Objective:To explore the protective effect of glutamine(GLN) on the hyperoxia lung injury in the neonatal rats, and to elucidate its mechanism. Methods:A total of 90 male and female Wistar rats were selected and randomly divided into control group (FiO₂=21%), hyperoxia group(FiO₂> 85%) and hyperioxia + glutamine (GLN) group (FiO₂> 85%)(n=30).The rats in hyperioxia group and hyperioxia+GLN groups were used to establish the models of hyperoxia lung injury(HALI).The rats in hyperoxia+ GLN group were intraperitoneally injected with 0.75 g·kg^-1·d^-1 GLN from the first day of experiment, and the rats in other two groups were abdominally injected with the same volume of normal saline. The body weights, water contents in the lung tissue of the neonatal rats in various groups on the 3rd, 7th, and 14th days of the experiment were measured. HE staining was used to determine the morphology of lung tissue of the rats in various groups; ELISA was used to detect the levels of interleukin-6(IL-6), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the lung tissue homogenate of the rats in various groups. Results:Compared with control group at the same time, the weights of the neonatal rats in hyperoxia group were significantly decreased on the 3rd, 7th and 14th days(P<0.05); the body weights of neonatal rats in hyperoxia + GLN group were significantly higher than those in hyperoxia group at the same time (P<0.05). On the 3rd, 7th, and 14th days, the water contents of lung tissue of the rats in hyperoxia group were higher than those in control group at the same time(P<0.05), and the difference was gradually increased with the prolongation of time; the water contents of lung tissue of the rats in hyperoxia + GLN group were significantly lower than those in hyperoxia group(P<0.05) on the 3rd,7th, and 14th days. Compared with control group, there was a certain degree of damage of the alveolar structure of the rats in hyperoxia group,accompaying with the alveolar fusion, the interval widening, the fibrous tissue hyperplasia and the interstitial inflammatory exudation;the injury degree was aggravated with the prolongation of time. Compared with hyperoxia group, the degrees of alveolar injury, inflammatory exudation, fibrous tissue hyperplasia of the rats in hyperoxia + GLN group were alleviated. Compared with control group at the same time, the levels of IL-1β, IL-6 and TNF-α in lung tissue homogenate of the rats in hyperoxia group were significantly increased (P<0.05);compared with hyperoxia group, the levels of IL-1β, IL-6, and TNF-α of the rats in hyperoxia + GLN group were significantly decreased(P<0.05). Conclusion:GLN has a protective effect on the neonatal rats with HALI, which can alleviate the hyperoxia-induced pulmonary edema in the neonatal rats and inhibit the expressions of IL-6, IL-1β, and TNF-α in lung tissue.

Objective:To observe the regulatory effect of fructus corni on the retinol transport, and to elucidate the mechanism of fructus corni in the treatment of dry eye syndrome. Methods:A totle of 60 BALB/c mice were randomly divided into blank control group, model group, positive control group, low dose of fructus corni group, medium dose of fructus corni group, and high dose of fructus corni group. Benzalkonium chloride eye drop method was used to set up the dry eye syndrome models of the mice.The mice in positive control group were orally administered with Shihu Yeguang Pill(0.2 mL·10g^-1 body mass);the mice in low,medium,and high doses of fructus corni groups were orally administered with 600,1 200 and 2 400 g·L^-1 fructus corni water decoction,respectively(0.2 mL·10 g^-1 body mass);the mice in model group were orally administered with the same volume of normal saline;the mice in various adminstration groups were administered once a day,and lasted for four weeks. The tear secretion amount and tear film rupture time of the mice in vavious groups were detected by tear test paper method and fluoresein staining method.The levels of serum prealbumin(PA),retinol binding protein(RBP) and the retinol in liver tissue of the mice in various groups were detected by ELISA;the expression levels of cellular retinol binding protein (CRBP) and RBP receptor-stimulated by retinoic acid gene 6(STRA6) in lacrimal gland tissue of the mice in various groups were detected by immunohistochemistry and Western blotting methods. Results:Compared with blank control group, the tear secretion amount and the tear film rupture time of the mice in model group were significantly decreased(P<0.01).Compared with model group,the tear secretion amount and the tear film repture time of the mice in positive control group and different doses of fructus corni groups were significnatly increased(P<0.05 or P<0.01). Compared with blank control group, the levels of serum PA and RBP and the levels of retinol in liver of the mice in model group were significantly decreased (P<0.01); the expression levels of CRBP and STRA6 proteins in lacrimal gland tissue were significantly increased(P<0.05 or P<0.01). Compared with model group, the levels of serum PA and RBP and the levels of retinol in liver tissue of the mice in positive control and different doses of fructus corni groups were significantly increased(P<0.01). Compared with positive control group, the levels of serum PA and RBP of the mice in high dose of fructus corni group were increased (P<0.05 or P<0.01); the expression levels of CRBP and STRA6 proteins in lacrimal gland tissue of the mice were significantly increased (P<0.01). Conclusion:When dry eye syndrome occurs,the body is accompanied by deficiency of retinol and obstacle of retinol transport.The sour drug fructus corni can improve the deficiency of retinol by increasing the absorption of retinol,and then regulate the process of retinol transport.

Objective:To explore the expression charateristics of long non-coding RNA bladder cancer associated transcript-1(BLACAT1) in the cancer tissue and cancer cells in the patients with non-small cell lung cancer(NSCLC) and the regulation mechanism,and to elucidate the effect and clinical significance of BLACAT1 in the occurrence and development of NSCLC. Methods:Gene Expression Omnibus (GEO) database was used to analyze the expression characteristics of BLACAT1 in the NSCLC tissue. Kmplot website was used to analyze the correlations between the expression of BLACAT1 and the survival and prognosis of the NSCLC patients. Real-time quantitative PCR(qRT-PCR) method was applied to detect the expressions of BLACAT1 in cancer tissue and corresponding adjacent normal tissue and NSCLC cell lines of the NSCLC patients.The specific small interfering RNA for BLACAT1(si-BLACAT1 group) or negative control sequence(si-NC group) were transfected into the A549 cells. The mRNA expression level of BLACAT1 in A549 cells in various groups were detected by qRT-PCR method; the percentages of A549 cells in different cell cycles were detected by flow cytometry; the clone formation abilities of A549 cells in various groups were detected by cell clone formation experiment. The proliferation activities of cells in various groups were detected by CCK8 assay. qRT-PCR and Western blotting methods were used to detect the expression level of CyclinD1 and CDKN2B mRNA and protein in the A549 cells in various groups. Results:The results of GSE18842 and GSE19804 in GEO datatase,qRT-PCR and Kmplot analysis showed that the expression level of BLACAT1 in NSCLC tissue was significantly increased compared with adjacent normal lung tissue (P< 0.05); the survival time in the patients with low expression of BLACAT1 in cancer tissue was significantly longer than that in the patients with high expression of BLACAT1 (P=0.011). Compared with si-NC group,the BLACAT1 expression level in the A549 cells in si-BLACAT1 group was significantly decreased(P<0.05).Compared with si-NC group, the percentage of A459 cells in G₁ phase in si-BLACAT1 group was increased (P<0.05), and the percentage of A549 cells in S phase was decreased(P<0.05);the cell clone formation ability and the cell proliferation activity were decreased(P<0.05 or P<0.01). Compared with si-NC group, the expression levels of CyclinD1 mRNA and protein in the A549 cells in si-BLACAT1 group were significantly decreased(P<0.05), and the expression levels of CDKN2B mRNA and protein were significantly increased(P<0.05). Conclusion:LncRNA-BLACAT1 is up-regulated in cancer tissue and cancer cells of the NSCLC patients, and down-regulation of BLACAT1 expression can inhibit the proliferation of A549 cellsvia modulating the CyclinD1/CDKN2B axis, which may serve as a potential therapeutic target for the NSCLC patients.

Objective:To construct the recombinant lentiviral vector containing cardiac adriamycin responsive protein (CARP) gene and small-hairpin RNA (shRNA) targeting CARP gene and to pack the lentivirus, and to lay the foundation for further study on the function and mechanism of CARP in adriamycin(ADR)-induced cardiomyopathy. Methods:After the rat CARP gene was amplified by PCR and shRNAs targeting CARP were designed and synthesized, they were inserted into the shuttle plasmids GV-358 or GV-248, respectively. After confirmed by sequencing, the recombinant shuttle vectors containing CARP gene or shRNA and auxiliary packaging plasmid Helper 1.0 and Helper 2.0 were co-transfected into the HEK293T cells for virus packaging and amplification; the viral titer was detected by end-point dilution.The HEK293T cells were infected with the recombinant lentiviruses,and the green fluoresence intensity was observed by fluorescence mircroscope. The H9C2 cells were infected with the recombinant lentivirues and divided into control group, CARP overexpression group and shRNA group; Western blotting method was used to detect the CARP expression levels in the cells in various groups. Results:The DNA sequencing results showed that the sequences of CARP overexpression and shRNA vectors were in accordance with the designed sequences. The expression of green fluorescence protein was seen under fluorescence microscope after transfection of the vectors of the HEK293T cells. After infection of the H9C2 cells, the expression level of CARP protein in CARP overexpression group was 5.3 times higher than that in control group (P=0.01); while it was down-regulated by 53% in CARP shRNA group compared with control group (P=0.02). Conclusion:The lentivirus expression vectors carrying CARP or shRNA targeting CARP are successfully constructed and the lentiviruses obtained could significantly interfere the expression of CARP in the H9C2 cells.

