropathy (DN) mice induced by high phosphorus and its possible mechanism. Methods: Twenty SPF db/db mice were randomly divided into model group(n=10) and Shenyuan Granule group(n=10), and the wild type (WT) mice were chosen as blank group(n=10). After treated for 12 weeks, the serum, kidney tissue and thoracic aorta tissue of the mice in various groups were taken. The serum FGF23 levels of the mice in various groups were detected by ELISA method;HE and von kossa staining were used to detect the pathomorphology of the thoracic aorta and calcium deposition of the mice in various groups; Real-time PCR method was used to detect the expression levels of Klotho mRNA in kidney tissue and Pit-1, Runx2 and SM22α mRNA in thoracic aorta tissue of the mice in various groups; Western blotting method was used to detect the expression levels of Klotho protein in kidney tissue and Pit-1, Runx2 and SM22α proteins in thoracic aorta tissue of the mice in various groups; immunohistochemisty method was used to detect the expression levels of Pit-1 protein in thoracic aorta tissue of the mice in various groups,and immunofluorescence method was used to detect the expression levels of SM22α and Runx2 proteins in thoracic aorta tissue of the mice in various groups. Results: Compared with blank group, the serum FGF23 level of the mice in model group was significantly increased (P<0.05), the expression levels of Klotho mRNA and protein in kidney tissue were significantly decreased (P<0.01), the expression levels of Pit-1 and Runx2 mRNA and proteins in thoracic aorta tissue were increased (P<0.01), the expression levels of SM22α mRNA and protein in thoracic aorta tissue were decreased (P<0.01),the expression levels of Pit-1 and Runx2 proteins in thoracic aorta tissue were increased(P<0.01), and the expression level of SM22α protein in thoracic aorta tissue was decreased (P<0.01).Compared with model group, the serum FGF23 level of the mice in Shenyuan Granule group was decreased (P<0.05), the expression levels of Klotho mRNA and protein in kidney tissue were increased(P<0.01), the expression levels of Pit-1 and Runx2 mRNA and protein in thoracic aorta tissue were decreased (P<0.01), and the expression levels of SM22α mRNA and protein in thoracic aorta tissue were increased (P<0.01). Conclusion: Shenyuan Granule can up-regulate the expression level of Klotho protein in kidney tissue of the mice, reduce the expression level of Pit-1, inhibit the FGF23/Pit-1 signaling pathway, and regulate the phenotypic transformation of vasular smooth muscle cell(VSMC) to achieve its protective effect on the blood vessels.

Objective: To induce the atopic dermatitis (AD) by 2,4-dinitrobenzene (DNCB) in the BALB/c mice,and to explore the possible mechanism of DNCB as hapten to induce AD. Methods: A total of 12 BALB/c mice were randomly divided into control group(n=6) and AD model group (n=6). The dorsal skin of the mice in AD model group was sensitized by 1.0% DNCB at days 1,4,and 7 and the back skin of the left ears were challenged by 0.5% DNCB at days 14,17,19,22,24,27,and 29. The mice in control group were given substrate solution with the same volume at the same time points.The inflammation scores of the skin tissue of the mice in two groups were evaluated, the appearance changes of the ear skin of the mice in two groups were observed, the pathomorphology of the skin tissue in left ear of the mice in two groups were observed by HE staining and toluidine blue staining, the epidermal thickness of the skin tissue at lesion site of the mice in two groups was measured,and the number of mast cells in the skin tissue at lesion site of the mice in two groups was counted; the expression levels of interleukin-4(IL-4) in the skin tissue at lesion site of the mice in two groups was detected by immunohistochemistry staining,and the levels of serum IgE was detected by ELISA method. Results: The inflammation score of the mice in AD model group was higher that in control group(P<0.01).The skin tissue in left ear of the mice in AD model presented as redness swelling and hardening accompaning with abnormal scratching marks;the epidermal thickness, the number of mast cells, the expression level of IL-4 in skin tissue at lesion site and the level of serum IgE of the mice in AD model group were all increased compared with control group(P<0.05 or P<0.01). Conclusion: The AD mouse can be successfully induced by 1.0% DNCB for sensitizing and 0.5% DNCB for challenging in the BALB/c mice,and its mechanism may be related with the increasing of IL-4 expression level in skin tissue and IgE level in serum.

Objective: To construct the RHBDF2 gene over-expression lentivirus vector and to establish the EA.hy926 cells stably expressing RHBDF2, and to provide the evidence for the construction of RHBDF2 gene over-expression lentivirus vector and the establishment of RHBDF2 cells stably expressing RHBDF2. Methods: According to the sequence of RHBDF2 gene provided by NCBI,and the primers were designed and synthesized; the RHBDF2 gene was amplified by PCR method, and the target gene was cloned into the entry vector by Gateway cloning technology, and then subcloned into the lentivirus vector pLV[Exp]-EGFP to construct the recombinant lentivirus plasmid pLV[Exp]-EGFP-RHBDF2; the lentivirus expression vector plasmid pLV[Exp]-EGFP and the recombinant lentivirus plasmid pLV[Exp]-EGFP-RHBDF2 were co-transfected into the HEK293T cells with the virus-assisted packaging plasmids to package the lentivirus and the titer of the lentivirus was detected. The EA.hy926 cells infected with pLV[Exp] - EGFP-control were used as control group and the EA.hy926 cells infected with pLV[Exp] - EGFP-RHBDF2 were used as experiment group. The EA.hy926 cells stably expressing RHBDF2 were screened by puromycin. The fluorescent quantitative PCR (qPCR) and Western blotting methods were used to detect the expression levels of RHBDF2 mRNA and protein in the EA.hy926 cells in control group and experiment group. Results: The enzyme digestion electrophoresis and sequencing results showed that the gene sequence of the EA.hy926 cells over-expression lentivirus vector in experiment group was completely consistent with the designed and synthesized sequence. The lentivirus titer in control group was 1×10⁸ TU·mL^-1, and the lentivirus titer in experiment group was 3×10⁸ TU·mL^-1. The EA.hy926 cells were successfully infected with the lentivirus under fluorescence microscope and the infection efficiency was above 95%.The qPCR detection results showed that the expression level of RHBDF2 mRNA in the EA.hy926 cells in experiment group was higher than that in control group (P<0.01).The Western blotting results showed that the expression level of RHBDF2 protein in the EA.hy926 cells in experiment group was higher than that in control group (P<0.05). Conclusion: The lentivirus vector over-expressing RHBDF2 is successfully constructed, and the EA.hy926 cell line stably up-regulating the expression of RHBDF2 is established by using pLV[Exp]-EGFP-RHBDF2 lentivirus.

Objective: To investigate the effects of macrophage stimulating 1 receptor (MST1R) inhibitor BMS-777607 on the proliferation and apoptosis of the breast cancer MCF-7 cells, and to elucidate the mechanisms. Methods: The breast cancer MCF-7 cells treated with different concentrations of BMS777607 were divided into control group (0μmol·L^-1 BMS-777607 group) and 0.5,1.0,2.0,5.0,10.0,15.0,and 20.0μmol·L^-1 BMS-777607 groups. MTT method was used to detect the proliferation rates of the MCF-7 cells in various groups, and clone formation assay was used to detect the survival rates of the MCF-7 cells in various groups; EDU imaging and EDU flow cytometry methods were used to detect the proliferation rates of the MCF-7 cells in various groups,and Hoechst33342 staining was used to detect the apoptotic morphology of the MCF-7 cells in various groups; flow cytometry was used to detect the apoptotic rates of the MCF-7 cells in various groups,and Western blotting method was used to detect the expression levels of ERK,p-ERK, Akt,p-Akt, PARP,Cleaved PARP, Bax, Caspase-3,Cleaved Caspase-3,Caspase-9 and Cleaved Caspase-9 proteins in the MCF-7 cells in various groups. Results: The MTT results showed that compared with control group, the proliferation rates of the MCF-7 cells in 5 and 10μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01). The clone formation assay results showed that compared with control group, the survival rates of the MCF-7 cells in 5 and 20μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01).The EDU incorporation results showed that compared with control group, the proliferation rates of the MCF-7 cells in 10 and 20μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01).The Hoechst33342 fluorescence staining results showed that the MCF-7 cells in control group showed the light staining, a small number of MCF-7 cells in 10μmol·L^-1 BMS-777607 group showed the bright nuclei,and most of the MCF-7 cells in 20μmol·L^-1 BMS-777607 group showed the bright nuclei. The flow cytometry results showed that compared with control group, the apoptotic rates of the MCF-7 cells in 10 and 20μmol·L^-1 BMS-777607 groups were decreased (P<0.05). The Western blotting results showed that compared with control group, the expression levels of p-ERK and p-Akt proteins in the MCF-7 cells in 10, 15, and 20μmol·L^-1BMS-777607 groups were significantly decreased(P<0.05 or P<0.01), and the expression levels of PARP, Cleaved PARP, Bax, Cleaved Caspase-3, and Cleaved Caspase-9 proteins were significantly increased(P<0.05 or P<0.01). Conclusion: MST1R inhibitor BMS777607 can inhibit the proliferation of MCF-7 cells and induce the apoptosis, and their mechanisms are related to the inhibition of p-ERK and p-Akt expressions and the promotion of expressions of Cleaved PARP, Bax, Cleaved Caspase-9 and Cleaved Caspase-3.

