Objective: To observe the effect of CoCl₂ on the cisplatin sensitivity of human ovarian cancer SKOV3 cells, and to clarify the possible mechanism.Methods: The SKOV3 cells were cultured in vitro and randomly divided into control group, CoCl₂ group, cisplatin (DDP) group and CoCl₂ combined with DDP(combination) group. The cells in CoCl₂ group were cultured in normal cell medium for 20 h after cultured in 200 μmol·L^-1 CoCl₂ for 4 h, the cells in DDP group were cultured in normal cell medium containing 10 mg·L^-1 DDP for 24 h, and the cells in combination group were cultured in 10 mg·L^-1 DDP for 20 h after cultured in 200 μmol·L^-1 CoCl₂ for 4 h. The survival rates of SKOV3 cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1 α (HIF-1α) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method. Rhod 2-AM fluorescence probe was used to observe the levels of Ca²⁺ in mitochondria in the cells in various groups. Western blotting method was used to observe the expression levels of cytochrome C (cyto C), cysteinyl aspartase 3 (caspase 3) and cleaved cysteinyl aspartase 3 (cleaved caspase 3). Muse^® apoptosis assay kit was used to detect the apoptotic rates of cells in various groups.Results: Compared with control group, the survival rate of the cells in CoCl₂ group had no significant change (P>0.05), and the survival rates of the cells in DDP and combination groups were decreased (P<0.05); the survival rate in combination group was higher than that in DDP group (P<0.05). Compared with control group, the positive expression intensities of HIF-1α in CoCl₂ and combination groups were increased (P<0.05). Compared with control group, the positive expressions of iNOS in DDP and combination groups were increased (P<0.05). The Ca²⁺ levels in the cells in DDP group and combination groups were higher than that in control group (P<0.05) and the Ca²⁺ level in DDP group was higher than that in combination group (P<0.05). Compared with control group, the expression levels of cyto C,caspase 3 and cleaved caspase 3 proteins in the SKOV3 cells in CoCl₂ group had no significant changes (P>0.05), and the expression levels of cyto C,caspase 3 and cleaved caspase 3 in DDP group were increased significantly(P<0.05); compared with DDP group, they were lower than those in combination group (P<0.05). Compared with control group, the apoptotic rate of SKOV3 cells in DDP group was increased significantly(P<0.05); the apoptotic rate of SKOV3 cells in combination group was lower than that in DDP group (P<0.05).Conclusion: CoCl₂ can redece the mitochondrial apoptosis of human ovarian cancer SKOV3 cells by inhibiting the DDP-induced enhancement of iNOS expression and decrease the sensitivity of SKOV3 cells to cisplatin.

Objective: To investigate the regulatory effect of lumbrukinase (LBK) on the gastric cancer SGC7901 cells, and to clarify its mechanism.Methods: The SGC7901 cells in the logarithmic growth phase were selected and divided into control group and 2,4,8 U·mL^-1 LBK groups. MTT assay was used to detect the inhibitory rates of proliferation of SGC7901 cells in various groups in vitro at different time (24, 48 and 72 h). Cell scratch assay was used to detect the migration abilities of the SGC7901 cells in vitro in various groups.Flow cytometry was used to determine the apoptotic rates of SGC7901 cells and the pencentages of cells at different cell cycles in various groups. The expression levels of Bcl-2, Bax, and caspase-3 in the SGC7901 cells in various groups were detected by Western blotting method.Results: The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901 cells in different doses of LBK groups after treated for 24,48 and 72 h were increased(P<0.01). The cell scratch assay results showed that compared with control group, the migration distances of SGC7901 cells in 4 and 8 U·mL^-1 LBK groups were increased significantly(P<0.01).The flow cytometry results showed that compared with control group,the apoptotic rates of SGC7901 cells in 4 and 8 U·mL^-1 LBK groups were increased significantly(P<0.01);the percentages of cells in G₁ and S phases were decreased(P<0.01);the percentages of cells in G₂ phase were increased(P<0.01).The results of Western blotting method showed that compared with control group, the Bcl-2 protein expression level in the SGC7901 cells in 8 U·mL^-1 LBK group was decreased (P<0.05); the Bax and caspase-3 protein expression levels were increased (P<0.05).Conclusion: LBK can inhibit the proliferation and migration abilities of SGC7901 cells in vitro and induce the apoptosis; its mechanism is achieved through the regulation of expression levels of Bcl-2, Bax, and caspase-3 proteins.

Objective: To reveal the differentially expressed genes and long non-coding RNAs (lncRNAs) by sequencing the transcriptome of bovine alveolar macrophages (BAM) infected with Bacillus Calmette-Guérin (BCG) and analyzing their bioinformations, and to provide theoretical foundation for the research on immune regulation mechanism of anti-infection of Mycobacterium tuberculosis of macrophages.Methods: The BAM were collected by pulmonary lavage and centrifugation and cultured and divided into infected group and uninfected group.After infection for 12 h in infected group,the expression profiles of mRNA and lncRNA in infected group and uninfected group were detected by RNA-Seq, and bioinformatics analysis was carried out.Results: compared with uninfected group, there were 119 differentially expressed lncRNAs and 1111 differentially expressed mRNA in infected group (P<0.05). Gene Ontology functional enrichment analysis showed that the most significant enrichment was immune response (GO:0006955, P<0.05), including 125 genes, in which 63 were up-regulated and 62 were down-regulated, and the expressions of proinflammatory factors interleukin-1 (IL-6), interleukin-7 (IL-7), and interleukin-23A (IL-23A) were up-regulated. Cis target gene prediction and KEGG pathway analysis showed that the differentially expressed lncRNAs were involved in the regulation of transforming growth factor-β (TGF-β) signaling pathway and ATP-binding cassette transporter (ABC transporter) signaling pathway (P<0.05).Conclusion: The host cells are stimulated to produce a strong immune response after the BAM are infected by BCG and results in the changes of lncRNA and mRNA expression profiles.

Objective:To explore the effects of the biomimetic network membrane prepared by chitosan/gelatin/pectin on the proliferation and mineralization of mesenchymal stem cells (MSCs), and to evaluate its feasibility of constructing tissue engineering bone.Methods: Chitosan, gelatin and pectin were made into a new biomimetic network membrane in a certain ratio by biomimetics.The experiment was divided into control group(MSCs+conventional medium),material group(MSCs+network membrane+conventional medium)and material+OS group(MSCs+ network membrane+OS medium).The cell morphology was observed by inverted phase contrast microscope; the growth and secretion of extracellular matrix of the MSCs were observed under scanning electron microscope(SEM).The proliferation of cells was determined by MTT assay(The MSCs were divided into negative control group and material group,and they were cultivated with blank medium and medium including materials).The expression of calcium in MSCs was detected by Alizarin Red staining.Real-time polymerase chain reaction (RT-PCR) was used to determine the expression levels of osteocalcin (OC) mRNA and osteopontin (OPN) mRNA in the MSCs.Results: The network membrane was semitransparent thin film.The MSCs were short shuttle and clustered under inverted phase contrast microscope.After cultured for 7 d,the MSCs were shuttle; after cultured for 14 d,the number of MSCs was increased,with pseudo feet on the membrane;after cultured for 21 d,the MSCs clustered with a lot of neo-formed extracellular matrix.The MTT results showed that there was no significant difference in the proliferation level of MSCs between material group and negative control group (P>0.05).The Alizarin Red staining results showed that the MSCs in the network membrane were dyed orange red.The RT-PCR results showed that the expression levels of OC mRNA in the MSCs in material group and material+OS group were lower on the 7th and 14th days, but on the 21th day,the expression levels were significantly increased and reached the peak;the expression level of OC mRNA in the MSCs in material group was significantly increased on the 7th day, and the expression level reached the peak on the 14th day, then fell slightly on the 21th day;compared with control group,the expression levels of OC mRNA and OPN mRNA in the cells in material group and material+OS group at different time points were significantly increased(P<0.01),but there were no significant differences between material group and material+OS group(P>0.05).Conclusion: Chitosan/gelatin/pectin biomimetic network membrane has good biocompatibility,and MSCs can grow and proliferate well on the membrane.The membrane can induce the MSCs to express mineralization-related genes and proteins without inducers.

