Objective:To investigate the effect of co-expression of sleeping beauty(SB) transposon and IL-10 mediated by adenovirus on the balance of Th17/Treg cells in the splenocytes of the non-obese diabetes(NOD) mice,and to illuminate the possible mechanism of IL-10 in the treatment of NOD mice. Methods: The splenocytes were extracted from spleen of the C57BL6 mice and cultured.The splenocytes were divided into control group,empty vector group and therapy group.The cells in control group didn't receive any treatment,the cells in empty vector group were co-infected with the adenovirus vector without IL-10 gene containing transposon sequence and the adenovirus vector without transposase,and the cells in therapy cells were co-infected with the adenovirus vector containing IL-10 gene and transposon sequence and the adenovirus vector with transposase.After 48 h of infection,the expression leves of IL-10 mRNA in the splenocytes of the mice in various groups were assessed by RT-PCR.The cells of above three groups were subcutaneously injected into popliteal space in right hind leg of the NOD mice in control group,empty vector group and therapy group;there were six mice in each group;once a week and six times.The mice were killed 4 weeks after injection,the expression levels of IL-10 in serum of the NOD mice in various groups were assessed by ELISA,and the proportions of CD4+IL-10+,CD4+IFN-γ+,Th17 and Treg cells in the splenocytes were detected by flow cytometry. Results: Compared with control group and empty vector group,the expression level of IL-10 mRNA in splenocytes of the C57BL6 mice in therapy group was increased(F=72.71,P<0.05); the expression level of IL-10 in serum of the NOD mice was increased(F=8.89,P<0.05); the proportions of CD4+IL-10+ cells and Treg cells were significantly increased (F=72.09,P<0.05;F=12.98,P<0.05); the proportion of Th17 cells was decreased(F=6.39,P<0.05).The proportion of CD4+IFN-γ+ cells had no significant differences between various groups (F=2.72,P>0.05). Conclusion: The co-expression of SB transposon and IL-10 can significantly increase the IL-10 level in serum of the NOD mice,increase the proportion of CD4+IL-10+ cells,decrease the proportion of Th17 cells and increase the proportion of Treg cells in the splenocytes,which can regulate the balance of Th1/Th2 and Th17/Treg cells and play a therapeutic effect in the NOD mice.

Objective:To investigate the effect of Lyn on the expression of MUC5AC in human bronchial epithelial cells induced by house dust mite(HDM),and to explore its possible mechanism. Methods: The human bronchial epithelial cells(16HBE) were divided into PBS group and HDM group (1μg·L^-1 HDM).The cells were transfected by liposome.The luciferase report gene of MUC5AC promoter was constructed.The relative luciferase unit(RLU) was detected by double luciferase report gene assay.The expression of MUC5AC in the cells was detected by immunofluorescence technique and observed by confocal microscope.The expression level of STAT6 in the 16HBE cells was detected by Western blotting method. Results: The results of double luciferase report gene assay showed that the RLU in HDM group was higher than that in PBS group(P<0.05).The RLU of cells in HDM group treated with LynsiRNA intervention was higher than that of the cells without LynsiRNA intervention(P<0.05).The immunofluorescence results demonstrated that the expression level of MUC5AC in HDM group was higher than that in PBS group(P<0.05),and the expression level of MUC5AC in HDM group was increased after LynsiRNA intervention(P<0.05).The Western blotting results indicated that the expression level of STAT6 was up-regulated in HDM group when the cells were intervened with LynsiRNA(P<0.05). Conclusion: Deficiency of Lyn can increase the expression of MUC5AC in the human bronchial epithelial cells,and its mechanism may be related to regulating the STAT6 signal pathway by Lyn.

Objective:To explore the protective effect of exosomes secreted from bone mesenchymal stem cells(BMSCs) modified by hypoxia inducible factor-1α(HIF-1α) on the chondrocytes,and to elucidate the possible mechanism of its combination with cartilage regenerated scaffolds in promoting the repair of advanced cartilage defects. Methods: The exosomes(BMSCs-Exo^WT and BMSCs-Exo^MU) were extracted from the BMSCs modified by wild type of HIF-1α and mutant type of HIF-1α by ultracentrifugation method and identified in the meantime.In vitro the inflammatory response of chondrocytes were induced by interleukin-1β(IL-1β),the same amount of PBS,BMSCs-Exo^WT(80 μg·mL^-1),BMSCs-Exo^MU(80 μg·mL^-1) were respectively cultivated with the chondrocytes under the inflammatory reaction and blank group,inflammation group,BMSCs-Exo^WT group and BMSCs-Exo^MU group were set up;Hoechst33342 staining was used to detect the number of apoptotic bodies of chondrocytes in various groups.The Western blotting method was used to detect the expression levels of AKT/p-AKT,ERK/p-ERK and p38/p-p38 in the chondrocytes in various groups.Twelve New Zealand white rabbits were randomly divided into 4 groups and the models of rabbit knee cartilage defects were consructed; the equal volume of physiological saline,scaffold +physiological saline,scaffold +BMSCs-Exo^WT and scaffold +BMSCs-Exo^MU were respectively injected into the cartilage defects of rabbits.Six weeks after operation,gross conference,HE and safranin O staining were used to observe and compare the repair effects of cartilage defects in each group. Results: BMSCs-Exo^WT and BMSCs-Exo^MU were successfully extracted and identified,and the exosomes were observed to be nearly circular with diameter of about 40-100 nm;the Western blotting results showed that they expressed special proteins CD63 and CD81, respectively.In vitro,the number of apoptotic bodies of chondrocytes in BMSCs-Exo^MU group was lower than those in inflammation group and BMSCs-Exo^WT group(P<0.01).The Western blotting results revealed that the expression levels of p-ERK1/2 in BMSCs-Exo^MU and BMSCs-Exo^WT groups were lower than that in inflammation group(P<0.05); the expression levels of p-AKT and p-p38 were higher(P<0.05); the effect in BMSCs-Exo^MU group was stronger than BMSCs-Exo^WT group,and the difference was statistically significant(P<0.05).In the advanced cartilage defect models of rabbit knee joint,the repair effect in scaffold+ BMSCs-Exo^MU group was better than those in blank group,scaffold group and scaffold+BMSCs-Exo^WT group. Conclusion: Cartilage scaffold combined with BMSCs-Exo^MU can promote the repair of cartilage defects.

Objective:To evaluate the anti-adhesion properties of xyloglucan(mXG)hydrogel in the L929 mouse fibroblasts,to establish the animal model of recurrent adhesion in the rats after adhesionlysis,and to investigate the prevention effect of mXG hydrogel in recurrent adhesion and its influence in the expressions of transforming growth factor-β1(TGF-β1) and connective tissue growth factor (CTGF). Methods: After seeding on the surface of mXG gel,the adhesion property of L929 mouse fibroblasts was detected with fluorescence staining.The rat models of recurrent adhesion were established after adhesionlysis.Fourty-eight SD rats were randomly divided into 3 groups respectively(n=16).In adhesion group,1 mL saline was injected into the abdominal wall and cecum of the rats; in commercial membrane group,the 2 cm×3 cm chitosan anti-adhesion membrane was used to cover the wound of abdominal wall of the rats;in mXG hydrogel group,1 mL 4% mXG hydrogel was injected into the abdominal wall and cecal wound of the rats,and abdominal surgery was ended after the complete formation of the hydrogel (3 min).Eight rats were killed in each group 7 and 14 d after the second operation,and the degrees of adhesion were evaluated and the histological changes were observed.Immunohistochemical staining was used to observe the expression levels of CTGF and TGF-β1. Results: A large number of L929 mouse fibroblasts proliferated well and exhibited a spherical morphology in control group; but in mXG hydrogel group,only very little L929 mouse fibroblasts were globular,showing the mXG hydrogel had good resistance to the adhesion of L929 mouse fibroblasts.Blunt separate adhesion surface formed in all of the rats after operation.7 d after the second operation, 4-5 score adhesion with fibrous connective tissue hyperplasia formed in adhesion group;moderate adhesion formed in commercial membrane group with more connective tissue;most cecum and abdominal wall began to heal in mXG hydrogel group with less connective tissue.14 d after the second operation,more severe peritoneal adhesions presented in adhesion group with proliferated fiber connetive tissue and collegan;severe adhesions also presented in commercial membrane group; mild adhesion was found in two rats mXG group,the other rats had no adhesion, and the defects were almost completely recovered.On days 7 and 14 after the second operation,the mean adhesion score of the rats in mXG group was significantly lower than those in adhesion group and commercial membrane group(P<0.05).The expression levels of TGF-β1 and CTGF were increased with the increase of peritoneal adhesion scores and collagen deposition;the expression levels of TGF-β1 and CTGF were the highest in adgesion group and the lowest in mXG group. Conclusion: mXG hydrogels have good resistance to fibroblast adhesion,and mXG hydrogel is effective in reducing the formation of recurrent adhesion in the rats after adhesionlysis and can decrease the expression levels of adhesion-related factors TGF-β1 and CTGF.

Objective:To investigate the regulation effect of Wnt5a on the apoptosis of lung adenocarcinoma A549 cells,and to clarify its mechanism. Methods: The human lung adenocarcinoma cells were selected.The A549 cells treated with Wnt5a were used as treatment group,and the A549 cells treated with C culture solution were used as control group.The apoptotic body induced by Wnt5a was assessed with TUNEL assay; the apoptotic rates of the A549 cells in various groups were detected by Annexin Ⅴ-FITC/PI double staining;the reactive oxygen species(ROS) levels in the A549 cells in various groups were determined with DCFH-DA fluorescence probe,and the mitochondria membrane potential was assessed with JC-1 staining method.Western blotting was used to analyze the expression levels of apoptosis-related proteins in the A549 cells in various groups. Results: Compared with control group,the apoptotic rates of the A549 cells in treatment group 12,24,and 48 h after treatment were significantly increased(P<0.01);the ROS levels were increased(P<0.05);the mitochondria membrane potentials were decreased(P<0.05),the expressing amount of BAX was up-regulated and the expression amount of AIF was down-regulated. Conclusion: Wnt5a has regulation on the apoptosis of human lung adenocarcinoma cells and can promote the apoptosis of A549 cells through mitochondrial pathway.