Objective:To observe the influence of miRNA-21 in the radiosensitivity of the breast cancer cells, and to preliminarily explore its mechanism. Methods:The breast cancer T47D and MDA-MB-361 cells were irradiated with 0.0,2.5,and 5.0 Gy γ-ray,respectively; CCK8 assay was used to detect the survival rates of T47D and MDA-MB-361 cells and flow cytometry was used to analyze the cell cycle; real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells during 72 h. The T47D and MDA-MB-361 cells were transfected with anti-miRNA-21 sequence (anti-miRNA-21 group) and negative control sequence(negative control group), respectively; the untransfected cells were set as blank control group. Real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells in various groups. After irradiation with 0.0 and 5.0 Gy γ-ray, CCK8 assay was used to detect the survival rates, and flow cytometry was used to analyze the cell cycle. Results:After irradiation with 5.0 Gy γ-ray, the survival rate of T47D cells was significantly higher than that of the MDA-MB-361 cells (P<0.05). Compared with 0.0 Gy radiation group, the percentages of T47D and MDA-MB-361 cells in G₂/M phase in 5.0 Gy radiation group were significantly increased(P<0.05); the miRNA-21 expression levels in the T47D and MDA-MB-361 cells were significantly increased (P<0.05).The miRNA-21 expression levels in the T47D and MDA-MB-361 cells in anti-miRNA-21 group were lower than those in blank control group (P<0.05). After irradiation with 5.0 Gy γ-ray, the survival rates of T47D and MDA-MB-361 cells in anti-miRNA-21 group were significantly lower than those in blank control group(P<0.05 or P<0.01), and the percentage of T47D cells in G₂/M phase in anti-miRNA-21 group was significantly lower than that in blank control group (P<0.05), and the percentage of T47D cells in subG₁ phase was significantly higher than that in blank control group (P<0.05). Conclusion:The down-regulation of miRNA-21 may enhance the radiosensitivity of breast cancer cells, and its mechanism may be related to the inhibition of G₂/M phase arrest.

Objective:To investigate the effect of herba artemisiae capillaris extracts (HACE) on the expression of phosphatase and tensin homologue deleted chromatosome 10 (PTEN) in the kidney tissue of the diabetic rats, and to clarify the pharmacological mechanism of protective effect of HACE on the kidney of the diabetic rats. Methods:Thirty Wistar rats were randomly divided into control group (n=6), model group (n=12) and HACE group (n=12).The rats in HACE group were intragastrically administrated with HACE(5 g·kg^-1) one time every day,and the rats in control group and model group were intragastrically administrated with the same volume of normal saline.The diabetic rat models were duplicated by streptozotocin (STZ). After 4 months of administration, the urinary albumin excretion rates and urinary total protein excretion rates of the rats in various groups were detected. The pathomorphology of the kindey tissue of the rats in various groups were observed by HE staining and the distribution of renal extracellular matrix (ECM) was detected by PAS and Masson staining. The expressions of PTEN protein in kidney tissue of the rats in various groups were detected by immunohistostaining and the expression levels of PTEN protein in kidney tissue of the rats in various groups were detected by Western blotting method. Results:Compared with model group, the glomerular mesangial aggregation, positive staining of PAS as well as Masson aggregation and basement membrane thickening of the rats in HACE group were significantly reduced, and the 24 h urinary albumin excretion rate was significantly decreased (P<0.05),and the urinary total protein excretion rate had no significant difference(P>0.05).The immunohistochemistry staining results showed that the expression level of PTEN protein in kidney tissue of the rats in control group was higher, the expression level of PTEN protein in the kidney tissue of the rats in model group was decreased, and the expression level of PTEN protein in kidney tissue of the rats in HACE group was increased to normal. The Western blotting results showed that the expression level of PTEN protein in the renal cortex tissue tissue of the rats in model group was significantly decreased compared with control group (P<0.05); compared with model group, the expression level of PTEN protein in renal cortex tissue of the rats in HACE group was significantly increased (P<0.05). Conclusion:HACE can increase the expression of PTEN protein in kidney tissue of the rats, which may be one of the mechanisms of its protective effect on the kidney in the diabetic rats.

Objective:To investigate the effects of oxaliplatin combined with capecitabine on the expressions of tumor-related factors in the gastric cancer tissue and the levels of serum inflammatory factors in the rats with experimental gastric cancer, and to elucidate the possible mechanism of their anti-tumor effects. Methods:Sixty Wistar rats were randomly divided into control group, model group, oxaliplatin group, capecitabine group and oxaliplatin combined with capecitabine group(combination group);there were 12 rats in each group. The BGC823 cells were used to establish the experimental gastric cancer models. The experimental rats were given capecitabine(400 mg·kg^-1) and(or) oxaliplatin(20 mg·kg^-1) at the same time; the rats in control group and model group were given the normal saline at the same volume for 8 weeks.The average tumor weight was measured after administration. The apoptosis of gastric cancer cells was measured by TUNEL staining. The protein expression levels of P53, signal transduction and activation factor 3(STAT3) and vascular endothelial growth factor(VEGF) in the gastric cancer tissue of the rats in various groups were detected by Western blotting method. The levels of serum interleukin-6(IL-6),interleukin-10(IL-10) and tumor necrosis factor-α(TNF-α) of the rats in various groups were detected by ELISA. Results:Compared with control group, the protein expression levels of P53, STAT3, and VEGF in the gastric cancer tissue and the levels of serum IL-6, IL-10, and TNF-α of the rats in model group were significantly increased (P<0.05). Compared with model group, the average tumor weights and the protein expression levels of P53, STAT3, and VEGF in the gastric cancer tissue and the levels of serum IL-6, IL-10,and TNF-α of the rats in oxaliplatin group, capecitabine group and combination group were significantly decreased (P<0.05); the apoptosis indexes(AI) of gastric cancer cells were significantly increased(P<0.05). The average tumor weight,the protein expression levels of P53, STAT3,and VEGF in gastric cancer tissue and the levels of serum IL-6, IL-10, and TNF-α of the rats in combination group were significantly lower than those in capecitabine group and oxaliplatin group(P<0.05). Conclusion:Both oxaliplatin and capecitabine can paly the anti-tumor effects by reducing the protein expression levels of the tumor-related factors and the serum IL-6, IL-10, and TNF-α levels of the gastric cancer rats, and the anti-tumor effect of combined utilization is better than that of capecitabine or oxaliplatin used alone.

Objective:To investigate the partial delivery ability in vivo of oligonucleotide MT01 mediated by polyethyleneimine(PEI) derivative PEN, and to illuminate its effect on the alveolar bone reconstruction and the biological safety in the rats with experimental tooth movement. Methods:Forty-eight Wistar male rats were randomly divided into PEN,MT01, sulfuration-modified MT01 (MT01s), and PEN/MT01 groups. The rat experimental tooth model was established, and the above four drugs were injected into the mesial buccal mucosa of the first molar in the left sides of the rats as drug intervention sides,and the PBS were injected into the mesial buccal mucosa of the first molar in the right sides as PBS control sides;once every 3 d. After 14 d, the rats were sacrificed. HE staining was used to detect the histopathological changes of important organs of the rats;the photos and X-ray films were applied to measure the distance of tooth movement, and RT-PCR was used to detect the expression levels of the osteogenic marker genes Runt-related transcription factor 2(Runx2),special protein 7(SP7) and osteocalcin(OCN) mRNA. Results:After local injection of the above drugs, no obvious inflammatory cell infiltration and structural differences were found in the main organs of the rats and no obvious abnormalities were found in all organs tissues. Compared with the PBS control sides, the movement distance of the first molars in the drug intervention sides in MT01, MT01s, and PEN/MT01 groups was reduced (P<0.05 or P<0.01), but there was no significant difference in PEN group (P>0.05). The data of the distance between the two teeth of the rats in various groups was PEN/MT01 group > MT01s group > MT01 group>PEN group, and the difference between PEN/MT01 group and MT01s group was statistically significant (P<0.05).The expression levels of Runx2, SP7, and OCN mRNA in the periodontium tissue in the drug intervention sides of the rats in MT01s and PEN/MT01 groups were significantly higher than those in the PBS control sides (P<0.05 or P<0.01), especially in PEN/MT01 group.Compared with the PBS control sides,the expression level of Runx2 mRNA in the periodontium tissue in the drug intervention sides of the rats in MT01 group was decreased(P<0.05), and the expression levels of SP7 and OCN mRNA in the periodontal tissue of the rats in MT01 group were increased(P<0.05);the expression level of Runx2 mRNA in the periodontium tissue in the drug intervention sides of the rats in PEN group was decreased(P<0.05), and there were no significant differences in the expression levels of SP7 and OCN mRNA (P>0.05). Conclusion:PEI derivative PEN can transfer MT01 into the cell safely and efficiently; it can increase the expression levels of Runx2, SP7 and OCN mRNA in the periodontium tissue and decrease the tooth movement distance in the experimental tooth movement rats.