Objective: To detect the mRNA expression levels of inflammatory-related factors NLRP3, caspase-1, IL-1β, and IL-18 in the murine macrophages infected by periodontitis patient's own tissue nucleic acid, and to discuss the effects of periodontitis patient's own tissue nucleic acid on the inflammation-related factors in the macrophages.Methods: The inflammatory periodontal tissue samples were collected during periodontal flap surgery of the chronic periodontitis patients, and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery. Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA. The cultured mouse macrophages RAW264.7 were divided into control group and experiment group, then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA (the cDNA at a concentration of 1 mg·L^-1)were added into the RAW264.7 cells,respectively. Real-time PCR was used to detect the mRNA expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 in the macrophages in various groups at 4, 6 and 8 h after incubation.Results: The microscope observation showed that the mouse macrophages RAW264.7 grew well with round and polygon shapes, clear cytoplasm, and full cell body. Compared with control group, the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 mRNA in the RAW264.7 cells in experiment group at 4, 6, and 8 h were increased significantly(P<0.05 or P<0.01), and the expression levels of NLRP3,Caspase-1,IL-1β, and IL-18 mRNA in RAW264.7 cells at 6 h in experiment group were the highest.Conclusion: The periodontitis patient's own tissue nucleic acids can promote the mRNA expressions of inflammation-related factors in the RAW264.7 cells, suggesting that the periodontitis patient's own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264.7 cells.

Objective: To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1 cells,and to clarify its mechanism.Methods: Three kinds of human esophageal cancer cells, TE-1, ECA109 and KYSE150, were selected. The relative expression levels of MRPL35 mRNA in three kinds of cells by real-time quantitative PCR. The esophageal cancer TE-1 cells were divided into shMRPL35 group and shCtrl group,and the cells were infected with si-RNA lentivirus and si-RNA lentivirus; the esophageal cancer cell line stably silenting the MRPL35 gene was established. Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35 gene silencing.The cell growth curves in various groups were detected by CCK-8 method, and the apoptotic rates were detected by flow cytometry after Annexin Ⅴ-PE/7AAD double staining.Results: Three kinds of esophageal cancer cells expressed MRPL35 gene, and the expression levels were not statistically significant between them(P>0.05). The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35 in the TE-1 cells in shMRPL35 group were significantly lower than those in shCtrl group(P<0.05). Compared with shCtrl group, the cell growth speed in shMRPL35 group was decreased (P<0.05), and the apoptotic rate was significantly increased (P<0.01).Conclusion: Silencing MRPL35 gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.

Objective: To investigate the induction of tanshinone ⅡA (TanⅡA) on the differentiation of human placenta-derived mesenchymal stem cells (hPDMSCs) into cardiomyocytes, and to provide an experimental basis for TanⅡA as a cardiomyocyte differentiation inducer.Methods: The hPDMSCs were treated with different concentrations of TanⅡA(0.1,0.2,0.4,0.6,0.8,1.0,2.0,4.0,6.0,8.0,and 10.0 mg·L^-1), and the nontoxic dose of TanⅡA(0.1 mg·L^-1) was screened by MTT assay for experiment.The hPDMSCs were divided into control group, 5-aza induction (10μmol·L^-1) group, and TanⅡA induction (0.1 mg·L^-1) group.After culture for 20 d,the expressions of α-sarcomeric actin(α-SCA) in the cells in various groups were detected with immunohistochemistry;the positive expression rates of cardiac troponin I(cTnI) in the cells in various groups were detected with immunofluorescence,and the differentation rates of cardiomyocytes were calculated.The expression levels of GATA-binding protein 4(GATA4), atrial natriuretic factor(ANF), cTnI, glycogen synthase kinase-3β(GSK-3β)and β-catenin in the cells were detected with Western blotting method.Results: The biological characteristics of hPDMSCs accorded with the mesenchymal stem cells. The MTT results showed that when the concentration of TanⅡ A was more than 0.1 mg·L^-1, the cell survival rates were decreased with the increase of concentration; the cells in control group showed a rapid growth trend before 12 d, and the proliferation activities of the cells began to decrease on the 12th day.Compared with control group,the cell activities in 5-aza induction group and TanⅡ A induction group were significantly decreased(P<0.05).The immunohistochemistry staining results showed that the cells in control group didn't express α-SCA,and the cells in 5-aza induction group and TanⅡ A induction group expressed α-SCA, especially in TanⅡ A induction group.Compared with control group, the expression levels of GATA4(t_5-aza=2.937,P_5-aza<0.05;t_TanⅡA=4.769,P_TanⅡA<0.05), ANF(t_5-aza=3.728,P_5-aza<0.05;t_TanⅡA=5.912,P_TanⅡA<0.05), cTnI(t_5-aza=3.623,P_5-aza<0.05;t_TanⅡA=7.153,P_TanⅡA<0.05)and GSK-3β(t_5-aza=2.995,P_5-aza<0.05;t_TanⅡA=5.420,P_TanⅡA<0.05)proteins in the cells in 5-aza induction group and TanⅡA induction group were significantly increased, and the expression levels of β-catenin(t_5-aza=2.985,P_5-aza<0.05;t_TanⅡA=6.951,P_TanⅡA<0.05)protein were significantly decreased;compared with 5-aza induction group,the expression levels of GATA4,ANF,and GSK-3β proteins in TanⅡA induction group were increased(P<0.05).Conclusion: TanⅡA can induce the differentiation of hPDMSCs into cardiomyocytes,which has better effect than 5-aza,and its mechanism may be related to inhibiting the Wnt/β-catenin signaling pathway.

Objective: To observe the inhibitory effect of metformin on the inflammatory response of Kupffer cells(KCs) of liver in the diabetic mice, and to explore its improvement on the phagocytic function of KCs, and to elucidate the protective mechanism of metformin on the KCs in the diabetic mice.Methods: The C57BLKS/J db/db mice were divided into diabetes group and diabetes + metformin group, and the C57BLKS/J db/m mice were divided into non-diabetes group and non-diabetes + metformin group, 8 mice in each group. The KCs were cultured in vitro and the changes of ultrastrustures of KCs were observed by transmission electron microscope. The levels of interleukin 6(IL-6), tumor necrosis factor α(TNF-α)and γ-interferon(INF-γ)in the KCs were detected by ELISA. The expression levels of intercellular adhesion molecule 1(ICAM-1) protein in the KCs was detected by Western blotting method. Transwell chamber assay was used to detect the neutrophil chemotaxis ability of the KCs, and the phagocytic ability of KCs was observed under inverted microscope.Results: The number of mitochondria and rough endoplasmic reticulum (RER) in the KCs of the diabetic mice was reduced,the RER expanded,the mitochondria swelled, and the lipid droplets were increased. Compared with non-diabetes group, the levels of IL-6, TNF-α, INF-γ and the expression level of ICAM-1 in the KCs of the mice in diabetes group were significantly increased (P<0.05); the neutrophil chemotaxis ability of the KCs was enhanced (P<0.05), and the phagocytic ability was decreased (P<0.05). Compared with diabetes group, the levels of IL-6, TNF-α, INF-γ and the expression level of ICAM-1 in the KCs in diabetes+metformin group were significantly decreased(P<0.05); the neutrophil chemotaxis ability was decreased (P<0.05), and the phagocytic ability was enhanced (P<0.05).Conclusion: Metformin can inhibit the inflammatory response of KCs in the diabetic mice and improve its phagocytic function and protect the KCs.