Objective: To use Escherichia coli (E. coli)to prepare the recombinant proteins of elastin-like polypeptide(ELP) for displaying the two epitopes of N-terminal pro-brain natriuretic peptide(NT-proBNP), and to provide the basis for the low-cost and high-efficiency preparation of NT-proBNP detection calibrator. Methods: The epitopes of 13-20 and 63-71 amino acid residues of NT-proBNP were designed and fused with ELP by flexible chain;three kinds of fusion proteins were obtained. The genes encoding the three fusion proteins mentioned above were synthesized by genetic engineering technique and cloned into the pET-28a (+) vector; the recombinant proteins were induced and expressed automatically in E.coli BL21(DE3),the recombinant proteins were purified with inverse transition cycling(ITC),and the abilities of their antibodies specificly binding epitopes were detected by Western blotting and ELISA methods. Results: Three kinds of vectors of ELP for displaying NT-proBNP epitopes were successfully constructed, and the corresponding recombinant proteins were expressed in the E.coli. The western blotting and direct ELISA results showed that the specific epitopes displayed by ELP had the better binding ability with the relative antibodies.The results of Sandwich ELISA showed that the protein concentrations of recombinant proteins of ELP for displaying two epitopes of NT-proBNP had a double logarithmic linear dose-dependent relationship with the absorbance (A) value at 450 nm (r=0.9197, P<0.01). Conclusion: The ELP is successfully used to display the two epitopes of NT-proBNP, and lay a foundation for the low-cost and high-efficiency preparation of the NT-proBNP detection calibrator.

Objective: To investigate the changes of expressions of the related genes in the triple-negative breast cancer(TNBC) MDA-MB-231 cells after knockout of Rab7a gene, and to elucidate the roles of Rab7a-related genes in TNBC. Methods: The breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231 and HCC-1937 cells in the logarithmic phase were selected.The expression levels of Rab7a protein in the breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231, and HCC-1937 cells were detected by Western blotting method. The Rab7a gene in the MDA-MB-231 cells was knockout with lentivirus and the cells were divided into negative control group and Rab7a knockout group. According to the four different knockout Rab7a sequences, the Rab7a knockout group was divided into KD1 group, KD2 group, KD3 group and KD4 group. The knockout efficiencies of Rab7a gene in the MDA-MB-231 cells in various groups were detected by qPCR method, the differential genes in the highest knockout efficiency group (KD2 group) and negative control group were detected by full gene expression microarray,and the interaction between Rab7a gene and other genes was analyzed by integrated pathway analysis (IPA) software. Results: The Rab7a protein was expressed in the TNBC MDA-MB-231 cells. Compared with negative control group, the knockout efficiency of Rab7a gene in the MDA-MB-231 cells in KD2 group was the highest (P<0.01); compared with negative control group, there were 634 differential genes in the MDA-MB-231 cells in KD2 group, the number of up-regulated genes and down-regulated genes were increased(P<0.01); the differential gene analysis results showed that Rab7a knockout had the relationships with cancer, cell survival, cell cycle, cell growth,and migration. The expressions of eIF4F, IRS1 and RPS6KB1 in the Rab7a IPA network diagram were down-regulated, and the expressions of PRKAA1, IKBKE, VEGFA,and transcription factor ATF2 were up-regulated. Conclusion: Rab7a gene plays a role as a gene network in the TNBC MDA-MB-231 cells. The Rab7a gene interacts with the multiple genes and participates in the pathological processes of cancer cells.

Objective: To investigate the changes of expressions of tubulin and endosome-lysosome in the neurons in hippocampus tissue of the mice after status epilepticus (SE), and to elucidate the change rule of microtubule and endosome-lysosome system in the process of delayed neuronal death. Methods: A total of 40 male ICR mice were divided into control group(n=7,given normal saline) and experiment group(n=33, give pilocarpine);the mice in experiment group met the SE standand were divided into SE 1 d, SE 2 d, SE 3 d and SE 7 d groups according to the time after SE(n=5). Nissl and Fluoro-Jade B (F-JB) staining methods were used to detect the damage of neurons in hippocampus tissue of the mice in various groups. The expression intensities of β-tubulin,endosom protein Rab5 and lysosome constitutive protein LAMP1 and the percentages of β-tubulin, Rab5 and LAMP1 positive areas in neurons in hippocampus tissue of the mice in various groups were detected by immunofluorescence method. The relationships between the expression of β-tubulin and the expressions of Rab5 and LAMP1 in hippocampal CA1 area of the mice in various groups were detected by double fluorescence method. Results: Compared with control group, the number of neurons in hippocampal CA1 and CA3 areas of the mice in SE 1 d group was decreased (P<0.01), the number of F-JB positive cells was increased (P<0.01); the number of Nissl positive neurons in hippocampus tissue of the mice in SE 2 d, SE 3 d, and SE 7 d groups was decreased (P<0.01). Compared with control group, the number of F-JB positive neurons in hippocampus tissue of the mice in SE 2 d,SE 3 d and SE 7 d groups was increased (P<0.01). Compared with control group, the percentages of β-tubulin positive areas in hippocampal CA1 and CA3 areas of the mice in SE 2 d, SE 3 d and SE 7 d groups were significantly decreased (P<0.05), and the trend of change was similar to that of neuron damage. Compared with control group, the expression intensities of Rab5 and LAMP1 in neurons in hippocampus tissue of the mice in SE 1 d, SE 2 d, SE 3 d and SE 7 d groups were decreased with the prolongation of time, the percentages of Rab5 positive areas were significantly decreased (P<0.05 or P<0.01),and the percentages of LAMP1 positive areas were increased in a short time on the first day after SE and decreased with the prolongation of time(P<0.05 or P<0.01). The double fluorescence staining results showed that 1 d after SE was the key point for the decrease of endosome and the transient increase of lysosome in the early stage, which was consistent with the occurrence time of microtubule injury. Conclusion: SE can cause delayed neuronal death, at the same time, the tubulin skeleton of neurons is damaged, and the location and function of endosome-lysosome system are also changed abnormally.

Objective: To investigate the inhibitory effect of mitochondrial dynamics changes on the tumor growth of transplanted melanoma in the APP/PS1 the mice and its mechanism. Methods: The male C57BL/6J(C57) and APP/PS1 mice were divided into C57 transplantation tumor group (n=7) and APP/PS1 transplantation tumor group(n=7). The tumor appearance time was observed and the tumor volume of the mice in two groups were calculated and the growth curves of the mice in two groups were drawn. The morphology of tumor tissue of the mice in two groups was observed under light microscope,the expression levels of mitofusion 2(Mfn2), dynamin related protein (Drp1), fission 1(Fis1), ubiquitin and polyubiquitin, LC3-Ⅱ, PTEN induced putative kinase 1(PINK1) and Parkin proteins in tumor tissue of the mice in two groups were detected by Western blotting method. Results: The tumor tissue under the skin of the mice in C57 transplantation tumor group and APP/PS1 transplantation tumor group was found with the melanoma granules in the tumor cells. Compared with C57 transplantation tumor group, the tumor appearance time of the mice in APP/PS1 transplantation tumor group was late, and the tumor volume was decreased (P<0.05); the expression level of Mfn2 protein in tumor tissue of the mice was decreased(P<0.05); the expression levels of Drp1 and Fis1 proteins were increased (P<0.05), and the expression levels of ubiquitin and polyubiqitin, LC3-Ⅱ, PINK1 and Parkin proteins in tumor tissue of the mice in APP/PS1 transplantation tumor group were all increased(P<0.05). Conclusion: The growth of tumor in the APP/PS1 mice with transplanted melanoma is slow,and its mechanism may be related to PINK1/Parkin pathway involved in the mitochondrial autophagy.

Objective: To investigate the effect of microRNA-9-5p (miR-9-5p) targeting the myocyte enhancer factor 2C (MEF2C) on the biologicals behaviors of the alveolar rhabdomyosarcoma (ARMS) cells, and to provide the basis for the molecular diagnosis and targeted therapy of ARMS. Methods: The expression levels of miR-9-5p and MEF2C mRNA in ARMS tissue and cells were detected by qRT-PCR method, the proliferation rate of cells was detected by CCK-8 method, the apoptotic rate was detected by flow cytometry, the numbers of invasion and migration cells were detected by Transwell chamber assay, the luciferase activity in 293T cells was detected by double luciferase reporter gene, and the expression level of MEF2C protein in the cells was detected by Western blotting method. Results: The expression levels of miR-9-5p in ARMS tissue and cells were higher than those in normal skeletal muscle tissue and HSKMC cells (P<0.05). Compared with miR-NC group, the expression level of miR-9-5p in RH30 cells in miR-9-5p inhibitor group was decreased (P<0.01),the proliferation rate of RH30 cells in miR-9-5p inhibitor group was decreased (P<0.05), the apoptotic rate was increased (P<0.05), and the numbers of invasion and migration cells were decreased (P<0.05). The luciferase activity in the 293T cels after co-transfection with MEF2C-3'-UTR-WT and added with miR-9-5p was decreased (P<0.01);compared with miR-NC group, the expression levels of MEF2C mRNA and protein in the RH30 cells in miR-9-5p inhibitor group were increased (P<0.05).The expression level of MEF2C mRNA in the ARMS tissue was lower than that in normal skeletal muscle tissue (P<0.01), which was negatively correlated with the expression of miR-9-5p in ARMS tissue (r=-0.5420, P<0.05);the expression level of MEF2C mRNA in the RH30 cells and PLA802 cells was lower than that in the HSKMC cells (P<0.01).Compared with transfection with control plasmid(EV), the expression level of MEF2C mRNA in the RH30 cells after transfection with MEF2C over-expression plasmid was increased (P<0.01), the proliferation rate was decreased (P<0.01), the apoptotic rate was increased (P<0.05), the number of invasion cells was decreased (P<0.05), and the number of migration cells was decreased (P<0.01).Compared with transfection with si-NC, the expression level of MEF2C mRNA in the RH30 cells after transfection with MEF2C siRNA was decreased (P<0.01), the proliferation rate was increased (P<0.05), the apoptotic rate was decreased (P<0.01), the number of invasion cells was increased (P<0.01),and the number of migration cells was increased (P<0.01). Conclusion: MiR-9-5p can directly target the 3'-UTR of MEF2C mRNA to induce mRNA degradation and inhibit the expression of MEF2C, and promote the proliferation, invasion, migration and anti-apoptosis ability of the RH30 cells.