Objective:To investigate the expression of vascular endothelial growth factor (VEGF) during the healing process of tooth extraction, and to clarify the effect of panax notoginseng saponins(PNS) in promoting the healing of extraction and its mechanism. Methods:Forty-five SD male rats were randomly divided into control group(not given any drugs),chitosan thermosensitive gels group(given chitosan thermosensitive gels without panax notoginseng saponins), 0.5 g·L^-1 PNS group(given chitosan thermosensitive gels and 0.5 g·L^-1 PNS),1.0 g·L^-1 PNS group (given chitosan thermosensitive gels and 1.0 g·L^-1 PNS),and 1.5 g·L^-1 PNS group(given chitosan thermosensitive gels and 1.5 g·L^-1 PNS), with 9 rats in each group.After the middle incisors of right lower jaw were removed, about 20 μL of chitosan thermosensitive gels containing different concentrations of PNS was injected into the each tooth extraction wound of the rats in other groups,except control group. The rats were sacrificed at 1, 2, and 4 weeks after administration, and the specimens were prepared.HE staining was used to observe the healing status of tooth extraction wounds at 1, 2, and 4 weeks after administration.Immunohistochemical staining was performed to detect the number of VEGF positive cells and the expression levels of VEGF protein at 1,2, and 4 weeks after administration. Results:The HE staining resuls showed that the healing status of tooth extraction wounds in chitosan thermosensitive gels group was similar to that in control group;compared with control group, the osteoblasts and vascular endothelial cells were highly differentiated and migrated and the number of bone trabecula was increased. Compared with control group, at 1,2, and 4 weeks after administration, there were no significant differences in the number of VEGF positive cells and the protein expression level of VEGF in the tooth extraction wound tissue of the rats in chitosan gel group (P>0.05); the number of VEGF positive cells and the protein expression level of VEGF of the rats in 0.5,1.0,and 1.5 g·L^-1 PNS groups were significantly increased (P<0.05). Conclusion:PNS can up-regulate the expressions of VEGF in vascular endothelial cells and osteoblasts in tooth extraction wounds of the rats, induce the differentiation of vascular endothelial cells and osteoblasts, increase the number of migration and aggregation of cells, and achieve the effect of promoting the tooth extraction wound healing.

Objective:To detect the effect of over-expressed human MIR31HG gene on the proliferation and migration of PC9 cells, and to clarify the mechanism of oncogene MIR31HG. Methods:The full-length sequence of long non-coding RNA (LncRNA) MIR31HG was amplified by RT-PCR and cloned into the pcDNA3.1(-) eukaryotic expression vector. The PC9 cells were transfected with the pcDNA3.1-MIR31HG overexpression vector and control vector pcDNA3.1. The construction of MIR31HG overexpression vector was detected by enzymatic digestion identification. The stable cell lines with overexpression of MIR31HG (PC9-pcDNA3.1-MIR31HG,stable transfection group) and control cell lines (PC9-pcDNA3.1,empty vector group) were established by G418 drug screening, and the expression level of MIR31HG gene in stably transfected cell line was detected by RT-PCR. CCK-8 method and scratch healing assay were used to detect the proliferation activities and migration abilities of PC9 cells. Results:The agarose gel electrophoresis results showed that the specific gene fragment of MIR31HG was obtained by amplification successfully. The gene fragments of target gene and vector were produced by double enzyme digestion of MIR31HG eukaryotic expression vector. The RT-PCR results showed that the MIR31HG RNA expression level in the cells in stable transfection group was significantly higher than that in empty vector group(P<0.05).The results of CCK-8 test showed that the proliferation activities of the cells in stable transfection group were significantly higher than those in empty vector group at 24, 36 and 48 h after culture(P<0.01).The results of scratch healing assay showed that the scratch healing rate of cells in stable transfection group was significantly higher than that in empty vector group at 48 h after culture(P<0.05). Conclusion:The eukaryotic overexpression vector and the PC9 cell line stably transfected with human LncRNA MIR31HG gene are constructed successfully, and MIR31HG overexpression can promote the proliferation and migration of lung cancer PC9 cells.

Objective:To investigate the influence of panax notoginseng saponins (PNS) in the T lymphocyte subsets and hematopoietic function in the aplastic anemia model mice, and to elucidate the therapeutic mechanism of PNS in the aplastic anemia mice. Methods:Fifty mice were randomly divided into control group, model group, low dose of PNS group, medium dose of PNS group, and high dose of PNS group.The mice in low,medium,and high doses of PNS groups were intraperitoneally injected with 100, 200, and 400 mg·kg^-1PNS; the mice in control group and model group were intraperitoneally injected with normal saline of the same volume;lasted for 14 d. The body weights of the mice were measured and the spleen and thymus indexes were calculated after administration. Flow cytometry was used to detect the percentages of CD4⁺, CD8⁺T lymphocytes and the ratio of CD4⁺/CD8⁺T lymphocytes in the peripheral blood of the mice.The serum γ-interferon (INF-γ) and tumor necrosis factor-α(TNF-α) levels of the mice were determined by ELISA. The number of red blood cells (RBC), bone marrow mononuclear cells (BMCs), white blood cells (WBC), platelets (Plt) and the hemoglobin (Hb) level of the mice were measured by hematdogy analyzer. Results:Compared with control group, the body weight, the spleen index,the thymus index, the peripheral blood CD4⁺T lymphocyte percentage and the CD4⁺/CD8⁺T lymphocyte ratio of the mice in model group were significantly decreased (P<0.05); the number of RBC, BMCs, WBC, Pl tand the Hb level were significantly decreased(P<0.05); the peripheral blood CD8⁺T lymphocyte percentage and the serum INF-γ, TNF-α levels were significantly increased (P<0.05). Compared with model group, the body weights, the spleen indexes, the thymus indexes, the CD4⁺T lymphocyte percentages and the CD4⁺/CD8⁺T lymphocyte ratios of the mice in low, medium, and high doses of PNS groups were significantly increased (P<0.05); the number of BMCs, WBC, Plt and the level of Hb were significantly increased(P<0.05); the percentages of CD8⁺T lymphocytes and the levels of serum INF-γ and TNF-α of the mice were significantly decreased (P<0.05). Conclusion:PNS can improve the hematopoietic function of the aplastic anemia mice by regulating the balance of T lymphocyte subsets in a dose-dependent manner and inhibiting the release of INF-γ and TNF-α and increasing the levels of hematopoietic related factors.

Objective:To investigate the effects of Urantide on the expressions of c-Jun N-terminal kinase (JNK) mRNA and protein in the thoracic aorta tissue of the rats with atherosclerosis (AS), and to elucidate the molecular mechanism and significance of prevention and treatment of AS. Methods:The AS rat models were established by intraperitoneal injection of Vitamin D₃ combined with high-fat diet and control group was set up at the same time. A total of 150 AS model rats were divided into model group, simvastatin group, Urantide 3 d group, Urantide 7 d group and Urantide 14 d group(n=30).The rats in simvastatin group were adminstrated with simvastatin by gavage for 14 d,and the rats in Urantide groups were injected with Urantide by caudal vein for 3,7,and 14 d. The serum markers of the rats in various groups were detected by automatic biochemical analyzer; the morphology of rat thoracic aorta tissue was observed by HE staining; the expression levels of JNK mRNA and protein in the rat thoracic aorta tissue were detected by immunohistochemical staining, qRT-PCR and Western blotting methods. Results:Compared with control group, the serum levels of triglyceride (TG), total cholesterol (TC) and low density lipoprotein (LDL) of the rats in model group were increased (P<0.05), and the level of high-density lipoprotein (HDL) was decreased (P<0.05).The HE staining results showed the formation of bubbling cells in the thoracic aorta tissue, rupture of medullary elastic fibers and calcification of the rats in model group;compared with model group, the pathological symptoms of the thoracic aorta tissue of the rats in simvastatin group and Urantide groups were improved. The immunohistochemistry results showed that the JNK positive particles were weakly expressed in the rat thoracic aorta tissue in control group; the expression intensity of JNK in the rat thoracic aorta tissue in model group was increased (P<0.05);compared with model group,the expression intensities of JNK in the rat thoracia aorta tissue in Urantide groups were significantly decreased(P<0.05). The qRT-PCR and Western blotting results showed that the expression levels of JNK mRNA and protein in the rat thoracic aorta tissue in model group were significantly increased(P<0.05) compared with control group; compared with model group,the expression levels of JNK mRNA and protein in simvastatin group and Urantide groups were gradually decreased (P<0.05);compared with simvastatin group,the expression levels of JNK mRNA and protein in the rat thoracic aorta tissue in Urantide groups were significantly decreased(P<0.05). Conclusion:Urantide can improve the pathological damage of the thoracic aorta in the AS rats, and its mechanism may be related to the inhibition of JNK signaling pathway expression.

Objective:To investigate the effect of knockdown of EphB1 gene on the oxidative damage of cardiomyocytes (H9c2) induced by hydrogen peroxide (H₂O₂) in the rats, and to provide the evidence for EphB1 gene in the treatment of cardiovascular diseases. Methods:The rat cardiomyocytes H9c2 were cultivated in vitro and divided into blank group (no treatment), H₂O₂ group (H₂O₂-induced cell damage), negative control group (transfected with siRNA control) and si-EphB1 transfection group (transfected with si-EphB1 after H₂O₂-induced cell damage). The expression levels of EphB1 mRNA in the H9c2 cells in various groups were detected by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR), the survival rates of the H9c2 cells in various groups were tested by MTT assay, the apoptosis of H9c2 cells were measured by flow cytometry, the ROS levels in the H9c2 cells in various groups were determined by DCFH-DA, and the levels of MDA and SOD in the H9c2 cells in various groups were determined by spectrophotometer. Results:Compared with blank group, the expression level of EphB1 mRNA in the H9c2 cells in H₂O₂ group were significantly increased(P<0.05);the expression level of EphB1 mRNA in the H9c2 cells in negative control group had no change(P>0.05); compared with H₂O₂ group,the expression level of EphB1 mRNA in the H9c2 cells in si-EphB1 transfection group was significantly decreased(P<0.05).Compared with blank group,the survival rate of H9c2 cells in H₂O₂ group was decreased(P<0.05), the apoptotic rate was increased(P<0.05), the ROS and MDA levels were increased(P<0.05), and the SOD level was decreased(P<0.05);but the differences in the survival rate, apoptotic rate, ROS, MDA and SOD levels in negative control group were not significant(P>0.05).Compared with H₂O₂ group, the survival rate of H9c2 cells in si-EphB1 transfection group was increased(P<0.05), the apoptotic rate was decreased(P<0.05), the ROS and MDA levels were decreased(P<0.05), and the SOD level was increased(P<0.05). Conclusion:Knockdown of EphB1 gene can significantly inhibit the H₂O₂-induced oxidative damage of the rat cardiomyocytes and protect against myocardial injury; its mechanism is related to improving the survival rate of cardiomyocytes, inhibiting the apoptosis, improving the antioxidant enzyme activity, and increasing the scavenging abilities of oxidative stress products in the cells.