Objective: To observe the effect of sciatic nerve pulsed radiofrequency(PRF) on the activation levels of microglia cells and astrocytes in spinal dorsal horn in the chronic constriction injury(CCI) rat models, and to explore the relationships between the analgesic mechanism of sciatic nerve PRF and the activation levels of microglia cells and astrocytes in the spinal dorsal horn.Methods: Forty male SD rats were randomly divided into CCI sham-operation+RPF sham group(SS group), CCI sham-operation+RPF group(SP group), CCI+RPF sham group(CS group), CCI+RPF group(CP group)(n=10). PRF was applied on the sciatic nerve on the 4th day after CCI operation for 120 s, with a maximum temperature of 42℃.The mechanical withdrawal thresholds (MWT) and thermal paw withdrawal latencies (TWL) were measured to evaluate mechanical hyperalgesia and thermal hyperalgesia 1 d before operation (D₀) and 1,3,5,7 d after operation (D₁, D₃, D₅, D₇). Western blotting method was used to detect the protein expression levels of ionized calcium binding adapter molecule 1(Iba-1) and glial fibrillary acidic protein (GFAP) in the ipsilateral spinal dorsal horn of L3-5 after pain behavioral test(D₇).Results: Compared with SS group, the rats in CS group after CCI had the significant behavioral changes, such as hallux valgus, lameness, toe bending, and foot raising during walking; MWT and TWL were decreased significantly(P<0.01); the expression levels of Iba-1 and GFAP proteins in the ipsilateral spinal dorsal horn were increased significantly(P<0.05). Compared with CS group, the behavioral changes of the rats in CP group (D₅, D₇) such as hallux valgus, lameness, toe bending, and foot raising during walking were alleviated significantly; MWT and TWL were increased significantly(P<0.01); the Iba-1 protein expression level in spinal dorsal horn was down-regulated significantly (P<0.05) and the GFAP protein expression level did not change significantly (P>0.05).Conclusion: PRF on sciatic nerve can relieve the neuropathic pain (NP) of the CCI rat models; PRF on sciatic nerve can inhibit activation of the microglia cells in the spinal dorsal horn. The analgesic mechanism of PRF on sciatic nerve may be associated with the inhibition of the activation of microglia cells in the spinal dorsal horn.

Objective: To investigate the influence of endoplasmic reticulum stress (ERS) in the degeneration of cochlear hair cells in the type 2 diabetic mice, and to clarify its mechanism.Methods: Twenty clean Kun Ming male mice aged one month were selected and randomly divided into control group and model group(n=10).The mice in model group were injected with STZ (40 mg·kg^-1) to establish the type 2 diabetic models. The fasting blood glucose levels of the mice were measured through collecting the vena caudalis blood of the mice. Auditory brain stem response(ABR) was used to detect the ABR threshold of the mice. Otoacoustic emission (OAE) test was used to detect the OAE threshold of mice. The defect rate of mouse cochlear outer hair cells was calculated by the mouse cochlear spreading technique. The expression levels of GRP78, caspase-12, p-ERK and Nrf2 proteins were detected by Western blotting method.Results: Compared with control group, the fasting blood glucose levels of the mice in model group at the 7th and the 14th days had no significant differences(P>0.05), but the levels were increased significantly at the 21th, 28th and 35th days and the level reached the highest value at the 35th day. The ABR thresholds of the mice in model group at 8,12,and 24 kHZ were increased significantly compared with control group(P<0.05). Under the stimulation of low frequency, there was no significant change in the OAE threshold of the mice in model grouop compared with control group. The OAE thresholds of the mice in model group were increased significantly under the medium frequency and high frequency stimulation compared with control group(P<0.05).The defects of the cochlear hair cells were mainly concentrated on the bottom of gyrus of the mice,and the defects in middle temporal gyrus and parietal gyrus were less.Compared with control group,the defect rate in the bottom of gyrus of the mice in model group was increased significantly(P<0.05);the defect rates in the middle temporal gyrus and parietal gyrus were increased,but there was no significant difference(P>0.05). The expression levels of p-ERK and Nrf2 in the cochlear hair cells of the mice in model group were lower than those in control group(P<0.05), and the expression levels of GRP78 and caspase-12 were higher than those in control group(P<0.05).Conclusion: ERS can result in the increase of defect rate of cochlear outer hair cells and ABR brainstem hearing threshold of the diabetic mice and decrease the expression levels of p-ERK and Nrf2 proteins, suggesting that ERS can promote the degenerative lesions of cochlear hair cells in the type 2 diabetic mice.

Objective: To investigate the effects of Purendan Superfine Powder(PRD) on the retinal hemodynamics and the expressions of cytokines in the diabetic rats, and to elucidate its protective effect on diabetic retinopathy(DR).Methods: Thirty-six male SD rats were randomly divided into control group, diabetes group, and PRD group(n=12). The diabetes rat models were established by intraperitoneal injecting streptozotocin(STZ) in diabetes group and PRD group. After successful modeling, the diabetic rats in PRD group were given PRD (1.8 g ·kg^-1) by gavage for 3 months. The hemodynamics parameters of central retinal artery(CRA) of the rats were measured with color Doppler ultrasound. The expression levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA in retina tissue were detected by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemical staining and Western blotting methods were used to detect the expression levels of VEGF and bFGF proteins in retina tissue of the rats.Results: Compared with control group, the blood glucose, urine volume, liver index and kidney index of the rats in diabetes group were significantly increased(P<0.05), the peak systolic velocity(PSV), end diastolic velocity(EDV) and mean velocity(MV) of CRA were significantly decreased(P<0.05), the resistant index(RI) and pulsatility index(PI) of CRA were significantly increased(P<0.05), and the expression levels of VEGF and bFGF mRNA and protein in retina tissue were significantly increased(P<0.05). Compared with diabetes group, the blood glucose, urine volume, liver index and kidney index of the rats in PRD group were significantly decreased(P<0.05), the PSV, EDV and MV of CRA were increased(P<0.05), the RI and PI of CRA were decreased(P<0.05), the expression levels of VEGF and bFGF mRNA and protein in retina tissue were significantly reduced(P<0.05).Conclusion: PRD can protect the diabetic retinal microvascular injury by reducing the blood glucose level, increasing the blood supply of retinal tissue, increasing the amount of microcirculation blood perfusion, reducing the vascular bed perfusion resistance, and reducing the expression of VEGF and bFGF.

Objective: To investigate the effects of Jiawei Danshen Yin on the cardiac function and serum inflammatory factors interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in the rats with chronic heart failure, and to elucidate the mechanism of Jiawei Danshen Yin in the treatment of chronic heart failure.Methods: The SD rats were randomly divided into blank control group, heart failure model group, captopril group, and Low, medium,and high doses of Jiawei Danshen Yin groups.The rat models of chronic heart failure were prepared by intraperitoneal injection of adriamycin solution for 4 weeks.The left ventricular ejection fraction (LVEF) of the rats was measured by echocardiography.The heart rate (HR), cardiac output (CO), systolic blood pressure (SBP), carotid artery diastolic pressure (DBP), left ventricular end diastolic pressure (LVEDP), left ventricular systolic pressure (LVSP), left ventricular pressure change maximum increase/decrease rate (±dp/dt_max) and other related hemodynamic parameters were measured by ventilator in each group to assess the rat heart function. The levels of inflammatory cytokines IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).Results: Compared with blank control group, the LVEF of the rats in model group, captopril groups and low, medium, and high doses of Jiawei Danshen Yin groups were decreased (P<0.05). Compared with model group,the LVEF of the rats in captopril group and low, medium, and high doses of Jiawei Danshen Yin groups were increased(P<0.05).The HR, SBP, DBP, CO, LVSP, and ±dp/dt_max of the rats in model group were lower than those in blank control group(P<0.05), while the LVEDP was increased (P<0.05). Compared with model group, the SBP, DBP, CO, LVSP and ±dp/dtmax of the rats in captopril group and low, medium, and high doses of Jiawei Danshen Yin groups were increased(P<0.05),however the HR and LVEDP were decreased (P<0.05). Compared with low dose of Jiawei Danshen Yin group, the CO and ±dp/dt_max of the rats in captopril group and medium and high doses of Jiawei Danshen Yin groups were increased(P<0.05) and the LVEDP were decreased (P<0.05).Compared with blank control group, the levels of serum IL-6 and TNF-α of the rats in the other groups were increased (P<0.05). The levels of serum IL-6 and TNF-α of the rats in captopril group and low, medium, and high doses of Jiawei Danshen Yin groups were lower than those in model group (P<0.05). Compared with low dose of Jiawei Danshen Yin group, the levels of serum IL-6 and TNF-α of the rats in captopril group and medium and high doses of Jiawei Danshen Yin groups were decreased (P<0.05). Compared with blank control group, the indexes of heart and lung in the other groups were all increased (P<0.05); compared with model group, the heart and lung indexes of the rats in captopril group and low, medium, and high doses of Jiawei Danshen Yin groups were decreased (P<0.05).Conclusion: Jiawei Danshen Yin can improve the heart function of the rats with heart failure and inhibit the expressions of serum inflammatory cytokines IL-6 and TNF-α to some extent.In the common dose range, Jiawei Danshen Yin can maximize the benefit of cardiac function and the serum inflammatory factors in the rats with chronic heart failure.