Objective: To observe the effect of type Ⅰbone morphogenetic protein(BMP)receptor activin A receptor type Ⅰ(ACVR1)on the morphology,proliferation and differentiation of the mandibular condylar cartilage (MCC) cells in the postnatal mice, and to provide the reference for the study on etiology and treatment of MCC-related disease. Methods: The C57BL/6J mouse model of conditional deletion of ACVR1 gene was constructed by using the Cre-LoxP system. The female and male mice with Acvr1^fx/fx; RS/RS and Acvr1^+/-; Osterix (+)/(-) genotypes were paired off with each other; the offspring Osterix-Cre (+); Acvr1^fx/-; RS/+ genotype mice were selected as experiment group,and the Osterix-Cre (+); Acvr1^fx/+; RS/+ mice were selected as control group. The newborn (n=3), postnatal day 21(PN21) (n=4) and PN42(n=5) male mice were selected. X-gal staining was used to detect the expressions of Osterix-Cre in MCC tissue of the mice in two groups,micro-CT was used to detect the condylar widths and condylar head lengths of mandible of the mice in two groups,HE and Toluidine blue staining were used to analyze the morphology of MCC cells and the thickness of caritilage in each layer of MCC tissue of the mice in two groups,immunohistochemical (IHC) staining was used to detect the number of proliferating cell nuclear antigen(PCNA)-positive cells and the level of type Ⅹ collagen in MCC tissue of the mice in two groups. Results: The X-gal staining and IHC results showed that the mouse model of ACVR1 gene conditional deletion was successfully constructed. At PN21, compared with control group, the condylar width and the condylar head length of mandible of the mice in experiment group were significantly shortened (P<0.05); the morphology of the MCC cells of the mice in two groups had no significant difference. Compared with control group, the number of PCNA-positive cells in the MCC cells of hypertrophic chondrocyte zone(Hy) and chondroblastic zone(Ch) and single Hy of the mice in experiment group were significantly increased(P<0.05 or P<0.01).At PN42, compared with control group, the shape of parts of the mandibular condylar cartilage cells of the mice in experiment group was abnormal, and the arrangement of some condylar chondrocytes was disordered, the cell thickness of the Ar, Pr and Ch in intermediate part and Hy in anterior part of the condylar cartilage of the mice in experiment group were significantly increased(P<0.05 or P<0.01); compared with control group, the number of PCNA-positive cells in each zone and the level of type Ⅹ collagen in Ch of MCC tissue of the mice in experiment group were incresed. Conclusion: ACVR1 affects the morphology of MCC cells and structure of MCC tissue by inhibiting the proliferation of MCC cells and the differentiation of chondroblasts into hypertrophic chondrocytes.

Objective: To investigate the effect of baicalin on the autophagy of the synovial RSC-364 cells of the rats induced by lipopolysaccharide (LPS)through PI3k/Akt/mTOR signal pathway, and to clarify its mechanism. Methods: The rat synovial RSC-364 cells in logarithmic growth phase were divided into control group, model group, dexamethasone(DXMS) group, 10μmol·L^-1baicalin group, 20μmol·L^-1 baicalin group and 40μmol·L^-1 baicalin group. The RSC-364 cells in control group were only supplemented with culture medium, and the RSC-364 cells in the other groups were stimulated with 1 mg·L^-1LPS for 12 h to make the inflammatory cell models. The survival rates of RSC-364 cells were detected by MTT assay, and the expression levels of PI3k, Akt, mTOR, Beclin1, and LC3-Ⅱ mRNA in the RSC-364 cells in various groups were detected by RT-PCR method;Western blotting method was used to detect the expression levels of Beclin1, Atg5, Atg7, Atg12, microtubule-associated protein-light chain 3-Ⅱ(LC3-Ⅱ), and P62 proteins in the RSC-364 cells in various groups. Results: Compared with control group, the survival rate of RSC-364 cells in model group was significantly increased (P<0.01), the expression levels of PI3k, Akt, mTOR mRNA and P62 protein in the RSC-364 cells in model group were significantly decreased (P<0.05 or P<0.01), and the expression levels of Atg5, Atg7, Atg12, LC3-Ⅱ, Beclin1 mRNA and proteins were significantly increased (P<0.01); compared with model group, the survival rates of RSC-364 cells in different concentrations of baicalin groups were significantly decreased (P<0.05 or P<0.01), the expression levels of PI3k, Akt, mTOR mRNA and P62 protein in the RSC-364 cells in different concentrations of baicalin groups were significantly increased (P<0.05 or P<0.01), and the expression levels of Atg5, Atg7, Atg12, LC3-Ⅱ and Beclin1 mRNA and proteins were decreased (P<0.05 or P<0.01). Conclusion: Baicalin may inhibit the LPS-induced autophagy of the RSC-364 cells by activating the PI3k/Akt/mTOR signaling pathway.

Objective: To explore the establishment of the incisor extraction wound models in aesthetic area of the rats and the application effect of modified minimally invasive tooth extraction surgery, and to provide the basis for establishment of the incisor extraction wound models in aesthetic area and improvement of the healing of alveolar bone and soft tissue. Methods: Twenty-four SD rats were randomly divided into control group and experiment group (n=12). The crowns of right lower incisors of the rats were removed along with the top of gingival tissue on the 9th, 6th and 3rd days before the incisors extraction, and then the incisor were extracted on the 10th day of experimental modeling. In experiment group, the incisors of rats were extracted by the modified minimally invasive incisor extraction, while in control group the conventional extraction was used to extract the incisor of the rats. The survival rates and body weights of the rats in two groups were observed before and after the incisor extraction. The complete degrees of incisor extraction of the rats in two groups were detected,and the rates of complete incisor extraction were calculated; the healing condition of soft tissue around the incisor extraction of the rats in two groups 1 week after incisor extraction and the recovery condition of mandibular alveolar bone of the rats 4 weeks after incisor extraction were compared. Results: The survival rate of the rats in experiment group was higher than that in control group (P<0.05). The body weights of the rats in experiment group at different time points after extraction were higher than those in control group (P<0.05). Compared with control group, the rate of complete incisor extration of the rats in experiment group was increased(P<0.01).The soft tissue around the incisor extraction wound of the rats in experiment group was completely healed within one week after incisor extraction, and new bone was found at the top of alveolar ridge four weeks after incisor extraction; meanwhile, the soft tissue around the incisor extraction wound of the rats in control group was still red and swollen one week after incisor extraction, and the trabecular structure of bone at the top of alveolar ridge was disordered found weeks after incisor extracion, and the new bone structure was poor. Conclusion: The modified minimally invasive tooth extraction surgery can significantly improve the complete extraction effect of incisors in the rats, and the soft tissue around the incisor extraction wound and the alveolar bone are healed well after minimally invasive tooth extraction.

Objective: To investigate the effects of polyactic-co-glycolic(PLGA) electrospinning membrane carrying paclitaxel-liposomes with different concentrations on the proliferation and differentiation of the neural stem cells (NSCs), and to evaluate the feasibility of PLGA electrospinning membrane carrying paclitaxel liposomes in the tissue engineering repair of spinal cord injury. Methods: The paclitaxel-liposomes and PLGA were mixed in different proportions and the electrospinning technique was used to construct 1, 5 and 10μg·L^-1electrospinning membrane carrying paclitaxel-liposomes. The NSCs were isolated and purified from the fetal rat brain tissue. The diameters of PLGA electrospinning membrane carrying paclitaxel-liposomes in various groups were detected by scanning electron microscope (SEM), and the isolated and purified NSCs were identified by immunofluorescence staining. The NSCs were cultured in PLGA electrospinning membrane carrying paclitaxel-liposomes with different concentrations (0,1,5 and 10μ g·L^-1)as PLGA electrospinning membrane group and PLGA electrospinning membrane carrying 1,5 and 10μ g·L^-1paclitaxel-liposomes groups. The proliferation levels of the NSCs in various groups was detected by MTT method, and the expression levels of Tuj-1 and GFAP mRNA in the NSCs in various groups were detected by RT-PCR method. Results: The PLGA electrospinning membrane carrying paclitaxel-liposomes were white thin films. The SEM results showed that there was no statistical differences in the fiber diameters of PLGA electrospinning membrane carrying paclitaxel-liposomes between various groups (P>0.05).The immunofluorescence staining results showed that the cells isolated from fetal rat brain tissue were spherical and positive expression of Nestin protein. The MTT assay results showed that the proliferation level of NSCs in PLGA electrospinning membrane carrying 5μg·L^-1paclitaxel-liposomes group was higher than those in the other three groups(P<0.05).The RT-PCR results showed that compared with PLGA electrospinning membrane group, the expression level of Tuj-1 mRNA in the NSCs in PLGA electrospinning membrane carrying 5μg·L^-1 paclitaxel-liposomes group was increased(P<0.05) and the expression level of GFAP mRNA in the NSCs in PLGA electrospinning membrane carrying 5μg·L^-1 paclitaxel-liposomes group was decreased(P<0.05). Conclusion: Medium concentration of paclitaxel can promote the proliferation of NSCs and induce the differentiation of NSCs into the neurons. PLGA electrospinning membrane carrying medium concentration of paclitaxel-liposomes has certain application value in the repair of spinal cord injury.