Objective:To explore the feassibility to edit human p53 and PTEN, two tumor suppressor genes, by CRISPR-Cas9 technology and to evaluate the editing efficiency in vitro,and to provide the experimental study tools for transforming primary healthy cells into malignant tumor cells and establishment of humanized mouse models with human oncogenesis in vivo. Methods: The single-guide RNA (sgRNA) sequences were designed to target the common exon regions of p53 and PTEN mRNA isoforms based on software analysis, that could predict their gene editing efficiency. The sgRNAs with high scores were selected and cloned into Cas9-P2A-GFP plasmid to construct sgRNA-Cas9-P2A-GFP vector that co-expressed sgRNA, Cas9 and GFP. The 293T cells in logarithmic growth phase were transfected with the sgRNA-Cas9-P2A-GFP vector (experimental group) or PBS (control group) by Lipofectamine 2000. After two-week expansion, the GFP-positive 293T cells were purified by flow cytometric sorter, whose genomic DNA was extracted for further analysis. The DNA fragments containing the sgRNA targeting site were amplified from the extracted genomic DNA by PCR and purified by gel extraction. Then they were linked into the pEASY-Blunt Zero cloning vector and transformed into competent E. coli cells. The single colonies formed by pEASY-Blunt Zero vector transformed cells were used to extract the plasmid for DNA sequencing. And the sequencing results of control group and experimental group were compared to judge the gene editing efficiency. Results: Over 82% of the sgRNA-Cas9-P2A-GFP transfected cells were found to express GFP gene after flow sorting in experimental group, which was significantly higher than that of the pre-sorted cells (P<0.05). Genomic DNA was extracted from the sorted cells after expansion and used as PCR template. The length of the amplified fragments containing the p53 mutation site was 612 bp, while the lengths of the amplified fragments containing the PTEN-1/PTEN-2 mutation site were 667 and 947 bp. The results of sequencing showed that the efficiency of editing induced by p53-1, p53-2 and PTEN-2 sgRNA were 54.5%, 45.5% and 33.3%, respectively. Conclusion: The sgRNAs p53-1, p53-2 and PTEN-2 designed for p53 and PTEN can successfully guide Cas9-mediated site-specific genome editing with high efficiency at the genome level.

Objective:To investigate the application of CD44 receptor-targeted nanoparticles HA_CDDP-DOX in the treatment of breast cancer, and to clarify its inhibitory effect on allograft breast cancer in the mice. Methods: Hyaluronic acid (HA) was used to construct a breast cancer targeted nanoparticle via green synthesis approach, and its particle size, stability and doxorubicin release profile at different pH conditions were measured. Then the breast cancer models were constructed by inoculating 4T1 cells into the mouse mammary fat pad. The model mice were randomly divided into control group, DOX/CDDP group and HA_CDDP-DOX group according to the tumor volumes and the body weights. PBS, free drug DOX/CDDP and HA_CDDP-DOX were intravenously injected into the allograft breast cancer models on the 1st, 5th and 9th days after the tumor volume reached about 80 mm³. The antitumor effect and biosafety of HA_CDDP-DOX were evaluated by detecting the tumor volume, mouse weight and immunohistopathology. Moreover, the biological distribution of HA_CDDP-DOX in the mice was studied by biofluorescence imaging. Results: The average particle size of HA_CDDP-DOX was (80.0 ±17.4) nm, which was stable under pH 7.4. Under acidic condition, the particle size was increased and the DOX was effectively released. Compared with DOX/CDDP group, the body weight of the mice in HA_CDDP-DOX group was increased (P<0.05) and the tumor volume was decreased (P<0.05). The HE staining results showed that the liver of the mice in control group had tumor metastasis and the alveolar septum was widened. The tumor tissue of the mice in DOX/CDDP group and HA_CDDP-DOX group were all necrotic, while in HA_CDDP-DOX group the degree of necrosis was significantly higher than DOX/CDDP group. Compared with DOX/CDDP group, the activity of Caspase-3 in HA_CDDP-DOX group was significantly increased (P<0.01), while the Ki-67 activity was significantly decreased (P<0.01). The biofluorescence imaging showed that the nanoparticle HA_CDDP-DOX could accumulate to the tumor site by targeting, and effectively release the drug. Conclusion: HA_CDDP-DOX nanoparticles can effectively target the breast cancer cells, reduce the toxicity of chemotherapy drugs, and enhance the therapeutic effect of breast cancer.

Objective:To explore the effect of differentiation of cardiac stem cells(CSC) mediated by pilose antler polypeptides on the expressions of terminal myocardial differentiation genes atrial natriuretic polypeptide(ANP) and myosin light chain 2v(MLC-2v),and to clarify the mechanism of repairing the damaged myocardium. Methods: The healthy male Wistar rats born 2 d were selected to extract the CSC.The surface marker c-Kit and the purity of CSC were identified by immunofluorescence and flow cytometry.The culture dish was used as the experimental unit and the extracted cells were divided into the following four groups:blank control group (the same amount of buffer was added for induction),5-azacytidine group(induced with 3 μmol·L^-1 5-azacytidine),pilose antler polypeptides group(induced with 800 mg·L^-1 pilose antler polypeptides) and combined group(induced with 800 mg·L^-1 pilose antler polypeptides and 3 μmol·L^-1 5-azacytidine);the cells were incubated for 48 h in the condition of 37℃ and 5% CO₂.The expression levels of ANP and MLC-2v in supernatant of the cells in various groups were detected by ELISA method.The expression levels of ANP and MLC-2v mRNA in the cells in various groups were detected by RT-PCR method. Results: CSC were prepared with the purity>95%.The results of ELISA showed that the expression levels of ANP and MLC-2v in 5-azacytidine, pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05).The expression levels of ANP and MLC-2v in combined group were increased compared 5-azacytidine and pilose antler polypeptides groups,but there were no significant differences(P>0.05).The result of RT-PCR showed that the expression levels of ANP and MLC-2v mRNA in 5-azacytidine,pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05). Conclusion: Pilose antler polypeptides can induce the differentiation of CSC into cardiac cells by promoting the expressions of ANP and MLC-2v, and they can reduce the cytotoxicity induced by 5-azacytidine in some degrees.

Objective:To investigate the transport pathwayand intracellular distribution of the of fluorescent carbon dots(CDs) synthesized by folic acid and polyethyleneimine(PEI) through the membrane of MC3T3-E1 cells and its effect on the cells, and to clarify the mechanism. Methods: The fluorescent CDs with the function of cell imaging were synthesized by hydrothermal method using folic acid and PEI as the raw materials; MTT assay was applied to screen the best concentration of CDs. The MC3T3-E1 cells were divided into blank control group, folic acid group and CDs group. The biocompatibility of CDs was evaluated by the detection of cell cycle, apoptosis and cellular reactive oxygen species(ROS) level. Nystatin as a kind of caveolae inhibitor and nocodazole as a kind of macropinocytosis inhibitor were used to find out the pathway through which the cells took in the CDs. Using the charcteristic of CDs with blue fluorescence stimulated by ultraviolet ray,the organelle probes were used to observe the distribution of CDs. Results: Compared with blank control group, the cells in different concentrations(100-450 mg·L^-1) of CDs groups showed no cytotoxicity at 24 h (P>0.05);at 48 h, the cell proliferation rate was reduced to 68.4% of blank control group when the concentration of CDs reached 350 mg · L ^-1(P<0.05).Compared with blank control group, the percentages of cells in G₀ phase and G₁ phase in CDs group were decreased (P<0.05), and the percentage of cells in S phase was increased (P<0.05); the percentages of cells in G₂ phase and M phase were increased, but there no was significant differences (P>0.05). Compared with blank control group, the apoptotic rates of the cells in folic acid group and CDs group had no significant differences (P>0.05). Compared with blank control group,the intracellular ROS levels in folic acid group and CDs group were significantly decreased(P<0.05). Compared with blank control group, the uptake amount of CDs in the cells was decreased in nystatin group(P<0.05). The blue fluorescence of CDs overlapped with the red fluorescence of mitochondria under an inverted fluorescence microscope, the blue fluorescence of CDs overlapped with the red fluorescence of lysosomes;they didn't overlap completely with the red fluorescence of the endoplasmic reticulum;the blue fluorescence of CDs overlapped poorly with the red fluorescence of Golgi apparatus. Conclusion: CDs perform well in biocompatibility and they can be distributed to different organelles after taken in by the cells. They can be used as a kind of gene carrier in transgenic therapy.

Objective:To investigate the changes of PERK,Runx2,osterix,RANKL and OPG in bone tissue of the female rats with experimental postmenopausal osteoporosis (PMOP) before and after treatment,and to elucidate the role of PERK signaling pathway in PMOP. Methods: The ovariectomized rats were reproduced to osteoporosis models.A total of 45 rats were divided into normal control group (the rats didn't receive any treatment,n=15),osteoporosis group (the rats were ovariectomized,n=15) and osteoporosis treatment group (the ovariectomized rats were injected with estrogen through caudal vein,n=15).The changes of serum collagen Ⅰ (Col Ⅰ),alkaline phosphatase (ALP) and osteocalcin (OCN) of the rats in various groups were observed.Three months after feeding,the femoral shaft of the rats in various groups were taken for pathological section.The gene expression levels of PERK,ATF4,Runx2,osterix,RANKL and OPG in bone tissue of the rats in various groups were detected by RT-PCR; the protein expression levels of PERK,ATF4,Runx2,osterix,RANKL and OPG were detected by Western blotting method. Results: Compared with control group,the levels of serum Col Ⅰ,ALP and OCN in the rats in osteoporosis group were significantly decreased (P<0.05 or P<0.01); compared with osteoporosis group,the levels of serum Col Ⅰ,ALP and OCN of the rats in osteoporosis treatment group were significantly increased (P<0.01).Compared with control group,the gene expression levels of PERK,ATF4,Runx2 and osterix in bone tissue of the rats in osteoporosis group were significantly decreased (P<0.01),and the gene expression level of RANKL was increased(P<0.01); compared with osteoporosis group,the gene expression levels of PERK,ATF4 Runx2 and osterix in bone tissue of the rats in osteoporosis treatment group were significantly increased (P<0.01),and the gene expression level of RANKL was significantly decreased (P<0.01).Compared with control group,the protein expression levels of PERK,Runx2 and osterix in bone tissue of the rats in osteoporosis group were significantly decreased (P<0.05 or P<0.01),and the protein expression level of RANKL were increased(P<0.05 or P<0.01); compared with osteoporosis group,the protein expression levels of PERK,Runx2 and osterix in bone tissue of the rats in osteoporosis treatment group were significantly increased (P<0.05 or P<0.01),and the protein expression level of RANKL was significantly decreased (P<0.01).The HE staining results showed that compared with control group,the bone resorption pits in bone tissue of the rats in osteoporosis group became large with the increased bone absorption,which caused bone loss; compared with osteoporosis group,the resorption in bone tissue of the rats in osteoporosis treatment group was decreased,and the bone structure returned to normal. Conclusion: After the female rats are ovariectomized and injected with estrogen,the expression trends of PERK and osteoblast transcription factors Runx2 and osterix are consistent,in contrast with the osteoclast transcription factor RANKL expression,suggesting that the osteoblast function is reduced and it is related to the decreased expression of PERK in PMOP onset.