Objective: To investigate the effect of resveratrol (Res) on the fat synthesis in the liver cancer HepG₂ cells, and to elucidate its possible mechanism.Methods: The HepG₂ cells were cultured in vitro and divided into Res group (treated with 40 μmol·L^-1 DMSO- diluted Res for 24 h) and control group (treated with the same concentration of DMSO for 24 h). The cell supernatant was collected, and the levels of triglyceride (TG) and total cholesterol (TC) in the cells in various groups were measured by ELISA. The mRNA and protein expression levels of lipase synthase acetyl-CoA carboxylase (ACC1), fatty acid synthetase (FASN) and stearoyl-CoA desaturase (SCD1) in the cells in various groups were detected by qRT-PCR and Western blotting method. The levels of O-linked N-acetylglucosamine(O-GlcNAc)glycosylation in the cells in various groups were detected by Western blotting method.Results: Compared with control group, the levels of TG and TC in the cells in Res group were decreased, but the difference was not statistically significant (t₁=1.886,P>0.05; t₂=2.457,P>0.05). Compared with control group, the levels of expressions of ACC1, FASN and SCD1 mRNA and proteins in the cells in Res group were significantly decreased (P<0.05 or P<0.01); the O-GlcNAc glycosylation level in the cells in Res group was significantly decreased (t=2.87, P<0.05).Conclusion: Res has the effect of inhibiting the fat synthesis in the liver cancer HepG₂ cells. Its mechanism may be related to the reduction of cellular O-GlcNAc glycosylation level and the reduction of the expression of FASN.

Objective: To observe the expressions of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods: The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7 cells and the malignant glioma LN443 and U343 cells were cultured;the Cos7 cells were used as normal cell control group,and the LN443 and U343 cells as malignant glioma cell group. Western blotting method was used to detect the expression levels of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells.Results: In brain tissue, the expression levels of SENP1, SENP2 and SENP6 proteins in malignant glioma group were higher than those in normal control group(P<0.05). Compared with normal cell control group, the expression levels of SENP1, SENP2 and SENP6 proteins in the LN443 and U343 cells in malignant glioma cell group were significantly increased (P<0.05).Conclusion: SENP1, SENP2 and SENP6 proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.

Objective: To explore the effects of atorvastatin on the natural antisense transcription of apolipoprotein A1 (apoA1-NAT) in the HepG₂ cells and its influence in the expressions of lipid metabolism related genes.Methods: The HepG₂ cells were intervened with different concentrations(0,1,10 and 100 nmol·L^-1) of atorvastatin, and the 0 nmol·L^-1 atorvastatin group was used as control group.The total RNA of HepG₂ cells in various groups were extracted at different time points(6,12,24, and 48 h). The mRNA expression levels of lipid metabolism-related genes and apoA1-NAT expression levels were detected by Real-Time PCR method.Results: The morphology of HepG₂ cells in 1 and 10 nmol·L^-1 atorvastain groups was normal.Compared with 6 h, the expression levels of apoA1-NAT in the HepG₂ cells in 10 nmol·L^-1 atorvastatin group at 12, 24 and 48 h were significantly decreased (P<0.01) in a time-dependent manner. Under the same condition, the expression level of apoA1 mRNA in the HepG₂ cells at 48 h was increased significantly compared with 6 h (P<0.01).Compared with control group, the expression levels of low density lipoprotein receptor(LDLR) in the HepG₂ cells in 100 nmol·L^-1 atorvastatin group,scavenger receptor-class B type 1 (SRB1) in 10 nmol·L^-1 atorvastatin group and ATP binding cassette transporter A1(ABCA1) in 1 and 10 nmol·L^-1 atorvastatin groups were increased(P<0.01) in different degrees.Conclusion: Atorvastatin can promote the expressions of lipid metabolism-related genes by inhibiting the expression of apoA1-NAT, and promote the process of reverse cholesterol transport.

Objective: To explore the differences of wear resistance of three kinds of glass ceramics and Wieland Zenostarzircona (Zenostar),and to clarify their influencing factors.Methods: Zenostar were made into flat-shaped specimens(zirconia base sample group) and hemisphere-shaped specimens (zirconia pair grinding group).There kinds of glass ceramics IPS Empress(Empress),IPS e.max CAD(e.max),VITA Suprinity (Suprinty) were used as base specimens.Each group was exposed to UMT-2 testing machine to simulate the clinical service.The wear depthes of base specimens were detected by laser confocal scanning.Scanning electron microscope(SEM) was used to evaluate the wear surfaces.Results: In zirconia base sample group,there were no significant differences in the maximum wear depthes to Zenostar between the three kinds of glass ceramics(P>0.05).In zirconia pair grinding group, the maximum wear depthes ranked as follows:Zenostar groupP>0.05),but there were significant differences between other groups (P<0.01).The SEM results showed that the wearing surface of the Zenostar in zirconia base sample group was relatively smooth;whereas the wearing surface of Empress in zirconia pair grinding group was rougher with alarge area of clebris desquamation surface.Conclusion: The wear resistance of the three kinds of glass ceramics to Zenostar is related to the compositions and the chemical structures of materials.

Objective: To explore the diagnostic values of Wells score and YEARS algorithm in the patients with pulmonary embolism and to compare the areas under receiver operating characteristic(ROC) curves (AUC) of Wells score and YEARS algorithm, and to find the more suitable score method for pulmonary embolism in clinic.Methods: A total of 139 patients who were suspected with pulmonary embolism were collected, among them 48 patients were clinically as confirmed pulmonary embolism.The disease history,clinical manifestation and results of auxilliary examinations of the patients were collected.All the patients were assessed by Wells score and YEARS algorithm,respectively;the computed tomographic pulmonary angiography(CTPA) was considered as golden standard. The sensitivities, specificities, misdiagnosis rates and omission diagnostic rates,and the accuracies of Wells score and YEARS algorithm of the patients in two groups were analyzed with SPSS 22.0 software;the conformities of results of Wells socre,YEARS algorithm,and CTPA were analyzed.Then ROC curves of Wells score and YEARS algorithm in diagnosis of pulmonary embolism were made,and the AUC was calculated and compared.Results: Compared with YEARS algorithm, the specificity and accuracy of Wells score in diagnosis of pulmonary embolism were markedly increased(P<0.05),the misdiagnosis rate was decreased(P=0.037),however the sensitivity and the misdiagnosis rate had no significant differences(P>0.05).The Kappa value of Wells score and CTPA was 0.45, the conformity was moderate;the Kappa value of YEARS algorithm and CTPA was 0.22,the conformity was passable;the Kappa value of Wells score and YEARS algorithm was 0.11,the conformity was bad.The AUC of Wells score and YEARS algorithm were 0.753±0.044(P<0.01)and 0.585±0.049(P=0.101).Conclusion: The Wells score is superior to the YEARS algorithm in the diagnosis of pulmonary embolism.

Objective: To investigate the expression of activator of basal transcription 1 (ABT1) protein in gastric cancer tissue and its relationships with the clinical parameters and prognosis of gastric cancer patients, and to clarify the role of ABT1 in the occurrence and development of gastric cancer.Methods: A total of 100 cases of cancer tissue of the gastric cancer patients and 80 pairs of adjacent tissue were selected. The expressions of ABT1 in cancer tissue and adjacent tissue were detected by immunohistochemistry and the proportion of stained cells and the degree of staining in the immunohistochemistry results were analyzed using semi-quantitative analysis. The relationships between the semi-quantitative analysis results and the clinical parameters of gastric cancer patients were statistically analyzed. Kaplan-Meier method was used to analyze the correlation between the ABT1 protein expression level and the survival of gastric cancer patients.Results: ABT1-positive staining was observed in the nucleus and cytoplasm of gastric cancer tissue and adjacent gastric tissue. The expression level of ABT1 in gastric cancer tissue was lower than that in adjacent tissue(P=0.021). The ABT1 protein expression level in gastric cancer tissue was significantly negatively correlated with the pathological grade (r=-0.224,P=0.026).The Kaplan-Meier analysis results of the survival curve showed that the high expression of ABT1 was associated with good prognosis in the gastric cancer patients (HR=1.483,P<0.01).The survival rate of gastric cancer patients with high ABT1 expression was significantly higher than that of the patients with low ABT1 expression (HR=2.411,P=0.0272).Conclusion: The expression of ABT1 in gastric cancer tissue is lower,indicating that ABT1 can be used one of the markers of good prognosis of gastric cancer.