Objective: To synthesize the zinc-doped carbon dots (CDs) by hydrothermal method and to observe the inhibitory effects of zinc-doped CDs combined with blue light on the formation of Staphylococcus aureus(S.aureus),and to explore the related mechanism. Methods: The zinc-doped(CDs) were synthesized by hydrothermal method, and the characteristics were observed by transmission electron microscope (TEM), fluorescence spectrometer and Fourier transform-infrared spectrometer (FT-IR). The experiment was divided into blank control group, CDs group, blue light radiation group,and CDs +blue light radiation group. The cells in CDs group were treated with different concentrations (50, 75, 100 mg·L^-1) of CDs, the cells in blue light radiation group were irradiated with blue light for 10, 20,and 40 min, the cells in CDs + blue light radiation group were treated with CDs combined with blue light,and the cells in blank control group were only treated by culture medium in the dark.CCK-8 assay was used to determine the proliferation rates of the L929 and MC3T3-E1 cells. The reactive oxygen species were scavenged by adding N-acetylcysteine(NAC) in the best bacteriostatic effect group(100 mg·L^-1 CDs group),and the experiment was divided into contro group,100 mg·L^-1 CDs group,0.5 mmol·L^-1NAC group,and 0.5 mmol·L^-1 NAC+100 mg·L^-1 CDs group. The concentrations of the bacteria suspension in various groups were detected by spectrophotometer,the bacterial biofilm formation amounts of S.aureus in various groups were detected by crystal violet staining,and the plate count method was used to record the colony counts in various groups. Results: The TEM results showed that the particle size of the zinc-doped CDs constructed successfully was about 1.8 nm. The fluorescence spectra showed that the optimum excitation wavelength of CDs was 342 nm and the optimum emission wavelentgh was 450 nm. The FT-IR spectrum showed that CDs had hydroxyl, carboxy, amino and other functional groups. The CCK-8 assay results showed that after co-culture for 24 h,the proliferation rates of L929 and MC3T3-E1 cells in 100 mg·L^-1 CDs group were up to 80%. Compared with blank control group, the concentrations of bacteria solution in blue light radiation for 20 and 40 min groups were decreased (P<0.05 or P<0.01), the biofilm formation amount of S.aureus was decreased after blue light radiation for 40 min(P<0.01).Compared with blue light radiation group, the concentration of bacteria solution and the biofilm formation amount of S.aureus in CDs + blue light radiation(10 min) group were decreased after blue light radition for 10 min (P<0.01).Compared with 100 mg·L^-1 CDs group, the concentration of bacteria solution and the biofilm formation amount of bacteria in 0.5 mmol·L^-1 NAC+100 mg·L^-1 CDs group were increased (P<0.01). Conclusion: Zinc-doped CDs combined with blue light can inhibit the growth of S.aureus and the biofilm formation by photocatalysis effectly.

Objective: To observe the inhibitory effect of combination of disulfiram (DSF) and cisplatin (CDDP) on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells, and to explore its possible mechanism. Methods: The MDA-MB-231 cells in the logarithmic phase were divided into blank control group, DSF group, CDDP group and combination groups (60μmol·L^-1 CDDP+1,2 and 4μmol·L^-1DSF). MTT method was used to detect the survival rates of the MDA-MB-231 cells in various groups and flow cytometry was used to detect the apoptotic rates of the MDA-MB-231 cells in various groups, flow cytometry was used to detect the level of active oxygen(ROS)in the MDA-MB-231 cells in various groups, and Inductively Coupled Plasma-MassSpectrometry (ICP-MS) was used to detect the levels of Pt in the MDA-MB-231 cells in various groups. Results: The MTT method and flow cytometry results showed that after treated for 72 h, compared with 60μmol·L^-1 CDDP group,the survival rates of the MDA-MB-231 cells in combination 1 group (60μmol·L^-1CDDP+1μmol·L^-1DSF), combination 2 group (60μmol·L^-1CDDP+2μmol·L^-1DSF) and combination 3 group (60μmol·L^-1CDDP+4μmol·L^-1DSF) were decreased(P<0.05). After treated for 24 h, compared with 60μ mol·L^-1CDDP group and 2μ mol·L^-1DSF group, the apoptotic rate of the MDA-MB-231 cells in combination group (60μmol·L^-1CDDP+2μmol·L^-1 DSF) was increased (P<0.05). Compared with CDDP group, the level of ROS in the MDA-MB-231 cells in combination group(60μmol·L^-1CDDP+2μmol·L^-1 DSF) was increased (P<0.05).The ICP-MS results showed that compared with CDDP group, the level of Pt in the MDA-MB-231 cells in combination group(20μmol·L^-1CDDP+0.625μmol·L^-1 DSF) was increased (P<0.05). Conclusion: The combination of DSF and CDDP can significantly inhibit the proliferation of MDA-MB-231 cells,and its possible mechanism may be related to inhibiting the growth of tumor cells by increasing the ROS level and Pt content in the MDA-MB-231 cells.

Objective: To investigate the killing effect of amplified natural killer (NK) cells on the gastric cancer cells,and to elucidate its mechanism. Methods: The peripheral blood mononuclear cells (PBMCs) from 15 patients with gastric cancer were extracted and isolated. The morphology of NK cells before and after amplification was observed, the percentages of NK cells before and after amplification were detected, and the amplification time of NK cells after amplification was calculated.The killing effects of NK cells on the gastric cancer cells before and after amplification were detected. The percentages of expressions of killing activating receptors NKG2D and DNAM-1 and killing inhibitory receptors KIR2DL1 and KIR3DL1 were detected by flow cytometry. Results: Before amplification, the NK cells were round, small in size and scattered in distribution. After amplification, the NK cells were increased in size and irregular in shape. The percentage of NK cells after amplification was significantly higher than that before amplification (P<0.01), and the number of the NK cells after amplification was (596±152) times of before amplification. When the effective target ratio was 5:1, the killing activity of NK cells on the gastric cancer cells after amplification was significantly higher than that before amplification (P<0.01).After amplification, the percentages of expressions of killing activating receptors NKG2D and DNAM-1 were significantly higher than those before amplification (P<0.01).After amplification, the percentages of expressions of killing inhibitory receptors KIR2DL1 and KIR3DL1 were significantly lower than those before amplification (P<0.05). Conclusion: The killing effect of NK cells on the gastric cancer cells after amplification is stronger than before amplification. The mechanism may be related to increasing the expressions of activated receptors and decreasing the expressions of inhibitory receptors on the surface of NK cells after amplification.

Objective: To investigate the effect of interleukin-4(IL-4) and estradiol on the biological behavior of breast cancer 4T1 cells of the mice, and to elucidate its mechanism. Methods: The 4T1 cells were cultured in vitro and added with different concentrations (0, 12.5, 25.0, 50.0 and 100.0μg·L^-1) of IL-4 or estradiol (0, 6.25, 12.50, 25.00 and 50.00 nmol·L^-1).The proliferation rate of the breast cancer 4T1 cells was measured by MTT method after treated for 72 h.The breast cancer 4T1 cells were divided into control group (without any treatment), IL-4 group (treated with 50.0μg·L^-1IL-4), estradiol group (treated with 12.50 nmol·L^-1 estradiol) and combination group (treated with 50.0μg·L ^-1 IL-4+ 12.50 nmol·L^-1 estradiol). MTT method was used to detect the proliferation rates of the breast cancer 4T1 cells in various groups, and flow cytometry was used to detect the percentages of the breast cancer 4T1 cells in different cell cycles in various groups,and Western blotting method was used to detect the expression levels of STAT6, p-STAT6, ERα, Erk, p-Erk, P70S6K, p-P70S6K, S6,and p-S6 in the breast cancer 4T1 cells in various groups. Results: Compared with 0μg·L^-1 IL-4 group, the proliferation rates of the breast cancer 4T1 cells in 25.0, 50.0 and 100.0μg·L^-1 IL-4 groups were increased (P<0.05);compared with 0 nmol·L^-1 estradiol groups, the proliferation rates of the breast cancer 4T1 cells in 12.50, 25.00 and 50.00 nmol·L^-1 estradiol groups were increased (P<0.05).Compared with control group, the proliferation rate of the breast cancer 4T1 cells in IL-4 group was increased (P<0.05); compared with control group, the proliferation rate of the breast cancer 4T1 cells in estradiol group was increased (P<0.05); compared with IL-4 group or estradiol group, the proliferation rate of the breast cancer 4T1 cells in combination group was increased (P<0.05).Compared with control group, the percentages of the breast cancer 4T1 cells at S phase and G₂/M phase in IL-4 group were increased (P<0.05),and the percentage of the breast cancer 4T1 cells at G₀ and G₁ phases were decreased(P<0.05); compared with control group, the percentage of the breast cancer 4T1 cells at S phase in estradiol group was increased(P<0.05),and the percentages of the breast cancer 4T1 cells at G₀ and G₁ phases were decreased (P<0.05).Compared with control group, the expression levels of ERα, p-Erk, p-P70S6K, and p-S6 in the breast cancer 4T1 cells in IL-4 group were increased(P<0.05), while the expression levels of p-STAT6, ERα, p-Erk, p-P70S6K, S6,and p-S6 in the breast cancer 4T1 cells in estradiol group were increased (P<0.05); the expression levels of STAT6, p-STAT6, ERα, p-ERK, p-P70S6K, and p-S6, in the breast cancer 4T1 cells in combination group were increased (P<0.05). Conclusion: The combination of IL-4 and estradiol can increase the expressions of IL-4 receptor(IL-4R) and estrogen receptor (ER),and enhance the activation of Erk1, p70S6K kinase and phosphorylation of downstream S6 protein in the breast cancer 4T1 cells.