Objective::To observe the expression of divalent metal transporter 1 (DMT1) in neurons under the condition of Ndfip1 overexpression, and to explore the regulation effect of Ndfip1 on DMT1. Methods:The Ndfip1 eukaryotic expression plasmid was constructed, and the human neuroblastoma SH-SY5Y cells were transfected with Ndfip1 plasmid; the transfection efficiency was detected. The SH-SY5Y cells transfected with Ndfip1 plasmid were used as experimental group,and the SH-SY5Y cells transfected with empty plasmid were used as control group. The expression intensities of DMT1 protein in the SH-SY5Y cells in two groups were observed by immunofluorescence method,and the expression levels of DMT1 protein in the SH-SY5Y cells in two groups were detected by Western blotting method. Results:The plasmid was amplified, electrophoretically separated and sequenced, and the results proved that the plasmid was successfully constructed. After transfection of SH-SY5Y cells with Ndfip1 plasmid for 48 h, the expression level of Ndfip1 was significantly increased compared with before transfection (P<0.01). The immunofluorescence results showed that the fluorescence intensity of DMT1 in the cells in experimental group was significantly decreased compared with control group. The Western blotting results showed that the expression level of DMT1 in the cells in experimental group was decreased (P<0.05) compared with control group. Conclusion:The overexpression of Ndfip1 can down-regulate the expression of DMT1 protein in the nerve cells, and Ndfip1 nerve has a negatively regulatory effect.

Objective:To investigate the improvement effect of estradiol valerate combined with aspirin on the intrauterine adhesion (IUA) in the rats, and to elucidate the therapeutic mechanism of estradiol valerate combined with aspirin. Methods:Fifty healthy female SD rats were randomly divided into control group, model group, estradiol valerate group, aspirin group,and estradiol valerate combined with aspirin group (combination group)(n=10);the rats in the other groups except control group were used to establish the IUA models with double injury method of uterine curettage and infection. The serum and uterine tissue were taken from the rats in each group 24 h after continuous administration, the estradiol levels in serum of the rats were observed by ELISA, and the pathological changes and the number of glands in the uterine cavity of the rats in various groups were observed by HE staining, and the ratio of endometrial fibrosis are a was observed by Masson staining. The expression levels of transforming growth factor-β1 (TGF-β1), plasminogen activator inhibitor type-1 (PAI-1) and matrix metalloproteinase-9 (MMP-9) in the endometrium of the rats were detected by immunohistochemistry. Results:The results of ELISA indicated that the estrodiol level in serum of the rats in model group was significantly lower than that in control group(P<0.05);compared with model group,the estrodiol levels in serum of the rats in estradiol valerate group, aspirin group, and combination group were significantly increased (P<0.05). The results of HE and Masson staining showed that compared with control group, uterine cavity adhesion and enlargement, fluid accumulation in the cavity, disorganized arrangement of the cells of uterine cavity wall, significant infiltration of inflammatory cells, endometrial stroma fibrosis,and disordered collagenous fibers were found in model group; the number of endometrial glands in the rats was significantly decreased(P<0.05),and the fibrosis area ratio was significantly increased (P<0.05). Compared with model gorup,the adhesion and effuson in uterine cavity,inflammatory cell infiltration and endometrial stroma fibrosis were alleviated,the cells of uterine cavity wall were arranged neatly,and there was little collagenous fibers in estradiol valerate group,aspirin group,and combination group;the number of endometrial glands was increased(P<0.05),and the fibrosis area ratios were significantlhy decreased(P<0.01).Compared with aspirin group,the number of endometrial glands in combination group was significantly increased(P<0.05),and the fibrosis area ratio was significantly decreased(P<0.05).Compared with model group, the expression levels of TGF-β1 and PAI-1 proteins in the endometrium tissue of the rats in estradiol valerate group, aspirin group, and combination group were significantly decreased (P<0.05 or P<0.01), and the expression levels of MMP-9 protein in the endometrium tissue were significantly increased (P<0.05).The expression levels of TGF-β1 and PAI-1 proteins in the endometrium tissue of the rats in combination group were significantly lower than those in aspirin group (P<0.05), and the expression level of MMP-9 in the endometrium tissue was significantly higher than that in aspirin group (P<0.05). Conclusion:Estradiol valerate combined with aspirin can decrease the expression levels of TGF-β1 and PAI-1 in the endometrium tissue and increase the expression level of NMP-9, which can improve the IUA of the rats.

Objective:To investigate the influence of Prunus tomentosa thunb total flavonoids (PTTTF) in the inflammatory factors in the lipopolysaccharide(LPS)-induced cellular inflammation model, and to clarify its mechanism. Methods:The cellular inflammation models of the RAW264.7 cells and human neutrophils (PMN) induced by LPS were established. The cells were divided into control group, model group, dexamethasone group (DXM), and different doses (0.4,4.0, and 40.0 mg·L^-1) of PTTTF groups. The expression levels of inflammatory factors including interleukin-1β(IL-1β), interleukin-6(IL-6), inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α) and high mobility group protein B1(HMGB1)mRNA in the RAW264.7 cells were detected by RT-PCR. The expression levels of NF-κB, and signal pathway key molecules ERK1/2 and p-JNK proteins in the RAW264.7 cells were determined by Western blotting method. Flow cytometry was used to detect the percentages of RAW264.7 cells in different cell cycles and the apoptotic rates of PMN. Results:Compared with control group, the expression levels of IL-1β, IL-6, iNOS, TNF-α, and HMGB1 mRNA in the RAW264.7 cells in model group were significantly increased (P<0.05 or P<0.01),and the expression levels of NF-κB,ERK1/2, and p-JNK proteins were also significantly increased (P<0.01);the percentage of RAW264.7 cells in S phase was increased (P<0.05);the apoptotic rate of PMN was significantly decreased (P<0.05). Compared with model group,the expression levels of IL-1β, iNOS, TNF-α, and HMGB1 mRNA in the RAW264.7 cells in 4.0 and 40.0 mg·L^-1 PTTTF groups and DXM group were significantly decreased(P<0.05 or P<0.01), the expression levels of NF-κB,ERK1/2, and p-JNK proteins were significantly decreased(P<0.05), and the percentages of RAW264.7 cells in S phase were significantly decreased (P<0.05 or P<0.01); the apoptotic rates of PMN were increased(P<0.05 or P<0.01). Conclusion:PTTTF can down-regulate the expression levels of inflammatory factors in the RAW264.7 cell inflammation models induced by LPS. This effect may be related to decreasing the percentage of inflammatory cells in S phase, promoting the apoptosis, and down-regulating the expressions of NF-κB, ERK1/2, and p-JNK proteins.

Objective:To investigate the protective effect of butylphthalide on the brain of the rats with acute ischemic stroke, and to elucidate its mechanism in the treatment of acute ischemic stroke. Methods:Forty-eight rats were randomly divided into sham operation group, model group (focal cerebral ischemia rat model) and butylphthalide group (focal cerebral ischemia rat model + butylphthalide treatment), with 16 rats in each group.The nerve symptom scores of the rats in various groups were determined, and the brain tissue was stained by HE staining; triphenyl four azole nitrogen chloride (TTC) was used to detect the percentage of cerebral infarction area; Western blotting method and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression levels of hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinases-9 (MMP-9), and nuclear transcription factor inhibiting protein kappa B alpha(IκBα), nuclear transcription factor kappa B (NF-κB) p65(NF-κB p65) protein and mRNA in the brain tissue of the rats. Results:In sham operation group, there were no infarction and neurological defect in the brain tissue. Compared with model group, the percentage of cerebral infarction area and the neurological function score of the rats in butylphthalide group were significantly decreased(P<0.01). The structure and distribution of brain tissue cells of the rats in sham operation group were intact and uniform; most of the cells in model group were necrotic, cytoplasmic rupture, nucleus rupture and condensation; a small number of brain cells in butylphthalide group were swollen and necrotic. Compared with sham operation group, the expression levels of HGF, VEGF, MMP-2, MMP-9, and NF-κB p65 protein and mRNA in the brain tissue of the rats in model group were significantly decreased (P<0.01), and the expression levels of IκBα protein and mRNA were significantly increased (P<0.01). Compared with model group, the expression levels of HGF, VEGF, MMP-2, MMP-9, and NF-κB p65 protein and mRNA in the brain tissue of the rats in butylphthalide group were significantly increased (P<0.01),and the expression levels of IκBα protein and mRNA were significantly decreased (P<0.01). Conclusion:Butylphthalide can improve the nerve function of the rats with acute ischemic stroke and reduce the area of cerebral infarction;its mechanism may be related to increasing the HGF level in the brain tissue and promoting the cerebral angiogenesis.