Objective: To discuss the curative effect of Chinese medicine synthesis rehabilitation in the treatment of neurogenic bladder dysfunction after incomplete spinal cord injury, and to provide the clinical evidences for its application.Methods: A total of 184 patients meeting the inclusion criteria were randonly divided into trial group (n=89) and control group (n=95) with central area group method. The patients in two groups were given the intermittent catheterization and the urinary functional training. The patients in trial group were treated by the electricity needle (20 min/time, one time a day, 6 d per week) and the massage (20 min/time, one time a day, 6 d per week). All patients in two groups were treated for 4 courses (2 weeks for a course). The residual urine volumes, the bladder securiey capacities,and the intravesical pressures of the patients in two groups were measured before and after treatment,and the curative effects were evaluated.Results: Compared with before treatment the residual urine volume of the patients in trial group was reduced significantly after treatment (P<0.01), the bladder security capacity was significantly increased (P<0.01), and there was no significant change in intravesical pressure (P>0.05); the residual urine volume of the patients in control group was reduced significantly after treatment (P<0.01), there were no significant changes in the bladder security capacity and intravesical pressure (P>0.05).After treatment, the residual urine volume of the patients in trial group was significantly lower than that in control group (P<0.01), while there were no significant differences in the bladder security capacities and intravesical pressures between two groups (P>0.05).Conclusion: Chinese medicine synthesis rehabilitation may reduce the residual urine volume of the patients with neurogenic bladder urinary retention after incomplete spinal cord injury, and its curative effect is superior to modern rehabilitation of intermittent catheterization and urinary bladder function training.

Objective: To analyze the dynamic variety of serum Nesfatin-1 in the patients with status epilepticus(SE), and to explore the clinical values of the serum Nesfatin-1 in evaluation on the illness condition and short-term prognosis in the patients with epileptic seizure.Methods: A total of 43 patients diagnosed as primary epilepsy were collected as the subjects and received the regular clinical therapy. The serum levels of Nesfatin-1 of the patients were detected before treatment and at the end of the 1st week after treatment, the end of the 2nd week therapy after treatment and the end of the 1st month after treatment, respectively. Liverpool Seizure Severity Scale 2.0 (LSSS2.0) was used to evaluate in the patients. The correlations of the serum levels of Nesfatin-1 of the patients at each time point with the scores of LSSS were analyzed.After one year of follow-up, all patients were divided into two groups according to the prognosis, which were survival group (31 cases) and death group (12 cases). The risk factors of one-year prognosis were analyzed by multivariate Logistic regression analysis. The sensitivity and specificity of the serum values of Nesfatin-1 to the one-year prognosis were analyzed by receiver operating characteristic (ROC) curve.Results: Compared with before treatment, the LSSS scores and the serum Nesfatin-1 levels of the patients in two groups were decreased with the increasing of treatment time (P<0.05). The LSSS scores and the serum levels of Nesfatin-1 of the patients were not different between two groups before treatment (P>0.05).The LSSS scores and the serum levels of Nesfatin-1 of the SE patients in survival group were lower than these in death group from the end of the 1st week after treatment to the end of the 1st month after treatment (P<0.05 or P<0.01). The positive correlations of the serum levels of Nesfatin-1 and the LSSS scores of the patients at each time point were confirmed (r=0.617-0.726,P<0.05).The serum high level of Nesfatin-1 was the risk factor of the one-year prognosis analyzed by multivariate Logistic regression analysis. The ROC curve identified the cutoff level of serum Nesfatin-1 (2.7μg·L^-1) to the one-year prognosis with the sensitivity of 88.5% and specificity of 79.6%.Conclusion: The dynamic variety of serum Nesfatin-1 can reflect the condition of illness, and the serum level of Nesfatin-1 is decreased gradually with the prolongation of time, and has application value in predicting the prognosis of the patients with epileptic seizure.

Objective: To investigate the changes of expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in the peripheral blood mononuclear cells and downstream factors interleukin-1β (IL-1β) and interleukin-18 (IL-18) in serum in the children with asthma, and to explore their significances on assessing the condition of the children.Methods: A total of 176 cases of children with asthma were divided into acute exacerbation group (n=91), chronic persistent group (n=49) and clinical remission group (n=36) according to the clinical manifestation. During the same period, 60 healthy children were selected from the outpatient physical examination center as control group.The pulmonary function of children was checked with lung function instrument. The expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and cysteinyl aspartate-specific proteinase-1 (Caspase-1) mRNA in peripheral blood mononuclear cells of the subjects in various groups were detected by using real-time quantitative PCR. The serum levels of IL-1β and IL-18 of the subjects in various groups were detected by using enzyme-linked immunosorbent assay (ELISA).Results: Compared with control group, the forced expiratory volume in 1 second percentage of predicted value (FEV1%) and fixed ratio of forced expiratory volume in the first second/forced vital capacity (FEV1/FVC) of the children in acute exacerbation, chronic persistent and clinical remission groups were decreased (P<0.05);acute exacerbation groupP<0.05).The levels of NLRP3, ASC and Caspase-1 mRNA in the peripheral blood mononuclear cells and serum levels of IL-1β and IL-18 in the children with asthma were higher than those in control group (P<0.01).The expression levels of NLRP3, ASC and Caspase-1 mRNA in the peripheral blood mononuclear cells of the children in acute exacerbation group were higher than those in chronic persistent and clinical remission groups (P<0.05), and the expression levels of NLRP3,ASC and Caspase-1 mRNA in chronic persistent group were higher than those in clinical remission group (P<0.05).Pearson correlation analysis showed that the expression level of NLRP3 mRNA in the peripheral blood mononuclear cells of asthmatic children was positively correlated with the expression levels of ASC, Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC; the expression level of ASC mRNA was positively correlated with the expression level of Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC (P<0.05); the expression level of Caspase-1 mRNA was positively correlated with the expression level of Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC (P<0.05); the serum level of IL-1β was negatively correlated with FEV1% and FEV1/FVC (P<0.05), and the serum level of IL-18 was negatively correlated with FEV1% and FEV1/FVC (P<0.05).Conclusion: The expression levels of NLRP3 inflammasome and the downstream factor IL-1β and IL-18 in peripheral blood of the children with asthma are increased, and they are related to the clinical stage of the children with asthma. NLRP3 inflammasome pathway might promote the pathogenesis of asthma in the children.

Objective: To compare the clinical efficacies of tenofovir(TDF)and entecavir(ETV)in the treatment of the aged patients with chronic hepatitis B(CHB)and their regulation on the inflammation factors,and to provide basis for their clinical application.Methods: A total of 93 aged patients with CHB were selected and randomly divided into TDF group(n=48)and ETV group(n=45). And the patients were treated with TDF (300 mg/time, 1 time·d^-1)and ETV(0.5 mg/time, 1 time·d^-1);the patients in two groups were observed for continuous 48 weeks. The levels of serum hepatitis B virus-deoxyribonucleic acid(HBV-DNA), glutamic pyruvic transaminase(ALT), and tumor necrosis factor α(TNF-α), interleukin-10(IL-10),the ALT recovery rates,the HBV-DNA negative conversion rates,and the HBeAg negative rates of the patients in two groups were detected before treatment and 4,12,24,36 and 48 weeks after treatment, respectively.Results: Compared with before treatment,the levels of serum HBV-DNA and ALT of the patients in two groups after treatment were decreased(P<0.05); the levels of serum HBV-DNA and ALT of the patients in TDF group were significantly lower than those in ETV at 4 and 12 weeks after treatment(P<0.05). Compared with before treatment,the levels of serum TNF-α and IL-10 of the patients in two groups after treatment were significantly decreased, and there were statistically significant differences between two groups at 12 weeks after treatment (P<0.05). After treatment, the ALT recovery rates of the patients in two groups were increased gradually; the ALT recovery rates in TDF group were significantly higher than those in ETV group (P<0.05) at 4 and 12 weeks after treatment. The patients in two groups showed complete virological responses at 4 weeks after treatment, and the HBV-DNA negative conversion rates were increased gradually. The negative conversion rates of HBV-DNA in TDF group were significantly higher than those in ETV group (P<0.05) at 4, 12 and 24 weeks after treatment. The negative conversion of HBeAg in TDF group appeared at 12 weeks, which was earlier than that in ETV group;the negative conversion of HBeAg in ETV group appeared at 24 weeks; there was no significant difference in HBeAg negative conversion rate between two groups (P>0.05).Conclusion: Both TDF and ETV could be used to treat the aged patients with CHB. TDF has earlier antiviral effect and can bring quick recovery in liver function with lighter inflammatory reaction. TDF is an ideal antiviral drug for clinical treatment.