Objective: To investigate the effects of lysine acetyltransferase (Kat5) on the proliferation and apoptosis of the thyroid papillary carcinoma cells and the role of phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinase (AKT) signaling pathway,and to elucidate the possible mechanism of Kat5 in the proliferation and apoptosis of thyroid papillarycarcinoma cells. Methods: RT-PCR and Western blotting methods were used to determine the expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3, TPC-1, K1 cells and thyroid epithelial Nthy-ori3-1 cells.The thyroid papillary carcinoma TPC-1 cells were divided into blank control group, negative control group and silencing Kat5 group (si-Kat5 group).The cells in blank control group were not transfected,the cells in negative control group were transfected with negative control siRNA,and the cells in si-Kat5 group were transfected with Kat5 siRNA.The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma TPC-1 cells in various groups were detected by RT-PCR and Western blotting methods.The proliferation activities of the TPC-1 cells in various groups were measured by CCK-8 method,the colony formation assay was used to measure the clone formation rates of the TPC-1 cells in various groups,the apoptotic rates of the TPC-1 cells in various groups were detected by flow cytometry,and the expression levels of cyclin-dependent kinase 2(CDK2), P21, P53, cleaved caspase-3, PI3K, phosphorylated PI3K (p-PI3K), AKT, and phosphorylated AKT(p-AKT) protein in the TPC-1 cells in various groups were detected by Western blotting method. Results: The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3, TPC-1 and K1 cells were higher than those in thyroid epithelial Nthy-ori3-1 cells (P<0.05).The differences in the Kat5 mRNA and protein expression levels,the proliferation activities,the clone formation rates,the apoptotic rates and the expression levels of CDK2, P21, P53, cleaved caspase-3, PI3K, p-PI3K, AKT and p-AKT proteins in the TPC-1 cells between various groups were statistically significant(P<0.05).Compared with blank control group and negative control group, the expression levels of Kat5 mRNA and protein in the TPC-1 cells in si-Kat5 group were decreased(P<0.05), the proliferation activity was decreased(P<0.05),the colony formation rate was decreased(P<0.05),the apoptotic rate was increased(P<0.05), the expression levels of CDK2, p-PI3K and p-AKT proteins were decreased(P<0.05), and the expression levels of P21, P53 and cleaved caspase-3 proteins were increased(P<0.05). Conclusion: The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma cells are increased. Silencing Kat5 gene can inhibit the proliferation of the thyroid papillary carcinoma cells and promote their apoptosis,and tis mechanism may be related to the inhibition of PI3K/AKT signaling pathway.

Objective: To investigate the expressions of Galectin-3 and matrix metalloproteinase-9(MMP-9) in brain tissue of the mice with acute cerebral infarction and the intervention effect of nimodipine,and to elucidate the effects of Galectin-3 and MMP-9 in the occurrence and treatment of cerebral infarction. Methods: A total of 210 mice were randomly divided into control group, model group and nimodipine group (n=70).In model group and nimodipine group, the acute cerebral infarction models of mice were established by the suture method.The mice in nimodipine group were intraperitoneally injected with nimodipine 30 min before operation.The Longar score and modified neurological severity score (mNSS) were used to evaluate the neurological function of the mice in various groups,the cerebral infarction volumes of the mice in various groups were determined by TTC staining,the water contents in brain tissue of the mice in various groups were detected, HE staining was used to observe the morphology of brain tissue of the mice in various groups, the expression levels of Galectin-3 and MMP-9 mRNA in brain tissue of the mice in various groups were detected by RT-PCR method,and the expression levels of Galectin-3 and MMP-9 proteins in brain tissue of the mice in various groups were detected by immunohistochemical staining. Results: Compared with control group, the Longar score and mNSS of the mice in model group were increased(P<0.05),the cerebral infarction volume and water content in brain tissue of the mice in model group were significantly increased(P<0.05), the expression levels of Galectin-3 and MMP-9 mRNA in brain tissue were increased(P<0.05),and the expression levels of Galectin-3 and MMP-9 proteins in brain tissue were increased(P<0.05).Compared with model group, the Longar score and mNSS of the mice in nimodipine group were decreased (P<0.05),the cerebral infarction volume and the water content in brain tissue were decreased(P<0.05),the expression levels of Galectin-3 and MMP-9 mRNA in brain tissue were decreased(P<0.05), and the expression levels of Galectin-3 and MMP-9 proteins were decreased(P<0.05).The morphology and structure of brain cells of the mice in control group were normal;the brain cells of the mice in model group showed edema and flaky necrosis, and the inflammatory cell infiltration was found; the brain cells of the mice in nimodipine group showed mild edema and necrosis. Conclusion: Nimodipine can protect the brain tissue of the mice with cerebral infarction by reducing the expressions of Galectin-3 and MMP-9.

Objective: To investigate the protective effect of Shuxuening Injection on the brain tissue of the rats with acute cerebral infarction,and to elucidate its mechanism. Methods: Fifty rats were randomly divided into control group, sham operation group, model group, nimodipine group and Shuxuening Injection group(n=10).The rat models of acute cerebral infarction were made by internal carotid artery suture method in model group, nimodipine group and Shuxuening Injection group. The common carotid arteries, the external carotid arteries and the internal carotid arteries of the rats in sham operation group were separated,and only the external carotid arteries were ligated; the rats in control group were not treated with operation; the rats in Shuxuening Injection group were given Shuxuening Injection, the rats in nimodipine group were given nimodipine,and the rats in control group, model group and sham operation group were given normal saline at the same volume.The score of neurological impairment, water contents in brain tissue and relative cerebral infarction areas of the rats in various groups were measured.HE staining was used to observe the morphology of brain tissue of the rats in various groups,and immumohistochemical staining was used to detect the expression levels of growth differentiation factor -15(GDF-15) and C-reactive protein (CRP) in brain tissue of the rats in various groups. ELISA method was used to detect the levels of tumor necrosis factor-α(TNF-α), interleukin- 6(IL-6)and interleukin-1β(IL-1β)in brain tissue of the rats in various groups. Results: The cortical structures of cortex of the rats in control group and sham operation group were complete and the cells were arranged neatly;the cytoplasm and nucleus of the nerve cells of the rats in model group were wrinkled, and the interstitium between nerve cells and capillaries were loose; the swelling degrees of cerebral cortical nerve cells and glial cells and the loose degrees of stroma of the rats in Shuxuening Injection group were reduced, and the histopathological manifestations were similar to those in nimodipine group. Compared with control group and sham operation group, the water content in brain tissue of the rats in model group was significantly increased(P<0.05), and the expression levels of CRP,the levels of TNF-α, IL-6 and IL-1β in brain tissue of the rats in model group were significantly increased(P<0.05), while the expression level of GDF-15 was significantly decreased(P<0.05).Compared with model group, the scores of neurological impairment of the rats in nimodipine group and Shuxuening Injection group were significantly decreased (P<0.05), and the relative cerebral infarction areas(P<0.05),the water contents in brain tissue(P<0.05),the expression levels of CRP,the levels of TNF-α, IL-6, and IL-1β in brain tissue of the rats in nimodipine group and Shuxuening Injection group were significantly decreased(P<0.05), while the expression levels of GDF-15 were significantly increased(P<0.05).Compared with nimodipin group, the score of neurological impairment,the relative cerebral infarction area, the water content in brain tissue,the expression levels of CRP and GDF-15 and the levels of TNF-α, IL-6 and IL-1β in brain tissue of the rats in Shuxuening Injection group had no statistical significances (P>0.05). Conclusion: Shuxuening Injection can protect the acute cerebral infarction by up-regulating the expression of GDF-15 and inhibiting the expression of CRP in brain tissue of the rats with acute cerebral infarction, reducing the levels of inflammatory cytokines and improving the neurological function.