Objective:To investigate the role of MicroRNA-210(miR-210) in the growth process of ovarian cancer cells and its relationship with radiosensitivity, and to elucidate the effect of miR-210 on the development and treatment of ovarian cancer the possible molecular mechanism. Methods: The human ovarian cancer OVCAR3 and SKOV3 cells were cultured in vitro, miR-210 mimic and anti miR-210 inhibitors were transfected into the human ovarian cancer cell lines OVCAR3 and SKOV3, respectively, by cell transfection, and identified by Real-time PCR method; miR-210 overexpression and low expression of ovarian cancer cell models were obtained. The two kinds of cells were divided into control group, miR-210 overexpression model group and miR-210 low expression model group, and the cell proliferation activity was detected by MTT. The cells in control group and miR-210 overexpression model group were irradiated with different doses (30,50, and 100 Gy) of ionizing radiation, and the cell proliferation activity in each group was detected by MTT method. Western blotting method was used to detect the expression levels of apoptosis-related proteins in the ovarian cancer cells exposed to 50 Gy radiation dose. All the experimental cells were cultured with three double holes and repeated three times. Results: Compared with control group, the proliferation activity of the cells in miR-210 overexpression ovarian cancer cells group was enhanced, and the proliferation activity of the cells in miR-210 low expression cells group was reduced (P<0.05). After ionizing radiation, the proliferation activity of ovarian cancer cells in miRNA-210 overexpression group was enhanced compared with control group(P<0.05); the expression levels of apoptosis-related protein BAX in OVCAR3 and SKOV3 cells were significantly decreased, while the expression level of BCL-2 were significantly increased. Conclusion: miR-210 can promote the growth of ovarian cancer cells, and reduce the sensitivity of ovarian cancer cells to radiation therapy by inhibiting the apoptosis.

Objective:To discuss the influence of Tongfengning Capsule(TFN)in the rats with acute gouty arthritis,and to clarify the curative effect and mechanism of TFN in the acute gouty arthritis rats. Methods: The rat model of acute gouty arthritis was induced by injection of microcrystal sodium urate solution into articular cavity of the ankle joint.A total of 48 mice were divided into blank control group,model group,low(100 mg·kg^-1),middle(200 mg·kg^-1)and high(550 mg·kg^-1)doses of TFN groups,colchicine (0.63 mg·kg^-1) positive drug control group; 8 rats in each group.The degrees of joint swelling of the rats were monitored at different time after modeling.The number of white blood cells(WBC),lymphocytes and neutrophils in the whole blood of the rats in various groups were detected,and the nitric oxide(NO)levels in the serum and homogenate as well as serum uricacid(UA)were detected.The levels of IL-1β and TNF-α in the soft tissue and synovial fluid around the ankle joint of the rats in various groups were measured. Results: Compared with model group,the swelling degrees of ankle joint of the rats in positive drug control group and high dose of TFN group were significantly decreased in 0-8 h (P<0.05 or P<0.01),especially in 4 h(P<0.01);but there were no significant differences in low and medium doses of TFN groups (P>0.05).Compared with model group,the levels of WBC,lymphocytes and neutrophils in the blood of the rats in positive drug control group and TFN groups were decreased,especially in high dose of TFN group (P<0.05).The UA levels in serum of the rats in positive drug control group and high dose of TFN group were significantly lower than that in model group (P<0.05 or P<0.01),and the levels of serum NO was increased (P<0.05);but there were no significant differences in low and medium doses of TFN groups(P<0.05).The levels of NO,IL-1β and TNF-α in soft tissue and synovial fluid of the rats in positive drug control group and high dose of TFN group were significantly lower than those in model group(P<0.05 or P< 0.01);but there were no significant differences in low and medium doses of TFN groups(P<0.05). Conclusion: TFN has a remarkable effect in the treatment of gouty arthritis,and this effect may be associated with inhibiting the inflammatory reactions.

Objective:To observe the morphological features of liver injury induced by hydrodynamic injection,and to explore the procedure and mechanism of liver repair. Methods: Twenty-five female Balb/c mice were intravenously injected with large volume of saline solution (1.8-2.0 mL) in 3 s. The mice were divided into 30 min,8 h,1 d,3 d,and 7 d groups according to the post-injection time. Each group contained 5 mice. Six non-injected mice were used as control group. The blood samples from angular vein of the mice were collected to detect the levels of alanine transaminase (ALT).The morphological features of liver tissue of the mice in various groups were observed with HE staining.The proliferating cell nuclear antigen (PCNA) protein expressions in liver tissue of the mice in various groups were detected by immunohistochemical saining.The expression levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),epidermal growth factor (EGF),hepatocyte growth factor (HGF),transforming growth factor-α (TGF-α),Cyclin D1,vascular endothelial growth factor (VEGF),Bax and Bcl-2 mRNA were determined by Real-time PCR. Results: Compared with control group,the ratio of liver weight/original body weight in 8 h group was significantly decreased (P<0.05),but there was no significant difference in 7 d group (P>0.05).Compared with control group,the serum ALT level in 1 d group was significantly elevated (P<0.01),but there was no significant difference in 7 d group (P>0.05).Compared with control group,the liver tissue of the mice in 30 min group showed hepatocyte swollen and small hemorrhagic area,and the number of hepatocytes per high power field was significantly declined (q=4.760,P<0.05) under microscope. Compared with 30 min group,the number of swollen cells was decreased in 8 h group,the hemorrhagic and necrotic area enlarged,both of the hepatocyte number and binuclear hepatocyte number per high power field were significantly increased (q=7.310,P<0.01;q=7.200,P<0.01).Compared with 8 h group,the hemorrhagic and necrotic areas in 1 d group were reduced,both of the hepatocyte number and binuclear hepatocyte number per high power field were significantly decreased (q=4.966,P<0.05;q=6.596,P<0.01); there were no significant differences compared with control group (P>0.05).The hemorrhagic and necrotic areas almost disappeared in 3 d group.The histological appearance of liver tissue of the mice in 7 d group was same as control group.Compared with control group,the PCNA indexes in hepatocytes in 8 h and 1 d groups were significantly increased (t=4.458,P<0.01; t=15.557,P<0.01); there was no significant difference in 7 d group (P>0.05).Compared with control group,the PCNA index in cholangiocytes in 30 min group was increased significantly (t=3.985,P<0.01); there was no significant difference in 7 d group (P>0.05).The mRNA expression levels of TNF-α,IL-6,EGF,and VEGF in liver tissue of the mice in 30 min group were significantly higher than those in 8 h group (q=4.952,P<0.05; q=14.750,P<0.01; q=14.750,P<0.01; q=13.551,P<0.01).The HGF mRNA expression level in liver tissue of the mice in 3 d group was significantly higher than those in 30 min group and 8 h group (q=5.031,P<0.05;q=4.631,P<0.05). The TGF-α mRNA expression levels in liver tissue of the mice in 8 h group and 1 d group were higher than that in 30 min group (q=4.592,P<0.05; q=8.137,P<0.01).The Cyclin D1 mRNA expression levels in liver tissue of the mice in 8 h group and 1 d group were higher than that in 7 d group (q=4.736,q=5.213,P<0.05). The Bax/Bcl-2 ratio of the mice in 1 d group was higher than that in 30 min group (q=5.731,P<0.01). Conclusion: Acute liver injury induced by hydrodynamic injection mainly shows the hepatocyte swelling and hemorrhagic necrosis.The injury could be repaired naturally in a week.The various cytokines and growth factors related with liver regeneration are involved in the repair procedure.

Objective:To simulate the pollution environment of indoor decoration volatile pollutants,and to explore the influence of three main indoor decoration volatile pollutants formaldehyde,xylene and ammonia in the male mouse reproductive function. Methods: A total of 20 male ICR mice were randomly divided into experimental group (exposed to 60 mg·m^-3 formaldehyde,50 mg·m^-3 xylene and 40 mg·m^-3 ammonia mixed in the static type canister) and control group (put into the air-filled static type canister) (n=10).Each mouse was weighed at set times every day,exposed for 35 d continuously.The behavior changes of the mice were also observed everyday.The epididymal cauda sperms were collected to detect the sperm density and vitality.The sperm malformation rate was detected by Diff-Quick dyeing method.The apoptotic rate of epididymal cauda sperms was detected by flow cytometry(FCM).Then Western blotting method was used to determine the expression levels of Caspase-9 and Cleaved Caspase-3 proteins in testis tissue of the mice. Results: Four weeks after the exposure to pollutants,the body weight of the mice in experimental group was significantly lower than that in control group (P<0.01).The concentration and vitality of sperm malformation rate of the mice in experimental group were significantly lower than those in control group (P<0.01) 35 d after exposure.Compared with control group, the apoptotic rate of sperm of the mice in experimental group was significantly increased (P<0.01).However,the expression levels of Caspase-9 and Cleaved Caspase-3 in the testis tissue of the mice in experimental group and control group had no significant differences (P>0.05). Conclusion: The main pollutants in indoor decoration can significantly influence the quality of spermatozoa in the epididymis.

Objective:To detect the repair effect of brain protein hydrolyzate (BPH) on the oxidative damage induced by H₂O₂ in the PC12 cells,and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method. Methods: The PC12 cells in the logarithmic phase were divided into normal control group, model group (300 μmol·L^-1 H₂O₂),BPH group,artificial gastric juice-treated BPH (GBPH) group,artificial intestinal juice-treated BPH (IBPH) group,artificial gastric juice-treated BPH tablets (GBPH-T) group and artificial gastric juice-treated BPH floating tablets (GBPH-FT) group (The latter five groups were added with 20,40,60,80 and 100 mg·L^-1 BPH treated with different conditions);at the same time blank control group was set up. The PC12 cells in normal control group didn't receive any treatment,the blank control group was added with medium only,and the PC12 cells in model group were treated with H₂O₂ (300 μmol·L^-1) for 3 h.The cell viability was detected by MTT assay.Using Design-expert 8.0.6 Trial software,the dosages of HPMC-K4M,octadecanol and acrylic resin Ⅱ were used as the investigation factors,and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription. Results: Compared with normal control group, the activity of PCl₂ cells in 60 mg·L^-1 BPH group was increased (P<0.05). Compared with model group,the vitalities of PC12 cells in BPH group,IBPH group,GBPH group and GBPH-FT group were increased significantly (P<0.05 or P<0.01). Compared with BPH group,the cell viability in IBPH group was decreased significantly (P<0.05),and there was no significant difference in GBPH group (P>0.05). Compared with GBPH-T group,the cell viability in GBPH-FT group was significantly increased (P<0.05).The optimal prescription was BPH 20 mg,lactose 10 mg,magnesium stearate 0.4 mg,microcrystalline cellulose 20 mg,HPMC-K4M 27 mg,octadecanol 63 mg,acrylic resinⅡ13 mg; all floating tablets drift in 5 s and sustained floating > 8 h;the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant(P>0.05). Conclusion: BPH has a significant repair effect on the H₂O₂-induced oxidative damage in the PC12 cells. Star design-effect surface method has good predictability and reproducibility;it is reasonable and feasible,and can be used to optimize the prescription of BPH stomach floating tablets.