Objective:To investigate the effects of berberine combined with simvastatin on the expressions of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), interleukin-18 (IL-18) and interleukin-10 (IL-10) in the thoracic aorta tissue of the atherosclerosis (AS)model rats, and to elucidate the mechanism of anti-atherosclerosis of berberine combined with simvastatin. Methods:A total of 48 rats were divided into control group, model group, simvastatin group, low, middle and high doses of berberines combined with simvastatin groups(n=8).The serum and thoracic aorta tissue of the rats were obtained 4 weeks after adminstration. HE staining was used to observe the pathomorphology of the thoracic aorta of the rats in various goups; ABC-double antibody sandwich ELISA method was used to detect the serum VEGF, TGF-β1, IL-18 and IL-10 of the rats in various groups; automatic biochemistry analyzer was performed to determine the levels of serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterin (LDL-C), high density lipoprotein cholesterin (HDL-C) of the rats in various groups. The expression levels of VEGF, TGF-β1, IL-18, and IL-10 proteins in the aortic tissue of the rats in various groups were determined by immunohistochemistry. Results:The HE results showed that compared with control group, the intima of aorta of the rats in model group was significantly thickened, lipid deposition was observed inside and outside the cells, necrosis was observed under the intima, plaques and irregularities were observed in the inner wall, and the vascular cavity was narrowed;compared with model group, in high dose of berberine combined with simvastatin group, there was no obvious plaque on the arterial wall, and the intimal thickening was significantly improved. Compared with model group, the serum TC, TG, and LDL-C levels of the rats in simvastatin group and different doses of berberine combined with simvastatin group were significantly decreased (P< 0.05 or P< 0.01), and the serum HDL-C level was significantly increased (P< 0.05). The results of ABC-ELISA showed that compared with model group, the serum VEGF and IL-18 levels in simvastatin and different doses of berberine combined with simvastatin groups were significantly decreased (P<0.05 or P<0.01),and the serum TGF-β1 and IL-10 levels were significantly increased (P<0.05). The results of immunohistochemistry showed that compared with model group,the expression level of IL-18 protein in thoracic aorta tissue of the rats in simvastatin group was significantly decreased(P<0.05),and the expression level of TGF-β1 protein was significantly increased(P<0.05); the expression levels of VEGF and IL-18 proteins in the thoracic aorta tissue of the rats in different doses of berberine combined with simvastatin groups were significantly reduced (P<0.05),and the expression levels of TGF-β1 and IL-10 proteins were significantly increased (P<0.05). Conclusion:Berberine combined with simvastatin could control AS via the up-regulation of the expression levels of VEGF and IL-18 proteins and the down-regulation of the expression levels of TGF-β1 and IL-10 proteins in serum and thoracic aorta tissue of rats.

Objective:To investigate the effect of bromodomain-containing protein 4(BRD4) gene on the apoptosis of cardiomyocytes induced by transforming growth factor-β1 (TGF-β1), and to clarify its possible mechanism. Methods:The H9c2 cells were randomly divided into blank control group,si-CN group (transfected with negative control siRNA),and si-BRD4 group (transfected with specific siRNA).The expression levels of BRD4 protein in the cells in various groups were detected by Western blotting method.The H9c2 cells were randomly divided into control group, TGF-β1 group (transfected with 20μg·L^-1TGF-β1 for 24 h), si-NC+TGF-β1 group (treated with TGF-β1 for 24 h after transfected with negative control siRNA) and si-BRD4+TGF-β1 group (treated with TGF-β1 for 24 h after transfected with BRD4 specific siRNA).The cell proliferation activity was measured by MTT assay, and the apoptotic rate of cells was detected by Annexin Ⅴ-FITC/PI double staining. Western blotting method was used to detect the expressions of BRD4, Bcl-2, Bax, Cleaved caspase 3, P38, and p-P38 proteins. Results:Compared with blank control group, the expression level of BRD4 protein in the H9c2 cells in si-BRD4 group was significantly decreased (P<0.01).Compared with control group, the cell proferation activity and the Bcl-2 protein expression level in TGF-β1 group were significantly decreased(P<0.01); the apoptotic rate, and the expression levels of Bax, Cleaved caspase3, and p-p38 proteins were significantly increased (P<0.01).Compared with TGF-β1 group, the cell prdiferation activity and the expression level of Bcl-2 protein in si-BRD4+TGF-β1 group were significantly increased (P<0.05), while the apoptotic rate and the expression levels of Bax, Cleaved caspase3, and p-p38 proteins were significantly reduced(P<0.05). Conclusion:Inhibition of BRD4 gene expression can attenuate the apoptosis of cardiomyocytes induced by TGF-β1, which may be related to the inhibition of p38MAPK signaling pathway.

Objective:To construct the form deprivation myopia (FDM) rat models, and to elucidate the expression of transforming growth factor-β1 (TGF-β1) in scleral fibroblasts of the FDM rats and its relationship with Wnt/β-catenin signaling pathway. Methods:Forty rats were randomly divided into control group and FDM model group, with 20 rats in each group.The FDM rat model was established in FDM model group.The axial lengths of the rats were determined.The rat sclera tissue of eyeball was separated.The expression levels of TGF-β1 protein and mRNA in sclera tissue of the rats were determined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) methods.The scleral fibroblasts of rats were isolated and cultured.The fibroblasts in the sclera of the rats in control group were used as the control group.The fibroblasts in FDM model group were divided into FDM group and FDM+Dickkopf related protein 1 (DDK1) group (added to DDK1 to culture).The expression levels of TGF-β1, Dicer-like 3 (DCL3), colon adenomatous polyp protein (APC), glycogen synthase kinase 3β (GSK3β), p21-GSK3β, and β-catenin protein and mRNA in scleral fibroblasts were determined by Western blotting and RT-PCR methods. Results:Compared with control group,the axial length of the rats in FDM group was increased (P<0.01);the expression levels of TGF-β1 protein and mRNA in sclera tissue were significantly decreased(P<0.01).There were no significant differences in the expression levels of GSK3β protein and mRNA in scleral fibroblasts between control group,FDM group and FDM+DDK1 group (P>0.05);but there were significant differences in the expression levels of TGF-β1,DCL3,APC,p21-GSK3β,and β-catenin protein and mRNA in the scleral fibroblasts(P<0.01).Compared with control group, the expression levels of TGF-β1 and APC protein and mRNA in scleral fibroblasts of the rats in FDM group were decreased(P<0.01), and the expression levels of DCL3, p21-GSK3β, and β-catenin protein and mRNA were increased(P<0.05).Compared with FDM group, the expression levels of TGF-β1 and APC protein and mRNA in the scleral fibroblasts of the rats in FDM+DDK1 group were increased(P<0.01), and the expression levels of DCL3, p21-GSK3β, and β-catenin protein and mRNA were decreased(P<0.01). Conclusion:The expression level of TGF-β1 in the scleral fibroblasts of the FDM model rats is decreased, and its level is regulated by the Wnt/β-catenin signaling pathway.

Objective:To explore the influence of tanshinone ⅡA on the expressions of molecular chaperone Cosmc, advanced oxidation protein products (AOPP) in the kidney tissue of the allergic purpura nephritis mice and the levels of extracellular signal-regulated kinase/mitogen activated protein kinase (ERK/MAPK) signaling pathway proteins,and to clarify mechanism of tanshinone Ⅱ A in the treatment of allergic purpura nephritis. Methods:A total of 38 mice were divided into control group, model group (allergic purpura nephritis model) and tanshinone Ⅱ A group (allergic purpura nephritis model + tanshinone Ⅱ A treatment), with 12 in each group; the other 2 mice were used to verify the success of modeling. The urine red blood cell counts and urine protein levels of the mice in various groups were measured, and the pathomorphology of kidney tissue of the mice in various groups were observed by periodate schiff-reaction (PAS) staining. The AOPP protein levels in the kidney tissue of the mice were determined by enzyme linked immunosorbent assay (ELISA), and the ERK and phosphorylated ERK (p-ERk) protein expression levels were determined by Western blotting method. The expression levels of ERK, p-ERK and Cosmc mRNA in kidney tissue of the mice were determined by RT-PCR. Results:In control group, there was no hyperplasia of glomerular basement membrane and no glomerular sclerosis;in model group, the proliferation of glomerular basement membrane was obvious, the inflammatory cells were infiltrated, and glomerular sclerosis appeared;in tanshinone ⅡA group, glomerular basement membrane hyperplasia and inflammatory infiltration were not obvious in the mice, there was only small amount of infiltrated inflammatory cells. Compared with control group, the urine red blood cell count,the 24 h urine protein level,the p-ERK mRNA and AOPP protein levels of the mice in model group and tanshinone Ⅱ A group were increased(P<0.05 or P<0.01); the Cosmc mRNA expression levels were decreased (P<0.01). Compared with model group, the urine red blood cell count, 24 h urine protein level, the p-ERK and AOPP protein levels in the kidney tissue of the mice in tanshinone Ⅱ A group were decreased (P<0.05 or P<0.01), and the Cosmc mRNA expression level was increased (P<0.01). In model group, the Cosmc mRNA expression level in the kidney tissue was negatively correlated with the expression levels of AOPP protein and the expression level of p-ERK protein (r=-0.573, P<0.01; r=-0.602, P<0.01), and the level of AOPP protein was positively correlated with the expression level of p-ERK protein (r=0.614, P<0.01). Conclusion:Tanshinone Ⅱ A may play a role in the treatment of allergic purpura nephritis in the mice by lowering the levels of Cosmc,increasing the AOPP in kidney tissue and activating the ERK/MAPK signaling pathways.