Objective: To observe the improvement effect of panax notoginseng saponins on the cardiac function of the rats with chronic heart failure(CHF), and to investigate the possible mechanism. Methods: Abdominal aortic constriction was used to establish the CHF rat models. Sixty model rats were randomly divided into model group, positive control group, low, medium and high doses of panax notoginseng saponins groups (n=12).Another 12 rats were taken as sham operation group. Color doppler ultrasound was used to detect the left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), left ventricular posterior wall diastolic thickness (LVPWD), left ventricular ejection fraction (LVEF), cardiac output (CO), left ventricular end-diastolic pressure (LVEDP), left ventricular systolic pressure (LVSP), maximal rate of increase of left ventricular pressure(+dp/dt max) and maximal rate of decrease of left ventricular pressure(-dp/dt max) of the rats in various groups,HE staining was used to observe the pathomorphology of myocardium tissue of the rats in various groups,TUNEL method was used to detect the apoptotic rates of cadiomyocytes of the rats in various groups,and Western blotting method was used to detect the expression levels of extracellular signal-regulated kinase(ERK),phosphorylation ERK (p-ERK),c-Jun NH2-terminal protein kinase(JNK),phosphorylation JNK(p-JNK),p38, and p-p38 proteins in myocardium tissue of the rats in various groups. ELISA was used to determine the serum levels of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) of the rats in various groups. Results: Compared with sham operation group, the LVEDD, LVESD, LVPWD, LVEDP and -dp/dt max of the rats in model group were significantly increased(P<0.05), while the LVEF, CO, LVSP, and +dp/dt max were significantly decreased(P<0.05). Compared with model group, the LVEDD, LVESD, LVPWD, LVEDP and -dp/dt max of the rats in positive control group and low, medium and high doses of panax notoginseng saponins groups were significantly decreased(P<0.05), while the LVEF, CO, LVSP and +dp/dt max were significantly increased (P<0.05).The cardiomyocytes of the rats in model group were hypertrophy and necrosis, and the myocardial fibers were irregular arrangement, with inflammatory cell infiltration and myocardial cell apoptosis. The structure of myocardium tissue of the rats in positive control group and low, medium and high doses of panax notoginseng saponins groups were nearly complete, the cardiomyocyte hypertrophy was relieved, and the myocardial fibers were slagking. Compared with sham operation group, the apoptotic rate of cardiomyocytes of the rats in model group was significantly decreased(P<0.05), the expression levels of p-ERK,p-JNK,and p-p38 proteins in myocardium tissue of the rats in model group were significantly increased(P<0.05), the serum levels of TNF-α and IL-6 of the rats in model group were significantly increased (P<0.05).Compared with model group, the apoptotic rates of cardiomyocytes of the rats in positive control group and low, medium and high doses of panax notoginseng saponins groups were significantly decreased(P<0.05), the expression levels of p-ERK,p-JNK,p-p38 proteins in myocardial tissue were significantly decreased (P<0.05), and the serum levels of TNF-α and IL-6 were significantly decreased(P<0.05). Conclusion: Panax notoginseng saponins can inhibit the apoptosis of the cardiomyocytes and improve the CHF by down-regulating the expression levels of p-ERK,p-JNK and p-p38 proteins and inhibiting the secretion of serum inflammatory factors.

Objective: To investigate the protective effect of emodin on the focal cerebral ischemia in the rats, and to clarify its mechanism. Methods: Sixty healthy male SD rats were randomly divided into sham operation group(given saline), model group(given saline,modeling),positive control group(give 5 mg·kg^-1 dexamethasone, modeling), low, medium and high doses of emodin groups(given 10,20,and 40 mg·kg^-1 emodin,modeling)(n=10). The middle cerebral artery occlusion(MCAO) was used to establish the rat models of focal cerebral ischemia.The behavioral scores of the rats in various groups were detected,the percentages of cerebral infarction volumes of the rats in various groups were determined by TTC staining,the levels of tumor necrosis factor (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6(IL-6) in hippocampus tissue at ischemic side of the rats in various groups were measured by enzyme linked immunosorbent assay(ELISA) method,and the expression levels of nuclear transcription factor-κB (NF-κB) p65 protein and nuclear transcription factor κB inhibitor-α(IκBα) protein in hippocampus tissue at ischemic side of the rats in various groups were detected by Western blotting method. Results: No neurological defects or cerebral infarction of the rats in sham operation group was found. Compared with sham operation group, the levels of IL-6,IL-1β, and TNF-α in hippocampus tissue of the rats in model group were significantly increased (P<0.05), while the expression level of NF-κB p65 protein was significantly increased (P<0.05), and the expression level of IκBα protein was decreased significantly (P<0.05).Compared with model group, the behavioral scores of the rats in positive control group, medium and high doses of emodin groups were significantly decreased(P<0.05),the percentages of cerebral infarction volumes were decreased(P<0.05), the levels of IL-6,IL-1β,and TNF-α in hippocampus tissue at ischemic side of the rats were significantly decreased(P<0.05), the expression levels of NF-κB p65 protein were significantly decreased(P<0.05),and the expression levels of IκBα protein were significantly increased(P<0.05).Compared with low dose of emodin group, the behavioral scores and the percentages of cerebral infarction volumes of the rats in medium and high doses of emodin groups were significantly decreased (P<0.05), the levels of IL-6,IL-1β,and TNF-α in hippocampus tissue at ischemic side of the rats were significantly decreased (P<0.05), the expression levels of NF-κB p65 protein were significantly decreased(P<0.05), and the expression levels of IκBα protein were significantly increased(P<0.05). Conclusion: Emodin has the protective effect on the focal cerebral ischemia, and its mechanism may be related to the inhibition of NF-κB signaling pathway and the secretion of inflammatory factors.

Objective: To investigate the effects of antimicrobial peptide LL-37 on the tumor growth and apoptosis of the mice with colon cancer, and to elucidate the possible molecular mechanism of its anti-tumor effect. Methods: The LL-37 over-expression colon cancer HT-29 cells were constructed, and the expression levels of LL-37 mRNA and protein in the HT-29 cells were detected by qRT-PCR and Western blotting methods. A total of 30 BALB/c mice were randomly divided into control group (given the uninfected HT-29 cells), empty vector group (given the HT-29 cells infected with empty plasmid), LL-37 over-expression group (given the HT-29 cells infected with LL-37 over-expression vector), AMPK inhibitor group[given the HT-29 cells infected with empty vector, and then injected with 2 mg·kg^-1 Dorsomorphin (Dor) in the tail vein] and combination group (given the HT-29 cells infected with LL-37 over-expression vector, and then injected with 2 mg·kg^-1 Dor in the tail vein),and there were 6 mice in each group. The colon cancer HT-29 cells were used to replicate the colon cancer models of the mice. The tumor volumes of the mice in various groups were detected and the growth curve was drawn. After infected for 24 d, the weights of the tumor of the mice in various groups were measured and the inhibitory rates of the tumor were calculated. TUNEL staining was used to detect the apoptosis of tumor cells in various groups. Western blotting method was used to detect the expression levels of autophagy-related proteins Beclin1, ATG5, LC3Ⅰ, LC3Ⅱ and AMPK/mTOR pathway-related proteins AMPK, p-AMPK, mTOR and p-mTOR in tumor tissue of the mice in various groups. Results: Compared with control group and empty vector group, the expression levels of LL-37 mRNA and protein in the HT-29 cells of the mice in over-expression group were significantly increased(P<0.01),the weight and volume of the tumor were significantly decreased (P<0.05), the inhibitory rate of tumor was increased(P<0.05), the number of apoptotic cells was increased, the expression levels of autophage-related proteins Beclin1 and ATG5 in tumor tissue and the ratios of LC3Ⅱ/LC3Ⅰ and p-AMPK/AMPK were increased(P<0.05), while the ratio of p-mTOR/mTOR was significantly decreased (P<0.05).Compared with empty vector group,the weight and volume of the tumor of the mice in AMPK inhibitor group were increased (P<0.05), the inhibitory rate of the tumor was decreased (P<0.05), the number of apoptotic cells was decreased, the expression levels of autophagy-related proteins Beclin1 and Atg5,the ratios of LC3Ⅱ/LC3Ⅰ and p-AMPK/AMPK were decreased (P<0.05), and the ratio of p-mTOR/mTOR was increased (P<0.05).Compared with LL-37 over-expression group, the expression levels of Beclin1, ATG5,the ratios of LC3Ⅱ/LC3Ⅰ and p-AMPK/AMPK in tumor tissue of the mice in combination group were significantly decreased (P< 0.05 or P<0.01), and the ratio of p-mTOR/mTOR was significantly increased (P<0.01). Conclusion: The antimicrobial peptide LL-37 can activate the AMPK/mTOR signaling pathway and promote the autophagy of the cells, thereby inhibit the tumor growth in the colon cancer HT-29-bearing mice.

Objective: To investigate the percentages of lymphocyte subsets, the transformation and proliferation activities of T lymphocytes, the expression levels of serum immunoglobulin and cytokines in the immunosuppressive model rats after given Bifidobacterium lactis M8, and to elucidate the effect of Bifidobacterium lactis M8 on the immune function of the immunosuppressive model rats. Methods: Fifty Wistar rats were randomly divided into blank control group, model control group, low dose of Bifidobacterium lactis M8 group, middle dose of Bifidobacterium lactis M8 group, and high dose of Bifidobacterium lactis M8 group, and there were 10 rats in each group.The rats in each group were administrated with probiotic bifidobacterium lactis M8 for 6 d.From the 7th day, except for blank control group, the rats in the other groups were given cyclophosphamide (CTX) for 3 d to construct the immunosuppressive models; the rats in low, middle, and high doses of Bifidobacterium lactis M8 groups were given 0.01, 0.10 and 1.00 g bifidobacterium lactis M8, and the rats in blank control group and model control group were given sterile saline with the same volume. The percentages of lymphocyte subsets (CD3+ T lymphocytes, CD4+ T lymphocytes, CD8+ T lymphocytes, B lymphocytes and NK cells) in whole blood of the rats in various groups were detected by flow cytometry, the transformation and proliferation activities of T lymphocytes of the rats in various groups were detected by CCK-8 method, the levels of serum immunoglobulin A (IGA), immunoglobulin G (IgG), interleukin-2(IL-2), interleukin-6(IL-6), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) of the rats in various groups were detected by ELISA method. Results: Compared with blank control group, the percentages of CD3+ T lymphocytes, CD4+ T lymphocytes, CD8+ T lymphocytes, B lymphocytes and NK cells in whole blood of the rats inmodel control group were decreased significantly (P<0.05); compared with model control group, the percentages of CD3+ T lymphocytes, CD4+ T lymphocytes and CD8+ T lymphocytes in whole blood of the rats in high dose of Bifidobacterium lactis M8 group were increased significantly (P<0.05 or P<0.01),and the percentages of B lymphocytes and NK cells were increased significantly (P<0.01).Compared with blank control group, the transformation and proliferation activities of T lymphocytes of the rats in model control group was decreased (P<0.01); compared with model control group, the transformation and proliferation activities of T lymphocytes of the rats in middle and high doses of Bifidobacterium lactis M8 group were increased (P<0.01). Compared with blank control group, the levels of serum IgA, IgG, IL-2, IL-6, IFN-γ and TNF -α of the rats in model control group were decreased (P<0.01); compared with model control group, the levels of serum IgA, IgG, IL-2, IL-6, IFN-γ and TNF-α of the rats in middle and high doses of Bifidobacterium lactis M8 group were increased (P<0.05 or P<0.01). Conclusion: Perfusion of Bifidobacterium lactis M8 can improve the specific and non-specific immune response regulation ability of the immunosuppressed model rats. Bifidobacterium lactis M8 has a promotion effect on the immune regulation of the immunosuppressed rats.