Objective:To explore the inhibitory effects of gomisin A (GA) in combination with carboplatin (CBP)on the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells and their mechanisms,and to illustrate the synergistic antitumor effect of GA. Methods: The ovarian cancer Skov3 cells were treated with different concentrations of GA combined with different concentrations of CBP.MTT assay was used to detect the inhibitory rates of proliferation.According to the results,the appropriate concentrations of GA and CBP were conformed.The Skov3 cells were treated with PBS,GA(0.04 μmol·L^-1),CBP (16 mg·L^-1) and GA (0.04 μmol·L^-1) in combination with CBP (16 mg·L^-1),respectively, and used as control group,GA group,CBP group and GA+CBP group. After 48 h of treatment,the cell morphology was observed by optical microscope,the cell reproductive ability was determined by clonogenic cell survival assay,and the cell migration ability was measured by cell wound healing test;Transwell experiment was used to detect the cell invasion ability.The expression levels of MMP-2 and MMP-9 mRNA were determined by RT-PCR and the protein expression levels of MMP-2,MMP-9,AKT1 and pAKT1 were determined by Western blotting method. Results: Compared with GA and CBP groups,the inhibitory rate of proliferation of Skov3 cells in GA + CBP group was significantly increased(P<0.01).The concentrations of GA and CBP for the best dose ratio were 0.04 μmol·L^-1 and 16 mg·L^-1,respectively. The results of clone formation assay showed that the colony formation rates in CBP and GA+CBP groups were decreased compared with control group(P<0.05 or P<0.01);the colony formation rate of cells in GA + CBP group was significantly decreased compared with GA and CBP groups(P<0.01); the wound scratch assay results suggested that the number of metastic Skov3 cells in GA and CBP groups were decreased compared with control group;and the number of metastic Skov3 cells in GA + CBP group was decreased compared with GA and CBP groups.The results of Transwell expriment showed that the capacities of cells passing the ECM in GA and CBP groups were decreased compared with control group;the capacity of cells passing the ECM was decreased compared with GA and CBP groups.The RT-PCR and Western blotting results showed that the expression levels of MMP-2 and MMP-9 mRNA in GA+CBP group were down-regulated compared with GA and CBP groups(P<0.01),and the expression levels of MMP-2,MMP-9,AKT1 and pAKT1 protein were down-regulated(P<0.01). Conclusion: GA can enhance the ability of CBP to inhibit the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells,and thus play a role in decreasing the toxicity and increasing the efficacy of chemotherapy.

Objective:To investigate the MUC1 specific immune response enhanced by thmosinα1(Tα1) using MUC1-MBP as the specific antigen,and to discuss the feasibility of MUC1-MBP as an adjuvant. Methods: The C57BL/6 mice were randomly divided into normal saline group,MUC1-MBP+BCG group and MUC1-MBP+Tα1 group.The T cellular immune activities,MUC1 special antibody and subclass,anti-tumor effect of Tα1 combined with MUC1-MBP were detected.4-7d after the 3rd immunization,the spleen indexes of the mice in various groups were measured; the lymphocyte proliferation response was used to detect the stimulate index(SI) of the mice in various groups; the levels of specific cytokines IFN-γ,IL-2,IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA; the level of serum MUC1-specific antibody was detected by ELISA.The C57BL/6 mice were inoculated with B16-MUC1 7 d after the last immunization and the survival of the mice was observed. Results: Compared with normal saline group,the spleen index and SI of the mice in MUC1-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased (P< 0.05 or P<0.01); the specific SI of MUC1 were significantly increased (P< 0.05).Compared with normal saline group,the levels of IFN-γ and IL-2 in the supernatant of spleen cells of the mice in MUC1-MBP+BCG group were obviously increased (P<0.05);the levels of IL-4 and IL-10 were slightly increased,but there were no significant differences (P>0.05); compared with normal saline group,the level of IL-4 in MUC1-MBP+Tα1 group was obviously increased (P< 0.01);the levels of IFN-γ,IL-2 and IL-10 were slightly increased,but there were no significant differences (P>0.05). The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1. The antibody subtype detection results showed that compared with normal saline group,the level of IgG1 in MUC1-MBP+BCG group was significantly increased(P< 0.05 or P< 0.01),and the level of IgG2a had no obvious change.The tumor prevention experiment results showed that compared with normal saline group,the survival rates of the mice in MUC1-MBP+BCG group and MUC1-MBP+Tα1 group had no significant differences. Conclusion: MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice.It indicates that Tα1 can be used as an adjuvanted preventive vaccines,but not suitable for therapeutic vaccines.

Objective:To investigate the effect of Schisandra chinensis polysaccharide (SCP) on the growth of brain tumor stem cells (BTSCs), and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods: The primary human glioma cells were cultured, then the BTSCs were isolated by CD133 immunomagnetic sorting. The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay. The proliferation rate of BTSCs was examined by MTT assay. Annexin Ⅴ-PI analysis was used to analyze the apoptotic rate of BTSCs. The expression levels of Bax, Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay. Results: The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs. Compared with control group, the proliferation rates of BTSCs in 200, 400 and 800 mg·L^-1 SCP groups were decreased, especially in 400 and 800 mg·L^-1 SCP groups (P<0.05). The results of Annexin Ⅴ-PI analysis showed that the apoptotic rate of BTSCs in 800 mg·L^-1 SCP group was increased compared with control group (P<0.05). The ELISA results showed that the expression levels of Bax in 200, 400 and 800 mg·L^-1 SCP groups were significantly increased (P<0.05), and the values of Bax/Bcl-2 were significantly increased (P<0.05); compared with control group, the Bcl-2 expression level in the BTSCs in 800 mg·L^-1 SCP group was decreased (P<0.05). The expression level of Caspase-3 protein in 800 mg·L^-1 SCP group was also significantly increased compared with control group (P<0.01). Conclusion: SCP could inhibit the growth of BTSCs,and the induction of apoptosis may be one of mechanisms.

Objective:To set up the rat skeletal muscle L6 cell models of oxygen-glucose deprivation (OGD) in vitro,and to investigate the protective effect of EGF in deep tissue injury(DTI) of pressure sores. Methods: The rat skeletal muscle cells in the logarithmic phase were divided into normal control group,OGD group,5 μg·L^-1 EGF+OGD group,10 μg·L^-1 EGF+ OGD group and 20 μg·L^-1 EGF+ OGD group.The survival rates of skeletal muscle cells in various groups were measured by MTT assay; the cell apoptotic rates in various groups were detected by flow cytometry; the reactive oxygen species (ROS) levels were detected by DCFH-DA; Rhodamine 123 was used to detect the mitochondrial membrane potential; the expressions of Bax and Bcl-2 proteins were determined by Western blotting method. Results: Compared with normal control group,the survival rates of skeletal muscle cells in OGD group after 24 h OGD was significantly decreased (P<0.05); the apoptotic rate was markedly increased (P<0.01); the ROS level was increased (P<0.01);the mitochondrial membrane potential was decreased (P<0.01); the ratio of Bcl-2/Bax was significantly decreased (P<0.01).Compared with OGD group, the survival rates of skeletal muscle cells in different concentrations of EGF groups were increased and the apoptotic rates were decreased,especially in 10 and 20 μg·L^-1 EGF groups (P<0.05 or P<0.01); the ROS levels in skeletal muscle cells in different concentrations of EGF groups were decreased and the mitochondrial membrane potential were increased,especially in 10 and 20 μg·L^-1 EGF groups (P<0.05 or P<0.01); the Bcl-2/Bax ratios were significantly decreased in a concentration-dependent manner,especially in 10 and 20 μg·L^-1 EGF groups(P<0.05 or P<0.01). Conclusion: EGF can improve the skeletal muscle cell injury induced by OGD in a concentration-dependent manner via decreasing the ROS levels and protecting the cell mitochondrial function.

Objective:To investigate the effects of silencing HOXA13 gene on the malignant phenotypes of hepatocellular carcinoma cell lines HepG₂ and QGY-7703,and to provide a new molecular target for the diagnosis and treatment of hepatocellular carcinoma. Methods: The interference recombinant plasmid of plent-U6-GFP/si-HOXA13 was constructed,then it was stably transfected into the HepG₂ and QGY-7703 cells.The interfering effects of HOXA13 gene were determined by RT-PCR and Western blotting methods.The HepG₂ and QGY-7703 cells transfected with HOXA13 knockdown were used as experimental group,and the HepG₂ and QGY-7703 cells transfected with empty plasmid were used as control group.Then the growth speed was examined by MTT assay,the cell doubling time was examined by double time assay,the colony formation ability was examined by colony formation assay,and the cell cycle was examined by flow cytometry. Results: The MTT assay results showed that compared with control group,the growth speeds of HepG₂ and GY-7703 cells in experimental group were significantly decreased and the cell cycles were changed; the number of HepG₂ and QGY-7703 cells in G₁ phase was increased and the number of cells in S phase was decreased.The doubling time of HepG₂ and QGY-7703 cells in control group and experimental group were (4.59±0.27),(4.93±0.17),(6.02±0.86), and (6.43±0.66)h,and the differences between control group and experimental group were significant(P<0.05).The colony number of HepG₂ and QGY-7703 cells in control group and experimental group were 264.00±12.62,269.00±4.55,165.00±10.61, and 215.00±4.43,and the differences between control group and experimental group were significant(P<0.01). Conclusion: HOXA13 can increase the proliferation,enhance the clone formation,decrease the number of cells at G₁ phase and increase the number of cells at S phase in the hepatocellular carcinoma HepG₂ and QGY-7703 cells; and it may used as a new molecular target for diagnosis and treatment of hepatocellular carcinoma.