Objective: To explore the curative effects and securities of ticagrelor and clopidogrel in the antiplatelet therapy for coronary heart disease in the aged patients, and to provide a more rational strategy for antiplatelet therapy.Methods: Computer search was performed in Cochrane Central Register of Controlled Trial, Ovid-Medline, EMBase, PubMed, CNKI, Wanfang, VIP and CBM,and the randomized controlled trial (RCT) of ticagrelor and clopidogrel in the treatment of coronary heart disease in the old people were collected. The patients in ticagrelor group were given ticagrelor and other drugs,and the patients in clopidogrel group were given clopidogrel and other drugs. RevMan 5.3 software was used to perform Meta-analysis,and the adverse events related to the effectiveness such as the major adverse cardiovascular events(MACE),myocardial infarction(MI),stent thrombosis,death,stroke and the adverse events related to safety (bleeding event) were observed.Results: According to inclusion and exclusion criteria,a total of 15 RCT were included. The results of Meta-analysis showed that the incidences of MACE(RR=0.59,95%CI:0.46-0.76,Z=4.08,P<0.01), MI(RR=0.48,95%CI:0.28-0.81,Z=2.74,P=0.006), stent thrombosis (RR=0.16,95%CI:0.06-0.48,Z=3.30,P=0.0010),and death (RR=0.52,95%CI:0.30-0.89,Z=2.41,P=0.02) in ticagrelor group were decreased compared with clopidogrel group;there was no significant difference in stroke rate (RR=0.76,95%CI:0.39-1.47,Z=0.81,P=0.42)between two groups,and the incidence of bleeding event in ticagrelor group was increased compared with clopidogrel group(RR=1.57,95%CI:1.20-2.05,Z=3.28,P=0.001).Conclusion: For the aged patients with coronary heart disease,ticagrelor is superior to clopidogrel;but the incidence of bleeding is increased.

Objective: To evaluate the efficacies of clarithromycin and metronidazole in the treatment of chronic periodontitis,and to provide the evidence-based medical evidence for the rational use of drugs in the treatment of chronic periodontitis.Methods: CNKI, VIP, Wanfang, Chinese Biomedical Literature Database, PubMed, ScienceDirect and EMbase database from inception to June, 2017 were searched by computer for the literatures about the treatment of clarithromycin and metronidazole for chronic periodontitis. Two reviewers independently screened the literatures, extracted the data and evaluated the bias risk of included studies. Then Meta-analysis was performed using RevMan 5.3 software.Results: A total of 5 randomized controlled trial(RCT)involving 516 patients with chronic periodontitis were included. Subgroup analysis was performed according to the follow-up time. Compared with metronidazole group,the probing depth(PD) reduction,attachment loss (AL) gain,and sulcus bleeding index (SBI) reduction of the patients with chronic periodontitis in clarithromycin group at 1 month and 3 months after follow-up were more significant;the differences in PD, AL, and SBI were significant(MD=-0.53, 95%CI:-0.67-0.39, P<0.01;MD=-0.31, 95% CI:-0.39—-0.24, P<0.01;MD=-0.23, 95% CI:-0.29—-0.16, P<0.01).Conclusion: Systemic antibiotic use of clarithromycin in the treatment of the patients with chronic periodontitis after non-surgical periodontal therapy has a significant additional effect than metronidazole in short-term observation.

Objective: To investigate the relationships between the expression of insulin-like growth factor-1 (IGF-1) in the gastric cancer tissue and the clinicopathological parameters in the gastric cancer patients in China with Meta-analysis, and to provide the evidence-based medical evidence for the diagnosis and treatment of gastric cancer.Methods: The literatures about the relationships between the positive expression rate of IGF-1 and the clinicopathological paramters in the gastric cancer patients were searched comprehensively in PubMed,China National Knowledge Infrastructure, Weipu Chinese Science and Technology Journal Database, Wanfang Database and China Biomedical Literature Database. Review Manager 5.2 software was used to conduct Meta analysis on the literatures that met the inclusion criteria.Results: There were 11 articles in the literatures were selected, including 993 cases in gastric cancer group and 601 cases in non-tumor gastric mucosal tissue control group. Meta-analysis showed the significant differences in the positive expression rates of IGF-1 between gastric cancer group and control group (OR=9.05, 95% CI:6.82-12.01), poorly differentiated gastric cancer group and moderately-highly differentiated group (OR=1.53, 95% CI:1.14-2.06), involving full-thickness group and unaffected full-thickness group (OR=0.16, 95% CI:0.09-0.28), lymph node metastasis group and non-lymph node metastasis group (OR=0.12, 95% CI:0.08-0.17),TNM stage Ⅲ-Ⅳ group and TNM stage Ⅰ-Ⅱ group (OR=0.21, 95% CI:0.13-0.33).Conclusion: The high expression of IGF-1 in gastric cancer tissue is closely related to the occurrence, development, invasion and metastasis of tumor in the gastric cancer patients in China. The abnormal expression of IGF-1 could be used as a risk factor for carcinogenesis of gastric mucosal cells and a valuable molecular biological marker for early diagnosis and treatment of gastric cancer.

Objective: To explore the change in the cerebral natriuretic peptide (BNP) levels in the premature infants with hemodynamically significant patent dectus arteriosus (hsPDA) and its relationship with hemodynamics, and to further clarify the clinical significance of BNP in the judgement of illness condition and treatment in the premature infants with hsPDA.Methods: A total of 106 cases of premature infants within 6 h of birth whose gestational age was no more than 32 weeks were selected and divided into hsPDA group (43 cases) and PDA with no hemodynamic significance group (nhsPDA group, 27 cases) and no PDA group (nPDA group, 36 cases).The patients in HsPDA group were divided into hsPDA treatment group(33 cases) and hsPDA non-treatment group(10 cases) according to whether ibuprofen was administrated or not. Echocardiography was performed in the patients in hsPDA treatment group,7 d after oral ibuprofen administration, and the patients in hsPDA treatment group were divided into hsPDA close group(15 cases) and hsPDA non-close group(18cases). The left atrium (LA)/aortic root diameter (VO) value, diameter of patent arterial duct, left ventricular ejection fraction(LVEF), shortening fraction, left ventricular end-diastolic diameter(LVEDD) and left ventricular end-systolic diameter(LVESD) were recorded at 1, 3, 7 d after birth of the premature infants in various groups with echocardiography.The level of BNP in plasma was determined by electrochemiluminescence. The correlation analysis was performed between the hemodynamic indexes of echocardiography and the level of BNP in plasma.Results: Compared with nhsPDA group, the LA/VO value, arterial catheter diameter and LVEDD were significantly increased (P<0.05).The plasma BNP levels of premature infants in hsPDA close and non-close groups at 1 and 3 d after birth were significantly higher than those in nPDA group (P<0.05); the plasma BNP level of the premature infants in hsPDA close group at 7 d after birth was significantly lower than that at 3 d after birth (P<0.05).Compared with hsPDA non-treatment group, the LA/VO value, arterial catheter diameter,LEVDD and the plasma BNP level of the premature infants in hsPDA treatment group at 7 d after birth were significantly decreased (P<0.05). The plasma BNP level was positively correlated with the LA/VO value and arterial catheter diameter (r=0.727,P<0.05;r=0.780, P<0.05) of the premature infants in hsPDA group at 3 d after birth.Conclusion: The level of plasma BNP of premature infants 3 d after birth is positively correlated with the hemodynamic indexes, and the detection of the changes of the plasma BNP levels is helpful to judge the condition of preterm infants with hsPDA and to provide basis for its diagnosis.