Objective: To investigate the expressions of heterogeneous nuclear ribonucleoprotein D (hnRNPD) and epithelial cell adhesion molecule (EpCAM) in the oral squamous cell carcinoma (OSCC) tissue and normal oral mucosa tissue, and to discuss the associations between their expressions and the clinicopathological parameters of the OSCC patients,and to elucidate their effects on the occurrence and development of OSCC. Methods: The tumor tissue of 38 OSCC patients without preoperative radiotherapy and chemotherapy(OSCC group)and the normal oral mucosa tissue of 11 patients with tooth extraction (control group) were selected. Immunohistochemistry staining was used to detect the expression levels of hnRNPD and EpCAM in the tumor tissue and normal oral mucosa tissue,the expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients with different clinicopathological parameters were analyzed,and the correlations between the expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients were detected by Spearman correlation test. Results: Compared with control group, the expression levels of hnRNPD and EpCAM in the tumor tissue in OSCC group were increased (Z=-4.936, P=0.000; Z=-2.780, P=0.005); there was statistically significant difference in the expression level of EpCAM in tumor tissue of the OSCC patients with different genders and ages (Z=-2.471, P=0.013;Z=-1.967; P=0.049), there was no significant difference in the expression level of hnRNPD in tumor tissue of the OSCC patients with different clinicopathological parameters(P>0.05). The correlation between the expression levels of hnRNPD and EpCAM was weak (ρ=0.227, P=0.17). Conclusion: The expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients are up-regulated, and they promote the occurrence and development of OSCC.

Objective: To measure the morphological parameters of the radial head of the Chinese and compare with those of the Western population, and to provide the references for the design of the radial head prosthesis of the Chinese. Methods: The CT scan data of normal elbow from 130 Chinese cases were selected. All the data were scanned by using a 64-slice spiral CT scanner and reconstructed and measured on AW 4.5 workstations. The angle between neck and shaft (∠α),the proximal radial length of neck and head (LNH), the maximum diameter of radial head (D1MAX), the minimum diameter of radial head (D2MIN), the maximum diameter of articular surface (D3MAX) and the maximum vertical depression depth (DD3),the angle between D3MAX and horizontal plane (∠β3),the minimum diameter of articular surface (D4MIN) and the minimum vertical depression depth (DD4), and the angle between D4MIN and horizontal plane (∠β4) were measured. The measurement results of the male and female were compared and the correlations between different parameters were analyzed. Results: The measured values of ∠α, LNH, D1MAX, D2MIN, D3MAX, DD3, D4MIN and DD4 of the normal male were larger than those of the normal female (P<0.01). There were no significant differences in the measured values of ∠β3 and ∠β4 and the ratios of DD3/D3MAX, D4MIN/D3MAX, ∠β3/∠β4 and D2MIN/D1MAX between the normal male and the normal female (P>0.05). There was a strong positive correlation between ∠α, LNH, D1MAX, D2MIN, D3MAX and D4MIN (P<0.05), and there was also a strong positive correlation between ∠β3 and ∠β4(P<0.05). Conclusion: The morphology of radial head of the normal male Chinese is similar to the female. The morphological parameters of the radial head in the Chinese people are different from those in other countries.

Objective: To observe the expression of valosin-containing protein(VCP) in the epithelial ovarian cancer (EOC) tissue and its relationships with the clinicopathological features of the EOC patients,and to provide the basis for the molecular treatment of EOC. Methods: The expressions of VCP in 94 EOC tissue samples,13 ovarian borderline tumor tissue samples,36 ovarian benign tumor tissue samples and 8 normal ovarian tissue samples were measured by immunohistochemical method. The relationships between the VCP expression and the clinicopathological parameters (age,FIGO stage,pathological type,histological grade, lymph node metastasis or not, ascites or not, preoperative CA125 level) of the EOC patients were analyzed.The expressions of VCP in human ovarian epithelial HOSEPIC cells and human EOC SKOV-3 cells were detected by immunofluorescence technique and Western blotting method.The SKOV-3 cells were divided into control group(without CB-5083) and treatment group (with CB-5083), the migration rates of the SKOV-3 cells in two groups were analyzed by scratch experiment, and the clone formation rates of the SKOV-3 cells in two groups were analyzed by plate clone formation experiment. The expressions of VCP in the cells in two groups were analyzed by immunofluorescence technique.The expression levels of VCP, NF-κB/P65,IκBα,and p-IκBα proteins in the SKOV-3 cells in two groups were detected by Western blotting method. Results: The positive expression rates of VCP in EOC, borderline ovarian tumor, benign ovarian tumor and normal ovarian tissues had significant difference (P<0.05);the expression of VCP was associated with the FIGO staging, histological grade, and lymph node metastasis or not(P<0.05), but was not associated with the age, pathological type, ascites or not, and preoperative Ca125 level of the EOC patients (P>0.05).The VCP expression level in the SKOV-3 cells was higher than that in HOSEpiC cells(P<0.05).The migration rate of the SKOV-3 cells in treatment group was lower than that in control group (P<0.05),the cloning formation rate of the SKOV-3 cells in treatment group was lower than that in control group(P<0.05).The expression levels of VCP and NF-κB/P65 proteins of the SKOV-3 cells in treatment group were lower than those in control group (P<0.05),and the expression level of p-IκBα protein was higher than that in control group (P<0.05). Conclusion: VCP is highly expressed in the EOC tissue and cells, and high-expression VCP can promot the tumor migration and proliferation.

Objective: To detect the expression levels of miR-194-5p and magnesium transporter protein1(MagT1) in peripheral blood mononuclear cells (PBMCs) of the patients with chronic hepatitis B virus(HBV) infection, and to investigate their possible clinical significances. Methods: The clinical materials of 40 healthy subjects (healthy control group), 40 cases of asymptomatic hepatitis B carriers(HBV carrier group), 40 patients with mild-moderate chronic hepatitis B (mild-moderate CHB group) and 40 patients with severe chronic hepatitis B (severe CHB group) were collected. The peripheral blood of the subjects in various groups was collected and the PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation. The expression levels of miR-194-5p and MagT1 mRNA in PBMCs of the subjects in various groups were detected by RT-PCR assay, the expression levels of MagT1 protein in PBMCs of the subjects in various groups were detected by Western blotting method,the levels of Mg²⁺ in peripheral blood and PBMCs of the subjects in various groups were detected, and the expression levels of CD8+ T lymphocyte surface molecules programmed death receptor 1(PD-1) and natural killer cell receptor 2 group D molecule (NKG2D) in PBMCs of the subjects in various groups were analyzed by flow cytometry. The miR-194-5p mimic (miR-194-5p mimic group)and miR-NC(miR-NC group) were transfected into the 293T cells, and then the dual luciferase reporter gene assay was used to detect the luciferase activities in 293T cells in two groups. Results: Compared with healthy control group, the expression levels of MagT1 mRNA and protein, the expression levels of CD8+ T lymphocyte surface molecule NKG2D and the levels of Mg²⁺ in PBMCs of the patients in HBV carrier group, mild-moderate CHB group and severe CHB group were decreased(P<0.05),the expression levels of miR-194-5p mRNA in PBMCs and CD8+ T lymphocyte surface molecule PD-1 of the patients were increased(P<0.05).Compared with HBV carrier group and mild-moderate CHB group,the expression levels of MagT1 mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg²⁺ in PBMCs of the patients in severe CHB group were decreased (P<0.05), and the expression levels of miR-194-5p mRNA in PBMCs and CD8+ T lymphocyte surface molecule PD-1 of the patients were increased (P<0.05).Compared with HBV carrier group, the expression levels of MagT1 mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg²⁺ in PBMCs of the patients in mild-moderate CHB group were decreased(P<0.05),and the expression levels of miR-194-5p mRNA in PBMCs and CD8+ T lymphocyte surface molecule PD-1 of the patients were increased (P<0.05).The dual luciferase reporter gene assay results confirmed that MagT1 was a miR-194-5p target gene. Conclusion: MiR-194-5p may lead to chronic development of hepatitis B by down-regulating the expression of MagT1, causing MagT1-mediated Mg²⁺ influx disorder and resulting in the decrease of the immune activity of CD8+ T lymphocyte.