Objective:To study the effects of resveratrol on the expressions of Caspase-3 and Bcl-2 in the retina tissue of the rats with retinal ischemia-reperfusion injury (RIRI),and to investigate the therapeutic effect of resveratrol on the RIRI and its mechanism. Methods: A total of 90 SD rats were randomly divided into sham operation group,model group and treatment group,30 rats in each group.The RIRI models were established by pressing the anterior chamber of the rats.The rats in model and treatment groups received ischemia-reperfusion for 1,6,12,24,and 48h;the rats in treatment group were treated with micro-syringe intravitreal injection of 0.5 nmol·L^-1 of resveratrol 5 μL.The retinal tissue structure was observed by inverted microscope.The expression levels of Caspase-3 and Bcl-2 in retina tissue were detected by immunohistochemistry and Western blotting methods. Results: The retinal tissue edema of the rats in model group was found with vacuolar degeneration of the ganglion cells,and the arrangement of the cell layer was loose;the number of retinal ganglion cells was decreased,the boundary was blurred,and the nerve fiber layer thinned obviously.The degree of retinal tissue structure,the degree of injury and the degeneration of ganglion cells in treatment group were lighter than those in model group.The results of immunohistochemistry showed that the expressions of Caspase-3 and Bcl-2 in the retina tissue of the rats in treatment group were significantly increased compared with model group.The results of Western blotting method showed that the expression levels of Bcl-2 protein in the retina tissue of the rats in treatment group at different time points were increased compared with model group;and there were significant differences at 24 and 48 h (P<0.05);the expression levels of Caspase-3 protein in treatment group at different time points were lower than those in model group (P<0.05). Conclusion: Resveratrol can improve the retinal tissue structure of the RIRI rats,and its mechanism may be related to decreasing the expression level of Caspase-3 and increasing the expression level of Bcl-2 in the retinal tissue.

Objective:To investigate the therapeutic effects of small peptide, alkaloid, total flavonoids and polysaccharides extracted from sunflower plate powder on the hyperuricemia in the mice, and to clarify that the influence of sunflower powder in the uric acid (UA) level and joint swelling and its protective effect on the liver and kidney tissues. Methods: A total of 96 male Kunming rats were randomly divided into 8 groups, each group of 12 rats.One group was used as blank control group,the others were used to set up hyperuricemia models. The successful mouse models of hyperuricemia were randomly divided into model group and positive control group (allopurinol group), small molecule peptide group, alkaloi group, total flavone group, polysaccharide group and sunflower disk group. The serum UA levels were measured 7 d after administration and the liver and kidney tissues of the mice in various groups were taken. The morphological changes of liver and kidney tissues were observed by HE staining.Acute gouty arthritis model was established by injecting 3 mg sodium urate into the knee joint of the male SD rats. The experiment was divided into blank group, model group, positive control colchicine group, sunflower disc extracts (SDE) group with different concentrations (0%, 20%, 40%, 60%, 80% and 100%),4 rats in each group. The joint circumference was measured at 0, 12, 24 and 48 h, respectively. The serum levels of interleukin-10(IL-10) and macrophage inflammatory protein-1α(MIP-1α) were detected by ELISA kit. Results: Compared with model group, the serum UA levels of the mice in allopurinol group, small molecule peptide group, alkaloid group, total flavonoids group and sunflower group were significantly decreased after administration (P<0.05); and the serum UA level in allopurinol group was decreased the most, followed by small molecule peptide group.The HE staining results showed that the hepatocytes in small molecule peptide group were clear,the hepatocytes had full cytoplasm, and no abnormality was observed in the kidney cells compared with blank control group. After 48 h injection of sodium urate, the degree of joint swelling was reduced by up to 27.1% in SDE group compared with model group. Compared with model group, the serum IL-10 level in 20% SDE group was significantly increased (P<0.01), and the serum MIP-1α level was also significantly increased (P<0.01). Conclusion: Sunflower powder can reduce the UA level and has protective effect on the liver and kidney tissues and it can improve the body's anti-inflammatory activity and reduce the body's inflammatory response to achieve anti-gout arthritis activity.

Objective:To study the dynamic characteristics of reconstruction of B lymphocytes in peripheral blood at different time points in the patients with immune tolerance induced by combined transplantation of kidney and bone marrow,and to provide the basis for the further illumination of the role of B lymphocytes in immune tolerance of organ transplantation. Methods: Five patients (Pt.A,Pt.B,Pt.C,Pt.D,and Pt.E)with end stage of kidney disease received kidney and bone marrow transplantation after induction therapy.Immunosuppression was discontinued gradually.The number of B lymphocytes and the distribution of subsets of B lymphocytes in peripheral blood were detected by flow cytometry in the patients before and after transplantation,and the dynamic changes of B lymphocytes at different time points were compared.The distribution of immunoglobulin heavy chain variable region(IGHV) was analyzed using next generation sequencing technology. Results: Four patients were enrolled in the study expect one patient with graft failure(Pt.C).The B lymphocyte counts of Pt.A,Pt.B and Pt.D recovered approximately 1 year after transplantation and Pt.E experienced delayed reconstitution.The B lymphocyte recovery was accompanied by a high frequency of CD20+CD24^high CD38^high transitional B lymphocytes and a diversified clonal repertoire.All patients showed the prevalence of CD20+CD27+ memory B cells around 6 months after transplantation.Through the calculation of Shannon diversity index (SDI) of IGHV at each time point after transplantation,the SDI of IGHV for all transplant recipients at 182 d after transplantation was significantly lower than before transplantation (P=0.004),and this index was gradually recovered at approximately 1 year after transplantation.The calculation of somatic mutation of B lymphocyte IGHV gene sequence at different time points showed that the somatic mutation rate was elevated at 182 d for all patients except Pt.B (P=0.032). Conclusion: The presence of mutated memory B lymphocytes in peripheral blood of the patients with kidney and bone marrow transplantation is found,and the B lymphocytes play the potential contribution to tolerance induction.

Objective:To explore the differences of the relative expression levels of plasma long chain non encoding RNA (lncRNA) MALAT1, LincRNA-p21 and GAS5 in the patients with multiple myeloma (MM) and chronic lymphocytic leukemia (CLL), and to clarify their significances in the differential diagnosis of MM and CLL. Methods: A total of 60 cases of MM patients (MM group), 60 cases of CLL patients (CLL roup) and 60 healthy persons after physical examinations (control group) were selected as the subjects. The plasma levels of lncRNA MALAT, LincRNA-p21 and GAS5 of the subjects in three groups were detected and compared. Results: The relative expression levels of lncRNA MALAT1 and GAS5 in plasma of the patients in MM group were significantly higher than those in the other two groups(P<0.05);the relative expression level of LincRNA-p21 in plasma of the patients in CLL group was significantly lower than those in the other two groups(P<0.05).The area under receiver operating characteristic curve(ROC)(AUC) of plasma LincRNA-p21 in the diagnosis of CLL was 0.850, its 95% confidence interval (CI) was 0.780-0.921. The AUC of plasma lncRNA MALAT1 and GAS5 in the diagnosis of MM were 0.898 and 0.815; their 95% CI were 0.836-0.959 and 0.740-0.890, respectively. The AUC of plasma lncRNA MALAT1, LincRNA-p21 and GAS5 in the differential diagnosis of CLL and MM were 0.878, 0.778 and 0.805, and their 95%CI were 0.814-0.942, 0.691-0.865 and 0.727-0.882, respectively. Conclusion: The incidence of MM is related with the high expressions of lncRNA MALAT1 and GAS5 in plasma and the incidence of CLL is related with the low expression of LincRNA-p21 in plasma. The relative expression levels of the above lncRNA can be used as the auxiliary indexes in the diagnosis of MM and CLL.

Objective:To investigate the expressions of hypoxia-inducible factor-1α (HIF-1α) and heat shock protein 70 (HSP70) in the placenta tissue of the patients with pregnancy-induced hypertension syndrome(PIH),and to elucidate the clinical significances of their expressions in the placenta tissue of the patients with PIH. Methods: Twenty-two cases of placenta tissue of the PIH patients (PIH group) and eighteen cases of placenta tissue of the normal pregnant women (control group) were selected. The expressions of HIF-1α and HSP70 were detected by immunohistochemical method.The differences of potive expressions rates of HIF-1α and HSP70 in placenta tissue of the PIH patients and the normal pregnant women and their relationships in placenta tissue of the PIH patients were analyzed. Results: The positive expression rate of HIF-1α in placenta tissue of the patients in PIH group(81.8%) was higher than that of the normal pregnant women in control group(38.9%) (χ²=7.785,P=0.005),and the positive expression rate of HSP70 in placenta tissue of the patients in PIH group (90.9%) was higher than that of the normal pregnant women in control group(55.6%) (χ²=6.599,P=0.010). Eighteen cases of placenta tissue with HIF-1α positive expression in the PIH patients had HSP70 positive expression (100.0%);among four cases of placenta tissue with HIF-1α negative expression, two cases had HSP70 negative expression (50.0%); their expressions in placenta tissue of the PIH patients had positive correlation(r=0.671,P=0.001). Conclusion: The expressions of HIF-1α and HSP70 in placenta tissue of the PIH patients are higher,and they have positive correlation.

Objective:To explore the related influencing factors of carotid atherosclerosis (AS) plaque formation and stability in the patients with type 2 diabetes mellitus (T2DM), and to provide scientific evidence for early prevention and treatment. Methods: A total of 249 cases of simple T2DM patients were selected. According to the results of carotid artery color Doppler ultrasound, they were divided into AS group, stable plaque group and unstable plaque group. The differences in physicochemical indexes and life style of the patients were compared between three groups; univariate analysis and Logistic regression analysis were used to screen the related influencing factors of carotid AS plaque formation and stability of the T2DM patients. Results: The univariate analysis showed that age, fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), non-high density lipoprotein cholesterol (NHDL-C), monocyte/high density lipoprotein ratio (MHR), smoking ratio(Smoking%), T2DM disease course, high density lipoprotein protein cholesterol (HDL-C) had significant differences between AS group and plague group (P<0.05). There were significant differences in the values of postprandial blood glucose (PBG), systolic blood pressure (SBP), low density lipoprotein cholesterol (LDL-C), HbA1c, MHR, Smoking% and HDL-C of the patients in stable plaque group and unstable plaque group (P<0.05). The Logistic regression analysis showed that high age, HbA1c, NHDL-C, MHR and smoking were the risk factors of the plaque formation and high HDL-C was a protective factor of plaque formation (Age:OR=1.62, P=0.011; HbA1c:OR=1.25,P=0.027; HDL-C:OR=0.65, P=0.009; MHR:OR=3.50, P=0.000; Smoking:OR=2.28, P=0.009; NHDL-C:OR=1.39, P=0.028).High SBP, LDL-C, MHR and smoking were the risk factors of unstable plaque formation, and high HDL-C was a protective factor of unstable plaque formation(SBP:OR=1.57,P=0.003;LDL-C:OR=1.99,P=0.000;MHR:OR=3.88,P=0.000;Smoking:OR=2.01,P=0.001;HDL-C:OR=0.53,P=0.001). Conclusion: For the patients with simple T2DM and carotid AS plaque, blood lipid, blood pressure and smoking cessation should be emphasized and HDL-C level should be increased, which can effectively prevent the formation of AS plaque and stabilize the plaque.