Objective:To investigate the effect of berberine on the angiotensin Ⅱ (AngⅡ)-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and its mechanism,and to clarify the anti-AS mechanism of berberine. Methods:The HUVECs cultured in vitro were divided into control group (untreated), AngⅡ group (treated with 1 μmoL·L^-1 Ang Ⅱ for 24 h) and low,medium,and high doses of berberine groups (after pretreated with 1 μmoL·L^-1 Ang Ⅱ for 3 h, treated with 25, 50, and 100 μmoL·L^-1 berberine for 24 h, respectively).The survival rates and the apoptotic rates of cells in each group were detected by MTT method and flow cytometry,the ROS level in each group was detected by ROS kit, and the expression levels of p-JNK, Bcl-2, Bax, and cleaved caspase-3 proteins in each group were detected by Western blotting method. Results:Compared with control group, the survival rate of cells and the Bcl-2 protein expression level in AngⅡgroup were decreased(P<0.05), while the apoptotic rate of cells, the ROS level and the expression levels of p-JNK, Bax, and cleaved caspase-3 proteins were increased(P<0.05); compared with AngⅡgroup, the survival rates of cells and the expression levels of Bax protein in low,medium, and high doses of berberine groups were significantly increased(P<0.05), while the apoptotic rates of cells, the ROS levels and the expression levels of p-JNK, Bax, and cleaved caspase-3 proteins were decreased(P<0.05) in a concentration-dependent manner. Conclusion:Berberine can inhibit the apoptosis of HUVECs induced by AngⅡ, which may be related to the inhibition of ROS/JNK signaling pathway activation.

Objective:To investigate the effect of curcumin on the implant osseointegration in the rats with osteoporosis, and to clarify its mechanism. Methods:A total of 45 rats were randomly divided into control group (screw implant), model group (osteoporosis model + screw implant) and curcumin group (osteoporosis model + screw implant + curcumin treatment), with 15 rats in each group. The micro-CT was used to examine the relative bone volume fraction (BV/TV), average bone trabecular thickness (Tb. Th), trabecular space (Tb.Sp), average bone trabecular number (Tb.N),osseointegration index(OI) of the bone tissue around the implants of the rats in various groups; methylene blue-basic fuchsin staining was used to examine the morphology of bone tissue;the bone contact rate and the bone mass in cancellous bone area were calculated. The torque tester was used to measure the implant dislocation torque. Results:Compared with control group, the BV/TV, Tb.Th, Tb.N, OI,the bone contact rates,the bone mass in cancellous bone area,and the dislocation torque of bone interface around the implants of the rats in model group and curcumin group were significantly decreased (P<0.01), while Tb.Sp in model group and curcumin group were significantly increased (P<0.01). Compared with model group, the BV/TV, Tb.Th, Tb.N, OI, the bone contact rate, the bone mass in cancellous bone area,and the dislocation torque of bone interface around the implants of the rats in curcumin group were increased (P<0.01),while Tb.Sp was significantly decreased (P<0.01). In control group, the bonding of implant and bone was close, and the bone plate and trabeculae at the bonding interface of implant and bone were thicker and denser. In model group, the bone plate at the bonding interface of implant and bone was thin and the trabecular bone was sparse. In curcumin group, the bone plate and trabeculae at bonding interface of the implant and bone of the rats were thicker and denser. Conclusion:Curcumin can promote the implant healing, improve the implant and bone bonding strength and increase the implant stability in the osteoporosis rats.

Objective:To investigate the significance of speckle tracing echocardiography in the examination of the rat models of myocardial infarction, and to elucidate the reliability and clinical application of speckle tracing echocardiography in the identification of the myocardial infarction rats. Methods:A total of 50 rats were randonly divided into sham operation group and model group,with 25 rats in each group. Ischemic preconditioning method was used to establish the myocardial infarction models. Speckle tracking echocardiography and cardiac tissue Masson staining were performed to analyze the changes of echocardiography and the morphology of myocardium tissue of the rats. Results:Compared with sham operation group,the each index in echocardiography of the rats in model group before modeling had no significant differences (P>0.05). Compared with sham operation group, the left ventricular ejection fraction (EF), circumferential strain (CS), the peak values of CS in the anterior septum, anterior wall, anterior lateral wall, systolic period, radial strain (RS) of the anterior wall of the rats in model group were decreased(P<0.05); the standardized time to peak of CS of anterior wall(TTP) was increased(P<0.05). The results of Masson staining showed that there was no change in the myocardium tissue and no infarction of the rats in control group;the infarction area of the rats in model group was (4.52±1.41)%;the infarction located in the anterior wall which was the myocardiocyte lesion in the small area. Conclusion:Speckle tracing echocardiography RS and TTP can reveal the infarction area and its surrounding areas of myocardial infarction, and quantitatively detect the myocardial infarction site in the rats. Speckle tracing echocardiography can be used for the clinical rapid diagnosis of myocardial infarction and confirmation of infarction area.

Objective:To detect the serum levels of CTRP3 and 25(OH) D in the individuals with different glucose metablism states,and to discuss the influence factors and analyze their relationships with insulin resistance(IR). Methods:Forty-four patients newly diagnosed T2DM(T2DM group),42 patients with impaired glucose regulation(IGT group),and 54 cases of normal controls(normal control group) were selected.The serum CTRP3, FPG,HbA1c,FIns,TG,TC,HDL-C, LDL-C,FIns of the subjects in various groups were determined, respectively;the BMI,HOMA-IR,HOMA-β were calculated.Pearson correlation analysis was used to analyze the correlation between two factors and multivariate stepwise regression analysis was used to analyze the influencing factors of HOMA-IR and HOMA-β. Results:Compared with normal control group, the serum levels of HDL-C, 25(OH)D and HOMA-β of the patients in IGT group were significantly decreased (P<0.05); the levels of serum FPG, HbA1c, TG, TC, FIns and BMI, HOMA-IR of the patients in T2DM group were significantly increased (P<0.05), while the levels of serum CTRP3, HDL-C, 25(OH)D and HOMA-β were significantly decreased(P<0.05). Compared with IGT group, the levels of serum FPG, HbA1c, TG, FIns, and HOMA-IR of the patients in T2DM group were significantly increased (P<0.05), and the level of serum HDL-C and HOMA-β were significantly decreased (P<0.05).The serum CTRP3 level was negatively correlated with the levels of HbA1c, FIns and HOMA-IR (r=-0.391, P<0.05; r=-0.198, P<0.05; r=-0.481, P<0.05); the 25(OH)D level was negatively correlated with the TG, FPG levels and HOMA-IR (r=-0.209, P<0.05; r=-0.406, P<0.05; r=-0.306, P<0.05), and positively correlated with HOMA-β (r=0.329, P<0.05) in the different glucose tolerance individuals. HbA1c and CTRP3 were the independent influencing factors of HOMA-IR, and 25(OH)D, FPG and HbA1c were the independent influencing factors of HOMA-β. Conclusion:The serum CTRP3 and 25(OH)D levels are negatively correlated with IR,and they have the inhibitory effects in the occurrence and development of diabetes mellitns.

Objective:To investigate the expression of NAD-dependent protein deacetylase sirtuin-6(SIRT6) protein in the colon cancer tissue and the association between its expression and the clinicopathological parameters of the patients with colon cancer, and to elucidate the effect of the SIRT6 protein in the occurrence and development of colon cancer. Methods:The colon cancer tissue and the paracancerous tissue obtained from 100 patients who did not receive radiotherapy and chemotherapy before operation were selected.The expression levels SIRT6 protein in colon cancer tissue and paracancerous tissue were detected by tissue microarray technique and immunohistochemical SP method. The relationships between the expression of SIRT6 and the clinicopathological parameters of the patents with colon cancer were analyzed with rank sum test. Kaplan-Meier survival curre,univariate analysis and multivariate survival analysis were used to investigate the relationships between the clinicopathological parameters and the prognosis in the patients with colon cancer. Results:The expression levels of SIRT6 in cytoplasm and nucleus in colon cancer tissue were significantly higher than those in paracancerous tissue (Z=-4.603,P=0.000;Z=-7.610,P=0.000).The Kaplan-Meier univariate and multivariate analysis showed that the prognosis of the patients with high expression of SIRT6 in the nucleus was better, and the expression of SIRT6 was identified as the independent prognostic factor of colon cancer(HR=2.345, P=0.003); the patients with younger age, lower T and N stages had higher survival rates (HR=0.394,P=0.004; HR=2.301,P=0.012; HR=2.423,P=0.048);there was no correlation between the high expression of SIRT6 in cytoplasm and the prognosis (HR=1.309, P=0.333) of the patients with colon cancer. Conclusion:SIRT6 protein is highly expressed in the nucleus of colon cancer and it can be used as a good prognostic indicator for colon cancer.

Objective:To analyze the differents in the upper airway morphology and hyoid position between skeletal class Ⅲ malocclusion of high-angle and normal occlusion by cone beam CT(CBCT), and to study the influence of skeletal class Ⅲ malocclusion of high-angle in the upper airway and hyoid position of the adults preliminarily. Methods:A total of 42 adults in Department of Orthodontics, Dalian Stomatology Hospital were chosen, including 21 adults with skeletal class Ⅲ malocclusion of high-angle and 21 adults with normal occlusion. MIMICS 20.0 software was used to measure the line spacing, cross-sectional area and volume of each upper airway segment and line distance of hyoid of the patients on CBCT; SPSS 20.0 software was used for statistical analysis. Results:Compared with normal occlusion group, the maximum lateral distance (LAT1) of the nasopharynx, the maximum anterior-posterior distance (AP2) of the velopharyngeal, and the volume of the velopharyngeal (VOL2) of the patients in skeletal class Ⅲ malocclusion of high-angle group were increased (P<0.05). Compared with normal occlusion group, the maximum lateral distance of the glossopharynx and laryngopharynx (LAT3 and LAT4), the cross-sectional area of the laryngopharynx (CSA4) and the volume of the laryngopharynx (VOL4) of the patients in skeletal class Ⅲ malocclusion of high-angle group were decreased (P<0.05). While no statistically significant difference was found in the position of the hyoid bone that had a tendency to shift forward and upward of the patients in skeletal class Ⅲ malocclusion of high-angle group compared with normal occlusion group (P>0.05). Conclusion:Cross-sectional area and volume of velopharyngeal have the tendency of increase, but cross-sectional area and volume of laryngopharynx have the tendency of decrease in the patients with skeletal class Ⅲ malocclusion of high-angle. The hyoid bone has a tendency to shift forward and upward in the patients with skeletal class Ⅲ malocclusion of high-angle.