Objective: To discuss the clinical characteristicsof the Pneumocystis jirovecii pneumonia (PCP) in the non-HIV-infected blood disease patients,and to analyze its risk factors, treatment methods, prognosis and prevention measures.Methods: A female patient aged 18 years old was confirmed as acute myeloid leukemia (AML),and experienced dyspnea, chest congestion and hypoxaemia during the recovery period of hemogram after chemotherapy. The chest CT showed the bilateral lung diffuse ground glass density images. The patient had a dry cough and the oxygen saturation was gradually decreased to 75% 5 d after antibacteriological treatment. A repeat chest CT showed enlarged diffuse ground glass density images on both lungs. Considering about the possibility of PCP,the patient received oral trimethoprim/sulfamethoxazole(TMP/SMX) 1 g, once every 6 h, in combination with caspofungin.Results: Two days later,the symptoms of the patients were not improved. The patient was transferred to ICU and was diagnosed PCP by bronchoalveolar lavage. The patient was switched to oral TMP/SMX 2g, once every 8 h, in combination with caspofungin. Meanwhile,the patient received bi-level positive airway pressure ventilation (Bipap) for the increased work of breathing. Five days later,the symptoms of the patients were improved and the Bipap was stopped. The patient got better and discharged 5 d later. The patient continuely received oral TMP/SMX 2 g, once every 8 h for 36 d.Conclusion: Prevention of PCP should be focused, in the non-HIV-infected blood disease patients receiving chemotherapy. Diagnosis of PCP should be considered in these patients without prevention who once have suspected clinical manifestation of PCP in non-granulocytic phase. Early empirical treatment of PCP and ICU management in the non-HIV-infected blood disease patients with acute respiratory failure are the keys to reduce death and improve the prognosis of PCP.

Objective: To discuss the clinic features and diagnosis of one case of hepatic angiomyolipoma(HAML), and to analyze and summarize its diagnosis and treatment methods.Methods: The clinical materials of one case of HAML patient who was misdiagnosised as hepatocellcular carcinoma were analyzed retrospectively,and the related literatures were reviewed.Results: A middle-aged female patient visited our outpatient service because of low back discomfort.The upper abdominal CT and MRI suggested that a space occupying lesion located at the left lobe of liver inner lower segment,considering a high possibility of hepatocellular carcinoma.The patient underwent the left lobe of inner lower segmentectomy.The postoperative histopathological examination and immunohistochemical staining showed the tumour was HAML,After operation,the patient recovered well and the follow-up examination after hepatectomy found no tumor recurrence.Conclusion: HAML has no specific clinical characteristics or imaging features.The diagnosis of HAML mostly depend on the histopathological examination and hepatectomy is the effective treatment.

Objective: To study the pathogenesis and the risk factors of eye injury after non-ocular surgery in the patients underwent general anesthesia, and to provide the reference for its prevention,diagnosis and treatment.Methods: The clinical materials of two patients with eye injury after non-ocular surgery underwent general anesthesia were analyzed and the related literatures were reviewed.Results: A young woman patient underwent laparoscopic hysterectomy with general anesthesia while positioned Trendelenburg,the eyes were being closed naturally without protection,and corneal abrasion of both eyes occurred after operation. The lesion had completely resolved with no sequelae after treatment.An old man underwent cervical posterior laminoplasty with general anesthesia in prone position developed ischemic optic neuropathy(ION) after operation. The vision of the patients partly recovered after symptomatic treatment.Conclusion: Corneal abrasion is the most frequent ophthalmologic complication during general anesthesia,most of the patients have good prognosis. Postoperative visual loss(POVL) is the most severe ophthalmologic complication without effective treatment available,the delicate reasons and mechanisms are not totally clear,prevention outweighs treatment.

Objective: To investigate the efficacy of local application of methimazole cream in the patients with hyperthyroidism complicated with liver failure, and to provide reference for its clinical treatment.Methods: Two patients were admitted to hospital with jaundice as the main performance. Liver function and coagulation function supported the diagnosis of liver failure. The thyroid function indicators suggested hyperthyroidism. Combined with other related examination results, liver failure, hyperthyroidism and hepatitis B virus were diagnosed,and the local treatment of methimazole cream and other related treatments were given.Results: After treatment, the liver function of two patients returned to normal and the thyroid function was improved. After follow-up, one patient changed the external use of methimazole cream to oral treatment,and another patient was still treated in a local manner.Conclusion: There is a conflict in the treatment of hyperthyroidism complicated with liver failure. The local application of methimazole cream in the topic of reducing the risk of liver damage caused by drugs has good results in the treatment of hyperthyroidism. This method also provides the new ideas for its clinical treatment.

Objective: To study the CT findings of hepatic lesions without obvious enhancement in arterial phase, and to analyze the characteristics of the hepatic lesions.Methods: The clinical materials of 118 cases of hepatic lesions without obvious enhancement in arterial phase scanned by enhanced CT were collected. The basic CT signs, characteristic CT signs and enhancement patterns were analyzed.Results: Among 118 cases, there were 40 hepatic metastases, 29 liver abscesses, 21 intrahepatic cholangiocelloma, 17 hepatocellular carcinoma, 6 inflammatory pseudotumor of liver and 5 hepatic lymphoma; 54 cases showed single lesion and 64 cases showed multiple lesions; 24 cases were sharp and 94 cases were indistinct; 41 cases located in right hepatic lobe, 26 cases located in left hepatic lobe, 50 cases located in left and right hepatic lobe, and 1 case located in hepatic caudate lobe. The diameters of lesions were 0.2-13.4 cm. 40 cases of hepatic metastases showed mild marginal enhancement, while the incidence rate of bull eye sign was 12.50% (5/40); 29 cases of liver abscess showed ring or honeycomb-like enhancement, while the incidence rates of double ring sign and gas bubbles were 13.79% (4/29) and 13.79% (4/29).Seventeen cases of intrahepatic cholangiocelloma showed slow progressive enhancement,while the incidence rates of intrahepatic bile duct dilatation, hepatic capsular depression and calcification were 57.14% (12/21), 23.81% (5/21) and 9.52%(2/21). Seventeen cases of hepatocellular carcinoma showed various enhancement patterns, while the incidence rates of liver contour change, hepatic artery enlargement and vessel invasion were 41.18% (7/17), 41.18% (7/17) and 35.29%(6/17). Four cases of inflammatory pseudotumor of liver showed septal and nodular enhancement; 5 cases of hepatic lymphoma showed internal minimal homogeneous enhancement, while the incidence rate of ‘vascular floating’ sign was 40.00% (2/5).Conclusion: Spiral CT examination of the hepatic lesions without obvious enhancement in arterial phase should be combined with CT signs and enhancement patterns for synthetic judgment, which is helpful to provide an important basis for the qualitative and differential diagnosis of liver lesions.

Objective: To analyze the relationship of hepatic transit time and portal vein pressure in the patients with portal hypertension and esophagogastric varices, and to clarify its clinical significance.Methods: A total of 50 hospitalized patients underwent esophagogastric venous embolization due to portal hypertension and esophagogastric varices were selected as observation group, and 50 patients without history of liver disease and clinical manifestations of liver disease (hospitalized or out-patient) were selected as control group, All patients underwent liver contrast enhanced ultrasound.The free portal pressure (FPP)was measured by ultrasound guided portal vein puncture for operating pathways in the patients in observation group. The hepatic artery-hepatic vein transit time (HA-HVTT), the hepatic portal vein-hepatic vein transit time (PV-HVTT), the liver parenchyma-hepatic vein transit time (PA-HVTT) of the patients were compared between two groups. The correlations between HA-HVTT, PV-HVTT, PA-HVTT and FPP of the patients in observation group were analyzed.Results: The HA-HVTT, PV-HVTT and PA-HVTT of the patients in observation group were significantly shorter than those in control group(t=5.078,P<0.01;t=12.163,P<0.01;t=2.649,P<0.01).The HA-HVTT, PV-HVTT and PA-HVTT had negative correlations with FPP in observation group(r=-0.799,P<0.01;r=-0.554,P<0.01;r=-0.735, P<0.01).The linear relationship between HA-HVTT and FPP was Y=-0.410X+7.254; the linear relationship between PV-HVTT and FPP was Y=-0.355X+4.983; the linear relationship between PA-HVTT and FPP was Y=-0.566X+4.997.Conclusion: Liver transit time can be used as an effective index to judge the portal vein pressure, and it can provide the theroretical basis for the diagnosis and treatment of portal hypertension in the patients.