Objective:To investigate the differences in the curative effects of neoadjuvant chemotherapy (NAC) for different subtypes of Luminal B breast cancer and its prognosis, and to discuss the clinical treatment characteristics of different subgroups. Methods: A total of 246 cases of Luminal B like breast cancer patients who completed the projected NAC courses and surgical treatment were selected. All the biopsy specimens before treatment were positive for estrogen receptot(ER). According to the expressions of progesterone receptor(PR), human epidermal growth factor-2(Her-2) and cell proliferation nuclear antigen Ki-67, 246 cases of Luminal B breast cancer patients were divided into 3 subgroups. A subgroup (PR low expression group), Her-2 negative and PR< 20% or negative, Ki-67 any levels; B subgroup (PR high expression group), Her-2 negative, PR ≥ 20% and Ki-67 ≥ 14%; C subgroup (Her-2 positive group), Her-2 positive, Ki-67 and PR any levels. The clinical pathological materials and follow-up recurrence events of the patients were collected. Results: Among the three subtypes of Luminal B breast cancer, there were no significant differences in the age, the size of primary tumor and the stage of TNM (P>0.05). There were significant differences in the lymph node metastasis rate among three subgroups (P=0.018), and the lymph node metastasis rate was the highest in A subgroup among three subgroups. There were no significant differences in the clinical response and pathological response among three subgroups (P=0.123, P=0.06). 8.5% (21/246) patients achieved the pathological complete response (pCR); the rates of pCR in three subgroups had statistically significant difference (P=0.009); the rate of pCR in C subgroup was the highest, and the rate of pCR in B subgroup was the lowest. The Log-Rank test of the survival curves of three subgroups had not statistically significant difference (P=0.216), but the 3-year disease-free survival(DFS) and 5-year DFS of the patients in B subgroup were slightly higher than those in other two groups. The DFS of the patients in C subgroup was longer than that of the patients with Her-2 overexpression breast cancer at the same period, and the difference was statistically significant (P=0.047). Conclusion: Her-2 positive Luminal B breast cancer is more likely to achieve pCR in NAC and the prognosis is better than Her-2 overexpressing breast cancer. The patients with high expression of PR in Luminal B breast cancer patients have a tend of overall survival advantage compared with the patients with PR low expression and Her-2 positive expression.

Objective:To explore the differences in the positive rates of specific-IgE(sIgE) of the common allergens between the cow's milk protein allergy (CMPA) and healthy infants and the distribution characteristics of positive sIgE of common allergens in the CMPA infants,and to provide basis for comprehensive intervention of the CMPA infants. Methods: A total of 156 cases of CMPA and 318 cases of healthy infants were selected as the subjects. The serum sIgE and total IgE levels of common allergens of the infants in two groups were detected by enzyme-linked immunosorbent assay. The differences in the positive rates of serum sIgE of common allergens and total IgE of the infants in two groups were compared. Results: There were no statistical differences in the positive rates of serum sIgE and total IgE for common allergens of the infants between CMPA group and healthy group(P>0.05).The major food allergens with high positive rates of sIgE in CMPA group were cow's milk(44.2%),egg white(10.3%) and cashew nut(5.1%), and the inhale allergens with high positive rates of sIgE were cat hair(21.2%), dog hair(9.6%) and house dust mite(4.5%).While for the healthy infants, the major food allergens with high positive rates of sIgE were cow's milk(45%),egg white (14.2%)and cashew nut(6.0%),and the inhale allergens with high positive rates of sIgE were cat hair(25.8%),dog hair(14.5%)and fungus combinations(4.5%). The analysis in different age groups (<1 year old and 1-2 years old) showed that there were no statistical differences in the positive rates of serum sIgE and total IgE for common allergens of the infants between CMPA group and healthy group(P>0.05). Conclusion: There are no significant differences in the serum sIgE and total IgE positive rates between the CMPA and healthy infants. The detection of serum sIgE and total IgE of common allergens is of little clinical significance for the CMPA infants.

Objective:To investigate the efficacy and side effects of combination of methylprednisolonepulse(MDP) and mycophenolate mofetil(MMF) in the treatment of systemic lupus erythematosus(SLE) in the children. Methods: A total of 16 cases of children with SLE,lupus nephritis(LN) and type Ⅳ diffuse glomerular mesangial proliferative glomerulonephritis diagnosed by pathology were selected.Among them 7 cases were given MDP combined with MMF,and received intermittently oral small dose of corticosteroids(GC), and they were used as pulse therapy group; 9 cases were given oral GC and transitional reduction,and they were used as traditional therapy group. SLEDAI was used for the evaluation of the curative effect,and body mass index(BMI),blood pressure(BP),intraocular pressure(IOP),triglycerides(TG),fasting blood glucose(FBG) and serum calcium(Ca) were analyzed during the treatment of 1 year and then the efficacies and side effects of the children in two groups were compared. Results: The SLEDAI scores,levels of complements C3 and C4,24 h urinary protein outcome of the children in pulse therapy group were better than those in traditional therapy group; the differences in SLEDAI scores were statistically significant after treating for 3 and 6 months between two groups(P<0.05); ESR and 24 h urinary protein outcome had significant differences after 6 months of treatment between two groups(P<0.05); the complement C3 difference was statistically significant between two groups(P<0.05) after 12 months of treatment.Compared with traditional therapy group,the BMI,IOP,TG,FBG of the children in pulse therapy group after treatment were decreased(P<0.05);the BMI,IOP and TG had significant differences after treating for 12 months between two groups(P<0.05);the differences in FBG were statistically significant after treating for 6 and 12 months between two groups(P<0.05).The Ca of the patients in pulse therapy group was higher than that in traditional therapy group,but there was no statistically significant difference(P>0.05). The SPB and DBP of the patients in pulse therapy group were higher than those in traditional therapy group,but the differences were not statistically significant(P>0.05).At the same time,gastrointestinal ulcers,bleeding,perforation,pancreatitis,cardiovascular events (such as cardiac arrhythmias) didn't occur in two groups. Conclusion: Compared with traditional therapy,the combined treatment of MDP and MMF can control the symptoms of SLE early and rapidly,and reduce the viscera damage.To choose 1 year after treatment as observation point,its disease activity is lower than the traditional therapy,and the curative effect is better than oral GC transitional reduction with immunosuppressant therapy.The GC-related side effects are lower than traditional therapy.

Objective:To discuss the curative effect of slow stretching training combined with extracorporeal shock wave in the treatment of spasticity of biceps brachii in the stroke patients, and to provide the clinical evdiences for the application of this therapy. Methods: Fifty-six patients with post-stroke biceps bachii spasticity were randomly divided into observation group (n =28) and control group (n=28) according to random number table method. The patients in two groups received routine treatment (40 min/time, 2 times·d^-1, 6 d per week) and slow stretching training (15 min/time, 2 times·d^-1, 6 d per week). On the basis of the routine treatment, the patients in observation group were treated with extracorporeal shock wave, while the patients in control group were treated with pseudo-extracorporeal wave therapy. Modified Ashworth scale (MAS), simplified upper limb Fugl-Meyer score (FMA) and modified Barthel index (MBI) were used to evaluate the curative effects before treatment, 2 weeks and 4 weeks after treatment. Results: After 2 weeks of treatment, the FMA score of the patients in observation group was siginificantly increased compared with before treatment (P<0.05), and the MBI and MAS scores were decreased (P<0.05); the MAS score of the patients in observation group was significantly lower than that in control group (P=0.036),while there were no significant differences in the FMA and MBI scores between two groups (P>0.05). Compared with 2 weeks after treatment, the MAS score of the patients in observation group 4 weeks after treatment was significantly decreased (P<0.05), and the FMA and MBI scores were increased (P<0.05); the FMA score and the MBI score in observation group were significantly higher than those in control group (P<0.05 or P<0.01), and the MAS score was significantly lower than that in control group (P<0.01). Conclusion: Slow stretching training combined with extracorporeal shock wave could effectively improve the post-stroke biceps brachii spasticity, and its therapeutic effect is better than the simple application of slow stretching training.

Objective:To observe the curative effect of 25-gauge mini-invasive system in cataract surgery after vitrectomy,and to provide a safe operation method for the patients with cataract induced by vitrectomy. Methods: A total of 29 patients (30 eyes) with cataract after vitrectomy were selected. Phacoemulsification and implantation of artificial lens with 25-gauge mini-invasive irrigation system were performed in the patients.The best corrected visual acuity(BCVA),intraocular pressure(IOP),corneal endothelial cell loss rate,intraoperative and postoperative complications of the patients were observed. Results: All the patients received the operation successfully. One month after operation,BCVA ≥ 0.5 were found in 5 eyes,0.3-0. 5 in 8 eyes,0.1-0.3 in 9 eyes,Fc-0.1 in 8 eyes; the BCVA 1 month after operation had significant differences compared with before operation(P<0.05).The mean IOP before operation and 1 d,7 d,1 month,3 months after operation were (17.13±2.84),(17.20±4.16),(17.43±3.41),(17.36±2.51),and(16.73±2.33)mmHg;the IOP of the patients at 1 d after operation had significant difference compared with before operation(P<0.05).No statistically significant differences of IOP were found between other groups(P>0.05).The number of corneal endothelial cells before operation was(2 467±280)mm^-2 and the number after operation was (2 033±209)mm^-2;the loss rate of corneal endothelial cells was 17.59%.There were no sever complications such as retinal redetachment,detachment of choroid,capsular rupture or suprachoroidal hemorrahage. Conclusion: Phacoemulsification for cataract after vitrectomy with 25G mini-invasive system way can maintain the IOP,reduce the complications and enhance the security in cataract surgery.