Objective:To investigate the value of risk ovarian malignancy algorithm (ROMA) in evaluation of the risk of epithelial ovarian cancer and its association with the clinical pathological stages, and to provide a basis for the clinical diagnosis and treatment and prognosis analysis of epithelial ovarian cancer. Methods:The clinical materials of 135 patients with ovarian tumor confirmed by paraffin section pathology after operation were retrospectively analyzed. The patients were divided into benign ovarian tumor group(n=66), epithelial ovarian cancer group(n=58) and non-epithelial ovarian cancer group(n=11).According to the cutoff values of HE4,CA125,and ROMA of the patients in various groups,the positive rates of HE4,CA125,and ROMA of the patients in various groups were calculated.Based on the serum levels of human epididymis protein 4(HE4) and carbohydrate antigen 125(CA125) detected before operation of the patients in various groups, the relationships between the positive rates of serum HE4, CA125 and ROMA and the clinical stages of the patients with ovarian cancer;their diagnosis efficacies in the patients with ovarian cancer were evaluated. Results:The serum CA125, HE4 levels and ROMA value of the patients in epithelial ovarian cancer group were significantly higher than those in benign ovarian tumor group and non-epithelial ovarian cancer group (P<0.05). The positive rates of HE4, CA125, and ROMA of the patients in epithelial ovarian cancer group were higher than those in benign ovarian tumor group (P<0.01), and showed an increasing trend with the increase of clinical stages.Compared with the positive rates of HE4 and CA125, the ROMA had the lowest positive rate in the patients with ovarian benign tumor, and the positive rates of ROMA in all stages of epithelial ovarian cancer were higher than those of CA125.The sensitivity and the negative predictive value of ROMA in postmenopausal patients with ovarian cancer group were higher than those in premenopausal patients with ovarian cancer group (P<0.05), while the specificity and the positive predictive value were not significantly different between premenopausal and postmenopausal patients with ovarian cancer groups (P>0.05). Among the three indexes, ROMA had the highest diagnostic efficacy index and CA125 had the lowest. Conclusion:The diagnostic value of ROMA for evaluating the risk of epithelial ovarian cancer was higher than those of CA125 and HE4, which has a better diagnostic value for the postmenopausal patients and can improve the early diagnosis efficiency of ovarian cancer.

Objective:To investigate the effect of citric acid on the salivary gland function in the patients with differentiated thyroid cancer who received ¹³¹I treatment for the first time after operation, and to clarify the protective effect of citric acid on the salivary gland function in the patients with thyroid cancer who received¹³¹Itreatment. Methods:With the informed consent of the patients, 68 patients with thyroid papillary carcinoma who were going to receive ¹³¹I treatment were randomly selected and divided into control group and citric acid group (n=34). There was no special preparation for the patients in control group, and the patients in citric acid group kept citric acid in the mouth for 1 min and spit it out every day (0.2 g/time) at 1 week before and 3 weeks after ¹³¹I treatment. The patients in two groups received ^99mTcO^4- salivary gland imaging twice within 24 h before ¹³¹I treatment and 3 months after ¹³¹I treatment. The uptake index in the fifteenth minute (15 min UI) and the secretion ratio (SR) were calculated and the salivary gland function was evaluated. Results:Compared with before ¹³¹I treatment, the 15 min UI of the right parotid gland and bilateral submandibular glands of the patients in control group after treatment had no significant differences(P>0.05); the 15 min UI of left parotid gland after treatment was slightly decreased(P<0.05);compared with before ¹³¹I treatment, the 15 min UI of the bilateral parotid glands and bilateral submandibular glands of the patients in citric acid group had no significant differences(P>0.05). Compared with control group, the 15 min UI of bilateral parotid glands and bilateral submandibular glands of the patients in citric acid group before and after treatment had no significant differences(P>0.05). Compared with before ¹³¹I treatment, the SR of bilateral parotid glands of the patients in control group after treatment were decreased (P<0.05), and there were no differences in the SR of bilateral submaxillary glands after treatment(P>0.05). Compared with before ¹³¹I treatment, the SR of bilateral parotid glands and bilateral submandibular glands of the patients in citric group after treatment had no significant differences(P>0.05). The SR of bilateral parotid glands of the patients in citric acid group after treatment were higher than those in control group (P<0.05), but there were no significant differences in the SR of bilateral submandibular glands of the patients between two groups after treatment (P>0.05). Conclusion:First ¹³¹I treatment may cause the damage of the secretion function of the salivary glands in the DTC patients after operation. Short-term administration of citric acid in the mouth can protect the salivary glands and alleviate the radioactive injury of the salivary glands.

Objective:To analyze the postoperative satisfaction and repaireffect of prosthesis of the patients who received digital smile design(DSD) combined with porcelain veneers in the anterior teeth,and to illustrate the their application in anterior teeth aesthetic restoration. Methods:A total of 91 anterior teeth of 32 patients whose clinical diagnosis conformed to the indications of porcelain veneers and aesthetic restoration because of tetracycline pigmentation teeth, dental fluorosis, color stained teeth, enamel defects and anterior teeth space were selected. DSD software was used to design the preoperative aesthetic analysis of the teeth morphology and to show the virtual effect of restoration. The preparation of tooth under the guidance of the design results was finished. The IPS e-max porcelain veneers were prepared after the conventional impression preparation. At last, the permanent restorations were manufactured and bonded to teeth. After the adhesive program was completed, the satisfaction of the patients in the shape of restorations, the degree of coordination with the adjacent teeth, color, pronunciation, smile effect and doctor-patient communication effect were evaluated by the questionnaires. The patients were required to regularly return after one month, three months, six months and twelve months,and observe the clinical effects of restoration was observed referring to the modified American Public Health Service (USPHS) standard. Results:The patients' satisfaction rates of each evaluation content were all over 90%. Among them, the satisfaction rates of the smile effect and the doctor patient communication were as high as 100%. The clinical performances of the rehabilitations in each period were as follows:one month after treatment,there were four teeth occurred mild gingivitis and three teeth appeared the postoperative sensitivity symptoms; three months after treatment, there were one tooth showed a minor fracture in the restoration without affecting the appearance and function and three teeth had mildly congestive gums reaction, at the same time one tooth showed slight discoloration of the restoration; six months after treatment, one tooth showed ceramics fracture of restoration and four teeth developed mild gingivitis; twelve months after treatment, two teeth showed ceramics fracture, the edge of three restorations had slightly crack and one tooth appeared gingivitis. The periodontal conditions of the patients with gingival inflammation after treatment were improved significantly after receiving the correct oral hygiene guidance. The above defects didn't affect the aesthetics effect and the functions of restorations, and the restorations could be used normally after intraoral adjustments. Both the doctors and the patients were satisfied with the repair effects of the restorations. Conclusion:The combination of DSD and porcelain veneer in repairing anterior aesthetic teeth is good treatment plan that can achieve high patient satisfaction and ideal clinical effect.

Objective:To compare the scores of periocular wrinkle and blepharochalasis of the patients before and after the treatment of ultra-pulsed fractional CO₂ laser with low energy,and to illuminate the efficacy of ultra-pulsed fractional CO₂ laser with lower energy in eyelid rejuvenation including periocular wrinkle and blepharochalasis. Methods:Thirty patients with periocular wrinkles and blepharochalasis who met the inclusion criteria were selected. The patients were treated with ultra-pulsed fractional CO₂ laser; the laser parameters were 10 mm spot size, pulse energy 5 mJ and repetition rate 600 Hz and the wrinkle walking areas were treated with 2 mm spot size and the same energy again. Two senior dermatologists scored according to the eyelid laxity scale before treatment and 7 d, 1 month, 3 months and 6 months after treatment, and statistical analysis was performed. Results:Compared with before treatment,there was no significant difference in the scores of eyelid laxity scale 7 d after treatment(P>0.05); the score of eyelid laxity scale was significantly decreased 1 month after treatment (P<0.05);the scores of eyelid laxity scale were significantly decreased 3-6 months after treatment (P<0.01),and the curative effect was more obvious.In this group of 30 patients, there were usually transient erythema, swelling and mild burning pain after treatment. But no serious complications occurred. Conclusion:Ultra-pulsed fractional CO₂ laser with lower energy(5 mJ) can improve the periocular wrinkles and blepharochalasis effectively with high security and without severe side effects.

Objective:To investigate the surgical technique of autologous tendon transplantation for reconstruction of anterior cruciate ligament (ACL) injury,and to analyze its curative effect. Methods:The materials of 30 patients with ACL injury who underwent autologous tendon transplantation and reconstruction were retrospectively analyzed; the average follow-up period of the patients was 24.1 months. The surgery characteristrics and clinical curative effect were explored. Results:The stability of knee joint was examined by drawer test,Lachman test,and internal and external stress tests. The healing of reconstructed ligament tunnel was observed by X-ray. The morphology of ACL and ligament reconstruction before and after operation were observed by magnetic resonance imaging (MRI). The surgical effect was evaluated according to the International Knee Documentation Committee (IKDC) scoring system. There were no severe complications such as traumatic arthritis, infection, or thrombosis in the patients. The results of anterior drawer test, Lachman test, and internal and external stress tests were changed from positive to negative, suggesting that the knee joint was stable after operation. The postoperative IKDC score of the patients was significantly higher than before operation(P<0.05). The X-ray results showed that the blurred or disappeared bone tunnel basically achieved tendon-bone healing, while the MRI showed that the reconstructed ligament signal was uniform and non-tortuous, indicating that the ligament was strengthened. Conclusion:Accurate reconstruction with autologous ligament transplantation in the treatment of ACL injury is safe and feasible with fewer complications, which is helpful to restore the stability of knee joint quickly.