Objective: To detect the levels of B7-H4 protein, adenosine deaminase (ADA) and carcinoembryonic antigen (CEA) in the malignant pleural effusion associated with lung cancer (LC-MPE) and tuberculous pleural effusion (TPE), and to evaluate the values of single and combined detection of B7-H4 protein,ADA and CEA in differential diagnosis of LC-MPE and TPE. Methods: A total of 202 samples of pleural effusion(PE) from 120 LC-MPE patients(LC-MPE group) and 82 TPE patients(TPE group) were collected. The levels of ADA and CEA and B7-H4 protein in PE were detected by the enzymatic, electrochemiluminescence and ELISA methods, respectively; and the receiver operating characteristic curve(ROC curve) was used to determine the diagnostic efficiencies of the above indexes alone or in combination,such as the sensitivity,the specificity, the Yoden index(YI),and the area under ROC curve(AUC). Results: The level of ADA in PE of the patients in TPE group was higher than that in LC-MPE group (P<0.05); the levels of CEA and B7-H4 protein in PE of the patients in LC-MPE group were higher than those in TPE group (P<0.05).The ROC curve analysis results showed that the sensitivities of single detection of ADA, CEA and B7-H4 protein were 93.30%, 83.33% and 79.90%, respectively; the specificities were 86.59%, 96.34% and 72.50%, respectively; the AUC were 0.927, 0.925 and 0.836, respectively. The combined detection of the three indexes had the highest diagnostic value; the sensitivity was 93.90%, the specificity was 97.50%, the YI was 0.971, and the AUC was 0.998. Conclusion: The combined detection of levels of ADA, CEA and B7-H4 protein in PE is superior to the single evaluation of each index, which can greatly improve the differential diagnosis efficiencies of LC-MPE and TPE.

Objective: To analyze the clinical features of the patient with nephrotic syndrome who developed pneumocystis carinii pneumonia(PCP)and cytomegalovirus pneumonia(CMP) after oral administration of tacrolimus caspsules,and to discuss the correlations between immunosuppressive patient and pneumocystis carinii(Pc) and cytomegalovirus(CMV) infection, and to provide the basis for the reasonable treatment plan in the early stage. Methods: The clinical materials of one patient with nephrotic syndrome who developed PCP and CMP after oral administration of tacrolimus capsules were collected and the clinical symptoms, past medical history and outcomes, auxiliary examination, treatment plan and prognosis were analyzed; the relevant literatures were reviewed. Results: The male 47-year-old patient was admitted to hospital because of cough for 1 month, shortness of breath for 1 week and fever for 3 d. The patient had the history of diabetes mellitus and took the medication regularly, and the level of blood sugar was well controlled. At the beginning of 2018, the patient received renal biopsy due to edema of the lower extremities and was diagnosed as stage Ⅱ membranous nephropathy accompanying with mild mesangial proliferative diabetic nephropathy; the patient was orally administrated with glucocorticoid. In July 2018, the patient was diagnosed as nephrotic syndrome and stage Ⅱ membranous nephropathy,and had been orally administrated with tacrolimus capsules after discharge. After admission, the patient developed acute respiratory distress syndrome rapidly;the multiple exudation and nodular foci of both lungs were found on the chest imaging, and the infectious lesions were considered. The IgM antibody and IgG antibody of CMV of the patient were both positive.The high throughput gene detection results of the infection pathogens in blood showed Pneumocystis jiroveci of Pneumocystis and human herpesvirus 5(HHV-5).PCP complicated with CMP was diagnosed definitively. The patient was treated with sulfamethoxazole combined with ganciclovir and noninvasive ventilation.The patient was discharged after the condition was improved. Conclusion: The patient with low immunity should be alert to the mixed infection of PCP and CMP if he develops rapidly progressive hypoxemia.

Objective: To investigate the effect of modified fecal drainage device on avoiding the occupational exposure infection of the medical staffs in the treatment of corona virus disease 2019(COVID-19), and to provide the reference for the clinical treatment of COVID-19 and avoiding the occupational exposure infection of the medical staffs. Methods: The clinical data of a critical COVID-19 patient with diarrhea as the main symptom were collected.The modified fecal drainage device of F18 silicone gastric tube connected with disposable negative pressure drainage device was uesd to treat the fecal excrement of the patient.The general data of the medical staffs,contaning 16 doctors and 48 nurses,were collected.The COVID-19 serological antibodies and pharyngeal swabs of the medical staffs were tested every 2 weeks. Results: The 78-year-old woman patient was admitted to hospital due to diarrhea, cough and expectoration for 15 d, chest distress and shortness of breath for 10 d,and fever for 1 d.The test result of COVID-19 pharyngeal swabs of the patient was positive.After the feces were collected with the modified fecal drainage device, the average operation time of medical staffs was reduced from 20 min to 10 min, the patient's perianal skin flushing subsided, and no incontinence-associated dermatitis(IAD)occurred. The patient was cured but remained in hospital for the other underlying diseases. The test results of COVID-19 serological antibodies and pharyngeal swabs of 64 medical staffs were all negative, all the medical staffs had no infection. Conclusion: The modified fecal drainage device has better stability, which can effectively prevent IAD and the spread of COVID-19 and reduce the risk of occupational exposure infection of medical staffs, and it is suitable for clinical promotion application.

Objective: To analyze the clinical features, imaging findings, diagnostic methods, pathomorphology and prognosis of the patients with primary pulmonary diffused large B-cell lymphoma (DLBCL) with the multiple ground glassy nodule shadows in the lungs as the main manifestation, and to improve the clinicians'understanding of the primary pulmonary DLBCL. Methods: The clinical data of a patient with primary pulmonary DLBCL were collected. The main manifestation in CT examination of the patient was multiple massive ground glass nodule shadows in the lungs. The initial diagnosis was pneumonia. The final diagnosis of primary pulmonary DLBCL was confirmed by pathology and immunohistochemistry examination. The related literatures were reviewed. Results: The female patient was admitted to hospital due to cough and expectoration as the first manifestations. There were no obvious positive signs in the physical examination. The CT examination results showed multiple ground glassy nodular shadows in both lungs.The tumor markers, lymph node color Doppler ultrasound, PET-CT, bone marrow biopsy, pathological biopsy and the other related examinations were performed, and the related treatment were given. The PET-CT results showed the multiple ground glass nodule in both lungs complicated with increasing of partial metabolism, and the pathological and immunohistochemical results suggested that the patient was non-Hodgkin DLBCL originated from the activated B cells outside the germinal center,and the final diagnosis was primary pulmonary DLBCL. The patient was given R-CHOP regimen regularly.The patient received chest CT examination 3 months later,and the results showed that the massive nodule shadows in both lungs were disappeared. Conclusion: The clinical features and imaging findings of the patient with primary pulmonary DLBCL are not specific, its diagnosis ultimately relies on the pathomophological and immunohistochemical examination results; the degree of malignancy of primary pulmonary DLBCL is high,and the prognosis is poor; when there are multiple ground glass nodule shadows in the lungs, the disease should be taken into account.

Objective: To investigate the clinical characteristics and diagnosis and treatment process of a patient with invasive mucinous adenocarcinoma (IMAs)of lung,and to improve the clinician's understanding of IMAs. Methods: The general materials, imaging manifestations and treatment plan of a patient with IMAs were collected, and the related literature review was conducted. Results: A 42-year-old female patient was admitted to hospital due to cough and expectoration, the CT examination results showed the bilateral lung patchy shadows, the patient was suspected of having pneumonia.After anti-infective treatment, the patient's symptoms did not improve. The pathological findings of transbronchial lung biopsy(TBLB) and the examination of exfoliated cells of pleural fluid all showed inflammation, and the pathological result of percutaneous biopsy was IMAs. The result of gene detection was 2-point mutation in exon of KRAS. After chemotherapy with paclitaxel plus carboplatin combined with bevacizumab, the symptoms of the patients were improved significantly, and the changes of imaging manifestations were obvious. Conclusion: IMAs is a special pathological type of lung adenocarcinomas (ADCs) with various imaging manifestations and specific gene expression. The treatment principles are different from those of the other types of ADCs.

Objective: To analyze the clinical manifestations, pathogenesis, auxiliary examinations, differential diagnosis and treatment methods of the patient with synovial chondromatosis of temporomandibular joint (TMJSC)with disc perforation,and to provide the basis for the diagnosis and treatment of TMJSC patients. Methods: The clinical data of one patient with TMJSC were collected,and the diagnosis and treatment process was analyzed, and the related literatures were reviewed. Results: The female 53-year-old patient was admitted to hospital due to occasional tenderness in the right TMJ for more than 1 year. The computerized tomography (CT)results revealed that the changes could be found around the wall of maxillary sinus and the joint. The magnetic resonance imaging(MRI) results showed the diffused nodular, low and equal signals in the joint cavity, the shadow of fluid signals in the joint cavity was increased, and the joint space was broadened.The open surgery was performed in the patient successfully, including the removal of loose bodies and synovectomy, and the pathology result after opration was synovial chondromatosis(SC). No recurrence was found during 6 months after follow-up. Conclusion: The histopathology examination assisted with CT and MRI should be perfomed in the diagnosis of TMJSC. Surgical treatment is an effective method,and the pathological diagnosis is the gold standard for TMJSC.