Objective:To compare the clinical effects of different minimally invasive surgeries in the treatment of varicose veins of lower extremities,and to explore their application values. Methods: A total of 201 patients with varicose veins of lower extremities were selected and treated with different operation methods.52 cases were treated by endovenous laser therapy(EVLA group),46 cases were treated by transilluminated powered phlebectomy(TIPP group),49 cases were treated by EVLA combined subfascial endoscopic perforator surgery (SEPS)(EVLA+SEPS group) and 54 cases were treated by TIPP combined SEPS (TIPP+SEPS)group.The operation time,the intraoperative blood loss,the postoperative hospitalization cost,the hospitalization time,the incidence rates of postoperative complications (residual varicose veins,subcutaneous induration,superficial phlebitis,skin necrosis and saphenous nerve injury) of the patients in various groups were compared. Results: There were significant differences in the operation time,the intraoperative blood loss and the postoperative hospitalization time,the hospitalization cost of the patients between various groups(P<0.05).Compared with EVLA group,the incidence rats of residual varicose veins,superficial phlebitis,and lower extremity swelling and ecchymosis of the patients in TIPP group were significantly decreased (P<0.05); the incidence rates of subcutaneous induration,wound hematoma,saphenous nerve injury and skin numbness were increased,but there were no significant differences (P>0.05).Compared with EVLA group and TIPP group,the incidence rates of reidual varicose veins and superficial phlebitis in EVLA + SEPS group and TIPP + SEPS group were significantly decreased (P<0.05).The healing rates of the patients 3 months after operation in TIPP group,EVLA + SEPS group and TIPP + SEPS group were significantly increased compared with EVLA group(P<0.05).The recurrence rates 1 year after operation in TIPP group,EVLA + SEPS group and TIPP + SEPS group were lower than that in EVLA group (P<0.05). Conclusion: The curative effects of EVLA combined with SEPS and TIPP combined with SEPS in treatment of varicose veins in lower extremities are superior to EVLA and TIPP,with the advantages of safe and reliable methods,complete varicose vein resection,less postoperative complications,quick ulcer healing and low recurrence rate and so on.

Objective:To investigate the application of non-intravenous dexmedetomidine(DEX) in the pediatric patients underwent lower abdomen and limb surgery, and to observe the sedative effect of DEX in this procedure. Methods: Sixty patients undergoing the general anesthesia for lower abdomen and limb surgery were selected and randomly devided into ropivacaine sacral block (RS) group, intranasal DEX + ropivacaine sacral block (ID) group, ropivacaine + DEX sacral block (DS) group, 20 cases in each group. The children in ID group received intranasal DEX 1 μg·kg^-1 30 min before operation and the children in RS and DS groups received physiological saline. 1 mL·kg^-1 propofol was infused intravenously in the children who could not smoothly enter into the operating room as well as the intolerance to oxygen mask or sevoflurane inhalation while induction. The children in RS and ID groups received 0.25% ropivacaine 1 mL·kg^-1, and the children in DS group received the same dose of ropivacaine mixed with 1 μg·kg^-1 DEX, and the total volume of drugs was 20 mL. The general information of each child was recorded; the sedation status when separated from their parents and induction period mask and sevoflurane acceptance scores were assessed; the satisfaction of separation with parents, oxygen mask and sevoflurane inhalation were recorded; the time of operation, induction, extraction of laryngeal mask and anesthesia awake were recorded; delayed awakening,laryngismus and awakening period agitation score were recorded. The scores of anesthesia recovery and the dosage of propofol were recorded; the sedation scores 4, 8, 12, 16, 20, and 24 h after operation were recorded. Results: Compared with RS and DS groups, the sedation scores of the children when they were separated from their parents and mask induction and sevoflurane inhalation acceptance, the satisfaction degree of separation, mask and sevoflurane acceptance in ID group were increased(P<0.05); the dosage of propofol in ID group were decreased(P<0.05). The time of operation, extraction of laryngeal mask and anesthesia awake had no significant differences between three groups (P>0.05), the induction time of children in ID group was shorter than those in RS and DS groups(P<0.05). There was no delayed awakening in three groups, and the laryngismus and the awakening period agitation score in RS group were higher than those in ID and DS groups (P<0.05). There was no differences in the consciousness, respiration, activity scores and the scores of anethesia recovery between three groups(P>0.05). The sedation scores in the three groups were less than 3 points 4 h after operation. Compared with RS group, the sedation scores in ID and DS groups were decreased 8 h after operation (P<0.05). Compared with RS and ID groups, the sedation scores in DS group 12, 16 and 20h after operation were decreased(P<0.05). There were no significant differences in the sedation scores between three groups 24 h after operation (P>0.05). Conclusion: When non-intravenous DEX is used in the pediatric patients underwent lower abdomen and limb surgery, the children can quietly and co-operationly enter into the operating room and quickly and smoothly complete the induction process; the incidence of revival restlessness is significantly reduced, and it can play a role in the early postoperative sedation.

Objective:To explore the key points of preoperative preparation,the experience of operation and attentions of postoperative treatment of surgical treatment in multiple tophi in the extremities. Methods: A total of 23 cases of multiple tophi in extremities were selected,all were male;aged from 31 to 65 years old, average 48.7 years old;the course of the disease ranged from 4 to 20 years, with an average of 9.13 years;the blood uric acid before operation ranged from 380 to 665 μmol·L^-1,the level of uric acid was controlled by physicians before operation and the patients were not in the attack period. The patients underwent gout stone resection. Some of the patients underwent functional reconstruction and were treated with standardized gout medicine after operation. Results: Twenty-one cases of incision were grade A healing 2 weeks after operation,and 2 cases of incision still had uric acid salt exudation and healed after debridement suture. Among 23 cases,18 patients were followed for 4-12 months, an average of 8 months;5 patients were lost to follow-up. Compared with before operation, the joint function was improved, the degrees of joint activity were increased, and the discomfort was relieved in 16 patients after operation; there was no improvement in postoperative joint function in 2 patients, and the pain symptom in 1 patient with carpal tunnel syndrome was reduced. Conclusion: When properly indicated,the comprehensive therapy is the best treatment for the patients with multiple tophi in extremities.It is reliable and can improve the quality of life of the patients.

Objective:To discuss the clinical characteristics, diagnosis, treatment experience and prognosis of mature cystic teratoma complicated with primary ovarian carcinoid,and to summarize its general charateriseics and to enhance the knowledge of clinician. Methods: Three female patients presented unilateral ovarian tumor in physical examination and were admitted to hospital.The tumor markers of three patients were normal. The preoperative diagnosis was ovarian tumor.Unilateral salpingo-oophorectomy was conducted in two patients.The resection of ovarian tumor was conducted in one patient. Results: The three patients underwent operation successfully.The postoperative pathological diagnosis was mature cystic teratoma complicated with primary ovarian carcinoid.The patients recovered well, who accepted 15-month follow-up with normal life and stable condition,and had no recurrence and metastasis. Conclusion: Mature cystic teratoma complicated with primary ovarian carcinoid is easy to be misdiagnosed before operation.Salpingo-oophorectomy is commonly used as the treatment method of mature cystic teratoma complicated with primary ovarian carcinoid and it can result in good effectiveness and better prognosis.

Objective:To report one case of bilateral maxillary fourth molars, and to explore the occurrence and treatment method of additional fourth molar. Methods: The case materials of one patient with bilateral maxillary four molars were collected, and the information was record. Combined with reviewing the relevant literatures, the clinical data, complication and treatment of one patient with bilateral maxillary fourth molars were retrospectively analyzed. Results: The patient was diagnosed due to left maxillary posterior teeth area intermittent pain for several months and the increased pain for 5 d; at the same time,there was no abnormal appearance inducing the symptoms detected by speciality check-up. The pantomography showed the four molars in bilateral maxillary, and the fourth molars impacted the third molar root's absorption. The fourth molar crown was smaller and the root was shorter. The 18,28,29 teeth were removed,and the left maxillary posterior teeth area intermittent pain disppeared. Conclusion: The occurrence of additional fourth molar is a relatively rare demal dysplasia. However,there are some complications following with the fourth molar. Early diagnosis and treatment are essential.

Objective:To explore the etiology of chylothorax by analyzing the clinical data of one patient with chylothorax and combining with the review of relative literatures, and to improve the ability of clinicians for understanding the rare etiology of chylothorax. Methods: A young female with cough, chest pain and dyspnea as the first symptom was admitted to the hospital. Left lymph node enlargement in the neck of the patient was found in examination and bilateral pleural effusion was found in chest CT scanning. After puncture drainage and testing,the pleural effusion was confirmed the chylothorax.Since the examinations for common causes of chylothorax produced negative results, the patient was eventually diagnosed as infectious chylothorax. The patient received anti-infective therapy for 5 d. Results: After treatment, the symptoms of the patients were improved obviously; the pleural effusion disappeared, and the lymph nodes in the neck were shrink detected with chest color ultrasound. One month after follow-up, all the symptoms, enlarged lymph nodes and pleural effusion were recovered. There was no abnormity in chest CT. Conclusion: The possibility of infection should be considered in clinic for the patients with atypical chylothorax and anti-infective treatment should be performed in order to avoid the failed diagnosis and misdiagnosis.

Objective:To report the clinical manifestations of 1 case of intracranial multicentric astrocytoma, and to provide a reference for its clinical diagnosis and treatment. Methods: The clinical data of one patient with intracranial multicentric astrocytomas were retrospectively analyzed and the diagnosis and treatment were summarized, and the relevant literatures were reviewed. Results: The patient was male, 25 years old,and admitted to hospital due to the sudden convulsions 1 time 18 d ago. The head MRI scanning and enhanced scanning displayed that the left frontal lobe and corpus callosum knee exited the group of patchy mixed abnormal signals, and the left frontal temporal lobe exited the capsule-like mixed signals. Surgical treatment was performed, and the lesions of the left frontal lobe and corpus callosum knee and left frontal temporal lobe were resected. The difference of the two lesions was observed during the operation (the left frontal and corpus callosum knee lesion was cystic and solid mixed tissue, solid organization accounted for majority, and had internal calcification; the left frontal temporal lobe lesion was cystic and solid mixed tissue, and cystic tissue accounted for the majority). The postoperative pathology showed that the left frontal lobe and corpus callosum knee lesion had locally more intensive cells, and more different cells, and it was diagnosed as astrocytoma (WHO Ⅱ-Ⅲ); the left frontal temporal lobe lesion was diagnosed as astrocytoma (WHO Ⅱ). After operation, the patient recovered well, and it was recommended to continue radiotherapy and chemotherapy. Conclusion: For the patients with intracranial multicentric astrocytomas, active surgical treatment is in favor of prolonging the survival of the patients. Postoperative radiotherapy is still controversial, but chemotherapy should be recommended.