Objective: To detect the blood flow sepectrum of fetal ductus venosus (DV) with colorDoppler ultrasound, and to explore the application of the fetal DV in screening the chromosome abnormalities.Methods: The blood flow spectra of fetal DV in antenatal examination normal group (320 cases, with 11-40 weeks of gestation) and antenatal examination abnormal group (48 cases, within 11-35 weeks of gestation) were detected with color Doppler and pulse Doppler ultrasound, and chromosome examination was performed in abnormal group. The fetal DV blood flow patterns (S wave, D wave, a wave) were observed, and karyotype analysis was performed in abnormal group.Results: The normal Doppler spectrum of the DV in fetuses showed the same three-phase waves while the abnormal Doppler spectrum of the DV in fetuses showed a wave reserved or absented.A total of 48 cases of chromosome in abnormal group were examined; 12 cases of chromosome abnormality were found in this study (10 cases of DV abnormality and 2 cases of DV normality).A total of 36 cases of normal chromosomes were found which (11 cases of abnormal DV and 25 cases of normal DV).The abnormal detection rate of DV blood flow spectrum in chromosome abnormal group was higher than that in chromosome normal group (χ²=9.723,P=0.002).Conclusion: The abnormality of fetal DV blood flow spectrum can increase the detection rate of chromosome abnormalities and ultrasonography could be used as the primary basis for screening the chromosome abnormality in early pregnancy.

Objective: To investigate the metal artifact reduction effect of orthopedic metal artifact reduction (O-MAR) techonology and its improvement effect on the image quality in CT examination in the patients with arthroplasty,and to elaborate the significance of the technology in clinical diagnosis and treatment of arthroplasty.Methods: The CT data of 20 patients with hip or knee prostheses was collected. There were two tube voltages in each group of CT data:120 and 140 Kev. There were also two groups of CT data in each tube voltage group:non-O-MAR group and O-MAR group;there were four subgroups of CT data of each case:120 Kev/-O-MAR, 120 Kev/+O-MAR, 140 Kev/-O-MAR, 140 Kev/+O-MAR. After data collection, Mimics software was applied to conduct three-dimensional(3D) reconstruction for purpose of the qualitative and quantitative analysis of CT data. Qualitative analysis mainly included the grade of severity of metal artifact and quality of data. Quantitative analysis included the volume of metal artifact,the average CT value and standard deviation(SD) in region of interest (ROI). ROI 1 and ROI 2 were chosen at the location of beam hardening artifact (radial high-density metal artifact) and photon starvation artifact (band low-density metal artifact), respectively.Results: According to the result of 3D measurement, the volumes of artifact had no significant difference between 120 Kev/-O-MAR group and 140 Kev/-O-MAR group(P=0.062),but there were siginificant differences in the volumes of artifact between other groups(P<0.05); O-MAR technology decreased the volume of beam-hardening artifact obviously(P<0.05). According to the results of two-dimensional (2D) measurement,there was no significant difference in the average CT values in ROI 2 between 120 Kev/-O-MAR group and 140 Kev/-O-MAR group(P=0.069),but there were significant differences in the average CT values between other groups(P<0.05);O-MAR technology decreased the high-density beam-hardening metal artifact and the low-density photon-starvation metal artifact in 2D measurement.Conclusion: O-MAR technology could significantly reduce the CT metal artifact of hip and knee prostheses and increase the clinical value of CT data.

Objective: To construct and identify the monoclonal antibody of ATP synthase beta subunit(ATP5B) with high purity, and to lay foundation for further study.Methods: The ATP5B gene was amplified by PCR and cloned into the pET28a vector and transformed into E. coli BL21 (DE3). The protein expression was induced by IPTG and then the fusion protein was purified by nickel affinity chromatography column. The protein purity was detected by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Three female Balb/C mice were immunized with purified fusion protein and the tail vein blood was taken to detect the titer of ATP5B antibody by indirect enzyme-linked immunosorbent assay (ELISA). The spleen cells from the immunized mice with the highest serum titer were mixed with the SP2/0 cells to establish the hybridoma cells and the fused cells were screened by indirect ELISA and monoclonally cultured. Karyotype analysis were performed in the positive cells. The hybridoma cells were intraperitoneally injected into 12 weeks old BALB/C mice to estabilish the ascites models. The titer of ascites was detected by indirect ELISA. The purity of the antibody was detected by SDS-PAGE. The antibody subtype was detected by ELISA.Results: After PCR amplification, a specific band of 1 455bp was obtained, and the pET28a empty vector was ligated to obtain a recombinant pET28a/ATP5B vector. The target protein was expressed in the IPTG-induced bacteria solution; the SDS-PAGE results showed that the protein band was found at 51 000. The indirect ELISA results showed that the serum titer of the venous blood of immunized mice was up to 1:64 000. In karyotype analysis, the total number of chromosomes in hybridoma cells was about the sum of myeloma cells and normal mouse spleen cells. The mouse ascites was prepared with the hybridoma cell line, and the highest titer of the antibody was 1:240 000. The subtype of the monoclonal antibody produced by the hybridoma cells was IgG1.Conclusion: The monoclonal antibody against ATP5B protein is successfully prepared by cloning, expressing and purifying the recombinant protein.

Objective: To prepare the lithium-doped poly-glycerol sebacate (PGS-Li) scaffold using the specific effects of lithium ions and the excellent performance of PGS, and to provide the basis for its application prospects in cementation tissue engineeringscaffold.Methods: The scaffolds were divided into two groups. The PGS-Li scaffolds prepared by adding lithium phosphate during the PGS cross-linking process were used as PGS-Li group, and the PGS scaffolds synthesized by the equal-purification of sebacic acid and glycerol were used as PGS group. The molecular weights of the scaffolds in two groups were determined by gel permeation chromatography. The structures of the scaffolds in two groups were analyzed by fourier transform infrared spectroscope. The surface morphology and the porosities and the pore sizes of the scaffolds in two groups were observed by scanning electron microscope. X-ray photoelectron(XPS) spectroscope and inductively coupled plasma optical emission spectrometer were used to determine the Li ion contents in the scaffolds in two groups. Thermogravimetric analyzer was used to analyze the thermal stabilities of the scaffolds in two groups. Contact angle measuring instrument was used to compare the hydrophilicities of the scaffolds in two groups. In vitro weight loss test was used to determine the degradation rates of the scaffolds in two groups. The OCCM-30 cells were divided into experimental group (added with PGS-Li scaffold extract), PGS group (added with PGS scaffold extract) and blank control group (added with DMEM culture medium). MTT assay was used to detect the proliferation activities of cells in various groups at different time (24, 48 and 72 h);the cell morphology was observed by calcein-AM staining.Results: The gel permeation chromatography results showed that the molecular weight of the PGS-Li scaffold was slightly larger than that of the PGS scaffold. The specific absorption peak of phosphate was detected in the fourier infrared spectrum of the PGS-Li scaffold. The scaffolds in two groups had irregular three-dimensional network structures under scanning electron microscope,and the pore size was 20-160 μm, the porosity of PGS scaffold was (53.92±2.18)%, and the porosity of PGS-Li scaffold was (53.58±1.73)%, there was no statistical difference between two groups (P>0.05). The XPS results showed that a peak appeared at 54.9 eV in PGS-Li group, which coincided with the Li 1s binding energy, while the inductively coupled plasma emission spectrometer results showed that the Li ion content in the PGS-Li scaffold was 0.084%. The thermogravimetric analysis results showed that PGS-Li scaffolds began to degrade at a higher temperature and ceased at a lower temperature compared with PGS scaffolds. The contact angle measurement results indicated that both the materials were hydrophilic materials; the contact angle of PGS scaffold meterial was 78.26°±2.00°, and the contact angle of the PGS-Li scaffold material was 69.78°±1.15°; there was statistical difference between two groups (P<0.05). The in vitro degradation experiments showed that the degradation rate of PGS-Li scaffolds was faster than that of PGS scaffolds.The proliferation activity of OCCM-30 cells in PGS-Li group had no significant difference compared with PGS group and blank control group(P>0.05). The calcein-AM staining results showed the green fluorescence in the OCCM-30 cells in PGS and PGS-Li groups,and there were no significant changes in the morphology of cementoblasts.Conclusion: PGS-Li scaffolds have similar composition and structure to PGS scaffolds, and have better performance in hydrophilicity and thermal stability. PGS-Li scaffolds have no effect on the proliferation of cementoblasts and have broad application prospects in cementum tissue engineering.