Objective:To observe the clinical manifestations of cervical esophageal webbed stenosis in thepatients with Plummer-Vinson syndrome(PVS) and analyze its relationship with hypopharyngeal carcinoma, and to raise the clinicians' awareness of the disease. Methods:The diagnosis and treatment process of one patient with PVS complicated with hypopharyngeal carcinoma was summarized, and its clinical characteristics were analyzed in combination with relevant literatures. Results:A 39-year-old patient with PVS as a complaint by "dysphagia" was diagnosed as hypopharyngeal carcinoma by electronic laryngoscope and pathological biopsy. A lot of intraepithelial papillary capillary loops (IPCL) were seen on narrow band imaging(NBI) of the mucosa extending downward from the posterior and lateral walls of the hypopharynx to the upper part of the esophageal web. The hypopharyngeal mucosa with dense IPCL was found to have moderate to severe dysplasia. After 2 courses of induction chemotherapy for hypopharyngeal carcinoma, the symptoms disappeared and the tumor volume was significantly decreased. Conclusion:The occurrence of hypopharyngeal carcinoma is related to the mucosal changes of PVS. NBI technique is helpful for early diagnosis of hypopharyngeal and esophageal mucosal degeneration in the PVS patients.

Objective:To summarize the relationship between anti-Ro-52 antibody and anti-synthase syndrome(ASS)by analyzing the diagnosis and treatment of ASS patients with positive anti-Ro-52 antibody,and to clarify its significance. Methods:The clinical materials of 2 ASS patients with positive anti-Ro52 antibody were collected,and the clinical characteristics were analyzed;combined with literature review,the relationship between anti-Ro-52 antibody and ASS was explored. Results:Two young female patients were admitted to hospital because of the obvious symptoms of both hand joints. On physical examination, arthroncus of both hand joints was observed, both hands showed "mechanic hand", muscle strength and muscle tension of limbs were normal. The activity of creatine kinase was increased. Myositis antibody spectrum, lung CT, electromyography and muscle biopsy were performed and the patients were given the related treatments after confirmed diagnosis. The anti-Jo-1 and anti-Ro-52 antibodies were strongly positive in the myositis antibody spectrum of the two patients; the lung CT results showed interstitial pheumonia;the electromyogram results indicated myogenic damage; the results of muscle biopsy was consistent with the changes of idiopathic inflammatory myopathy. Both patients were diagnosed as ASS. After treatment of glucocorticoids and immunosuppressive agents, the symptoms were improved and the creatine kinase activity was decreased. No disease activity and secondary tumor signs were found in the follow-up. Conclusion:As a myositis-associated antibody,anti-Ro-52 antibody can lead to a series of atypical clinical manifestations in the ASS patients and may be associated with the poor prognosis of the ASS patients. Follow-up should be paid attention to.

Objective:To analyze the clinical features of ovarian fibrothecoma complicated with ascites and high level of CA125, and to explore the causes of elevated CA125 level and its significance in identifying benign and malignant ovarian tumors. Methods:The clinical data of one patient with varian fibrothecoma complicated with ascites and high level of serum CA125;combined with relevant literature review, the clinical characteristics, diagnosis and treatment process, and the significance of CA125 in the differential diagnosis of benign and malignant ovarian tumors were analyzed. Results:The patient was admitted to hospital due to vaginal bleeding for 4 months and pelvic mass for 1 month. The diagnosis result was pelvic and abdominal mass arising from ovarian according to the preoperative color Doppler ultrasound, enhanced CT, serum CA125 level and physical examination,and was highly suspected as ovarian malignant tumor. Surgery treatment was performed; based on the intraoperative rapid pathological results and postoperative slow pathology, the patient was diagnosed as ovarian fibrothecoma; the operation was successful. The patient recovered well and discharged from hospital.The serum CA125 level was decreased to the normal range 1 month after operation. Conclusion:Identification and quantitative analysis of the glycosylation modification and specific glycan structure of CA125 from different sources can significantly improve the diagnostic specificity of CA125 for benign and malignant ovarian tumors.

Objective:To explore the related factors of depression and anxiety among the retired employees, and to examine the mediating role of social participation quality in the elderly between living alone and depression and anxiety. Methods:The retired employees who met the admission criteria in the First Hospital of Jilin University were selected as the subjects. The basic situation questionnaire was used in this study, and the social participation quality, depression and anxiety were assessed by social aspect of the Quality of Life Scale, Patient Health Questionnaire Depression Table and Anxiety Screening Questionnaire, respectively. A total of 642 valid questionnaires were obtained. Linear regression analysis was used to analyze the influencing factors, and the mediating effect test was conducted by stepwise regression analysis. Results:The influencing factors of depression in the retired employees were physical exercise (β=-0.948, P<0.01), entertainment (β=-1.423, P<0.01), whether to suffer from chronic diseases (β=2.579, P<0.01), family income (β=-0.557, P<0.05), whether to insist on work (β=-1.673, P<0.05), and whether to live alone (β=2.097, P<0.01).The influencing factors of anxiety were physical exercise (β=-0.711, P<0.05), entertainment (β=-1.380, P<0.01), whether to suffer from chronic diseases (β=1.204, P<0.05), and whether to live alone (β=1.639, P<0.05).In the elderly, living alone status and social participation quality were input in the linear regression as the independent variables,and the results showed that the effects of living alone status on depression and anxiety had no the significant differences (β=0.837, P=0.218; β=0.837, P=0.218), and social participation quality played a full mediating role between living alone status and depression and anxiety. Conclusion:Physical exercise, entertainment, whether to suffer from chronic diseases and whether to live alone are the influencing factors of depression and anxiety in the retired employees; social participation quality plays a full mediating effect between living alone status and depression and anxiety.

Objective:To investigate the diet of lactating mothers,breast milk components and the growth of infants in Changchun City, Jilin Province and explore the relationships between the nutritional status of breast milk and the growth of infants, and to provide the theoretical basis for the development of the infant health. Methods:A total of 138 healthy nursing mothers were selected. Within 40-45 d after the delivery, face-to-face investigations were conducted to obtain the basic information of the subjects and the growth and development information of the infants. The 24-hour dietary review method was adopted to investigate the dietary status of the lactating mothers, the standard food pictures were provided to estimate the dietary variety and the quantities of the subjects. Golden key maternal nutrition expert system software was applied to analyze the energy and nutrients of dietary intake, and they were compared with Dietary Guidelines of Pregnant, Lactating Women and 0-6 Years Old Children in China (2007) and Chinese Dietary Reference Intake of Nutrients (2013);the diet intake level of the lactating mothers was evaluated. The breast milk was collected and the infant growth and development indicators were measured. The compositions of breast milk were analyzed by milk composition analyzer. The relationships between the lactating mother's diet, the breast milk composition and the infant growth and development were analyzed. Results:The average energy intake of the lactating mothers was (1 698.7±406.23) kcal·d^-1, and the energy proportions of proteins, lipids, and carbohydrates were 14.06%, 33.83%, and 52.09%, respectively. The average energy content of breast milk was (61.18±13.04) kcal·100g^-1, the average protein content was (0.94±0.19) g·100g^-1, the average lipid content was (3.55±1.27) g·100g^-1, and the average lactose content was (6.38±0.82) g·100g^-1. The dietary protein intake was associated with the protein content of breast milk (r=0.203,P=0.017).When the influence of feeding patterns, energy of breast milk, infant gender, and age were controlled, the partial correlation analysis result showed that the breast milk composition had no correlations with the increases in body lengths and weights of the infants (P>0.05). Conclusion:There are insufficience and imbalance in the general dietary intakes in the lactating mothers. The dietary protein intake of lactating mothers may influence the protein content of breast milk. The lipid and carbohydrate contents in milk have no significant correlations with the dietary nutrients of the lactating mothers. The breast milk compositions of lactating mothers have no effects on the growth and development of infants.

Objective:To coat polydopamine (PDA) on the surface of biphasic calcium phosphate (BCP) scaffold prepared by 3D printing and construct a novel bone tissue engineering scaffold with high osteogenic activity,and to conduct a preliminary evaluation on its application potential. Methods:BCP bioceramic scaffolds were prepared by 3D printing technology, and then they were immersed in dopamine solution for a certain period to form a nanoscale PDA film structure on the surface.The PDA-free scaffolds were named 3DBCP group, and the PDA-coated scaffolds were named PDA-3DBCP group. Scanning electron microscope (SEM) was used to observe the surface topography of the scaffolds in each group; the hydrophilicity of the material was characterized by measuring the water contact angle of the scaffold surface; the scaffold porosity was determined by the specific gravity method, and the mechanical strength was tested by the universal testing machine; CCK-8 assay was adopted to detect the cell proliferation activity on the scaffold. Results:Compared with 3DBCP group, the surface water contact angle of the the scaffolds in PDA-3DBCP group was significantly reduced (t=5.06, P<0.05);there were no significant differences in the porosity and the mechanical strength between two groups(t=0.103, P>0.05; t=0.002, P>0.05).The cells were inoculated into the scaffolds and cultured for 1, 3 and 5 d;the CCK-8 assay results showed that the proliferation activities of the cells in two groups were gradually increased with the prolongation of the culture time. Furthermore, compared with 3DBCP group, the cell proliferation activity of the cells in PDA-3DBCP group on the 5th day was significantly increased (t=39.3, P<0.05). Conclusion:The PDA-coated 3D printed BCP porous scaffold has the advantages in characterization,it can improve the cell proliferation ability, which shows the potential as bone tissue engineering scaffolds.