Objective:To analyze the clinical pathological features of one patient with accessory breast cancer(ABC),and to explore the diagnosis,treatment, operation methods and prognosis of ABC patient. Methods: The patient received right axillary tumor resection,right axillary accessory breast resection and axillary lymph node dissection, didn't receive resection of breast in the affected side. According to the intraoperative frozen pathological diagnosis, the clinical diagnosis was ABC. After operation, the patient was treated with 8 cycles of AC-T regimen adjuvant chemotherapy(The first four cycles were given pirarubicin 60 mg·m^-2, cyclophosphamide 600 mg·m^-2 per cycle; the last four cycles were given docetaxel 100 mg·m^-2 per cycle; every three weeks was a cycle of treatment), radiation therapy(The radiation dose was 50 Gy/25 f in the upper and lower part of the right collarbone and the tumor bed area, and after retract the tumor bed area was increased to 60 Gy)and endocrine therapy(Tamoxifen was administered at 20 mg per day). Results: The patient's breast color ultrasound and mammogram examination indicated that the right axillary mass of the patient was more likely to be malignant. The clinical diagnosis was right axillary ABC. According to the NCCN guide, the patient was treated with the standardized comprehensive treatment based on surgical treatment. 16 months after operation, the patient recovered well and had a normal life. There was no upper limb dysfunction and no lateral upper limb lymphedema, and there were no recurrence or metastasis. Conclusion: ABC is extremely rarely seen in clinical practice. The clinical pathological features and treatment of ABC are similar to breast cancer. If there is no lesion in the mammary gland, it is not necessary to remove the mammary gland in the affected side.

Objective:To explore the clinical characteristics, the diagnostic framework, and the treatment methods of B cell lymphoblastic lymphoma (B-LBL), and to clarify the progress of diagnosis and treatment of B-LBL to improve the clinician's understanding of the disease and provide the guidance for prognostic evaluation and therapeutic options. Methods: The clinical data including symptoms, physical signs, ancillary testings,diagnosis, treatment and disease prognosis of a child suffered from B-LBL were retrospectively analyzed; in the meantime, the relative literatures were reviewed. Results: The patient was definitly diagnosed as B-LBL according to the clinical characteristics and received combination therapy with vincristine, daunorubicin, L-asparaginase, and prednisone as the first course, along with the intrathecal injection of methotrexate and dexamethasone to prevent central nervous system leukemia (CNS-L). The patient achieved complete remission (CR) 25 d after the first circle chemotherapy but was diagnosed as degree 4 myelosuppression. Therefore, the second cycle combination therapy was adjusted with cyclophosphamide, cytarabine and 6-MP, and the intrathecal injection to prevent CNS concomitantly. Degree Ⅳ myelosuppression appeared repeatedly after 2 cycles and the combination chemotherapy was reajdusted. So mercaptopurine and high dose of methotrexate were given as the 4th cycle,and CNS was prevented continously. The patient kept CR until the second cycle finished but get recurrence after the third chemotherapy(prolymphocytes 10%). Then remission and recurrence were found in the disease counrse during which mary chemotherapy methods were attempted until the patient got stable CR after treatment for 31 months. Then the patient was treated with oral mercaptopurine (50 g·d^-1) and methotrexate (25 mg per week) and kept disease-free survival for more than 3 years. Conclusion: B-LBL is a rapidly developed disease with the bone marrow involvement occurring in the short term and easy to relapse during treatment. However, it is extremely easy to transform to recurrent and refractory B-LBL after the first remission. It is of great importance to estimate the risk stratification and to evaluate the prognosis of LBL patients in order to treat as soon as possible for the improvement of one's life quality and the prolongation of survival.

Objective:To investigate the treatment and aesthetic restoration of anterior tooth loosening and falling in the patient with severe periodontitis, and to clarify the curative effect of multidisciplinary treatment in the anterior esthetic zone, and to provide reference for its clinical application. Methods: The male patients was 57 years old,and saw a dcotor becasuse of anterior teeth loosening and discomfort for more than half a year. The specialized examination:11, 21, 22, 41 Ⅲ° loose, 42 Ⅱ° loose. After multidisciplinary consultation, the teeth were treated with periodontal therapy. After inflammation was controlled, 11, 21, 22, 41, 42 were removed for immediate implantation. The platelet rich fibrin (PRF) combined with guided bone regeneration (GBR) was used to repair the bone defect around the implants. Because of the aesthetic requirements of the patients, tooth preparation was performed for 12, 13, 23, 31, 32, 33, 43 after root canal therapy. Fixed restorations were performed in 13-23 and 33-43. Results: After operation, the implant was well positioned and the bone surrounding the implant was adequate. The soft tissue in the mouth healed well and the degree of the alveolar ridge was better. After restoration, the masticatory and phonation function of the teeth were restored, and the patients were satisfied with the aesthetic effect. Conclusion: Through the application of multidisciplinary treatment, the implant repair of the anterior esthetic zone of the patient with severe periodontitis is successfully achieved, which opens a new way for the diagnosis and treatment of the anterior esthetic zone.

Objective:To evaluate and compare the diagnostic values between myocardial perfusion imaging (MPI) and dual source CT coronary angiography (DS-CTCA) in coronary artery stenosis in the diabetic patients with coronary heart disease (CHD), and to elucidate the clinical application values of MPI combined with DS-CTA in the diabetic patients with CHD. Methods: A total of 52 diabetic patients with CHD underwent the examinations of MPI and DS-CTCA were selected. The degrees of coronary artery stenosis of branches of coronary artery were compared between MPI examination and DS-CTCA examination, including left main coronary artery (LM), left anterior descending coronary artery (LAD), left circumflex branch (LCX) and right coronary artery (RCA). Based on the coronary angiography (CAG) regarded as the golden standard, the specificity, sensitivity and accuracy of each imaging examination method were compared; the specificities, sensitivities and accuracies of parallel diagnosis and serial diagnosis of two kinds of methods were also compared. Results: The stenosis degrees of LM and LAD were no different between MPI examination and DS-CTCA examination (P>0.05), but the stenosis degrees of LCX and RCA were different between two imaging examination methods (P<0.05). Compared with MPI examination, the sensitivity of DS-CTCA examination in the diabetic patients with CHD was decreased (71.0% vs 90.1%), and the difference was significant (P=0.035); the specificity of DS-CTCA examination in the diabetic patients with CHD was increased (85.7% vs 58.4%), and the difference was significant (P=0.027); but the accuracies of two examinations in the diabetic patients with CHD were no different (76.4% vs 78.4%) (P=0.062).Compared with parallel diagnosis, the specificity and accuracy of serial diagnosis were increased(93.5% vs 33.8%, P=0.001; 94.7% vs 71.2%, P=0.030); but the sensitivities had no difference (95.4% vs 93.1%,P=0.074). Conclusion: The diagnostic accuracy of evaluating the degree of coronary artery stenosis in the diabetic patients with CHD is not different between DS-CTCA examination and MPI examination. And the serial diagnosis of two examination methods can improve the diagnostic accuracy.

Objective:To explore the status,the risk factors and prevention strategies and measures of cognitive dysfunction in the patients with type 2 diabetes mellitus (T2DM), and the provide the theoretical basis for preventing the mild cognitive impairment (MCI) in the patients with diabetes mellitus. Methods: A total of 192 patients with T2DM who were diagnosed definitely and received hospitalization treatment and accepted the questionnaire willingly were selected as the subjects. The general demographic characteristics, general behavioral characteristics and clinical data were collected. Montreal Cognitive Assessment (MoCA) was used to screen the status of the patients with cognitive impairment, and Activities of Daily Living (ADL) assessment was used to assess the ADL of the patients. Chi-square test, t test and Wilcoxon test were performed to compare the congnitive function between the patients with different features. Multivariable Logistic regression model was used to find the independent influencing factors of MCI in the patients with T2DM. The correlation analysis was used to analyze the relationships between the each module of cognitive function of MoCA and the influencing factors. The P<0.05 was regarded as a statistically significant value. Results: There were 192 patients with T2DM accepted the questionnaire. According to the criteria for cognitive impairment in MoCA, 154 of the 192 subjects had MCI, and the incidence of MCI was 80.21% (154/192). Multivariate Logistic regression analysis showed that the elderly and low family per capita monthly income were the risk factors of MCI in the patients with T2DM (P<0.05). The results of correlation analysis between the various modules of MoCA and the influencing factors showed that the age was related strongly to the attention ability (r=-0.334, P<0.05); the family per capita monthly income was strongly related to the visual space ability and executive ability (r=0.322, P<0.05). Conclusion: The incidence of MCI in the patients with T2DM is serious. Age and the per capita monthly income of family are the important factors affecting the cognitive function of the patients with T2DM, which mainly affect the visual space ability, executive ability, attention ability, and memory function.

Objective:To prepare the sustained release system of icariin(ICA)@gelatin nanoparticles(GNPs)-polyactic-co-glycolic acid(PLGA) (ICA@GNPs-PLGA), and to optimize the conditions. Methods: ICA@GNPs-PLGA sustained release system was prepared using two-step desolvation method and S/O/W emulsion solvent-evaporation technique. The effects of different conditions, such as the PLGA:GNPs mass ratio and the total quality of ICA added on the entrapment efficiency (EE) of ICA@GNPs-PLGA composite microspheres were detected to optimize the preparation process. The surface morphology of GNPs and ICA@GNPs-PLGA composite microspheres were observed by SEM. The EE and the release results of ICA@GNPs-PLGA in the sample were determined with HPLC. Results: The prepared composite microspheres and nanocomplex were were white powder. The SEM results showed that the composite microspheres and nanocomplexs were spherical, the surfaces were smoothy, and the particle size distribution range was 4-12 μm and 150-200 nm, respectively, relatively uniform. At a GNPs mass fraction of 6 mg, the critical concentration of PLGA in DCM ranged within 0.5%-1.0%. At a GNPs mass fraction of 12 mg, the critical concentration of PLGA in DCM ranged within 1.0%-2.0%.However, at a critical PLGA mass fraction lower than 0.25%, no fully formed composite microspheres were observed. Within the critical concentration, the average EE of ICA@GNPs-PLGA microspheres was higher than(62.00±1.25)%. In addition, the EE of ICA in the microspheres was negatively correlated with the quality of ICA added. The accumulative release rate was less than in 24 h and it was 65.21% in 40 d. Conclusion: The ICA@GNPs-PLGA microspheres with homogeneous particle size distribution, high EE, low initial burst and without agglomeration can be acquired under the optimized conditions.