Abstract:Objective：In this study，TNF-α treated mouse preosteoblast cell line MC3T3-E1 was used to investigate the effect of TNF- treated mouse preosteoblast cell line MC3T3-E1 on its hematopoiesis and its possible mechanism（s）. Methods：MC3T3-E1 cells pretreated with TNF-α were used as feeder layer cells，combined with hematopoietic growth factor SCF，TPO and Flt3-ligand，and Sca-1（+）cells of mice were amplified and sorted in vitro for 7 days，and the granulocyte - single-line colony formation unit（CFU-GM），red line formation unit（BFU-E）and pre-b line formation unit（CFU-pre-B）were detected. The effect of Erk inhibitor FR180204 on the levels of N-cadherin and p-Erk in MC3T3-E1 cells was also examined. Results：After pretreatment with TNF-α，the expression of N-cadherin was up-regulated，p-Erk was down-regulated，and p-Akt had no effect. The feeding layer cells combined with hematopoietic growth factor were amplified into Sca-1（+）cells in vitro for 7 days，compared with the control group，the number of total CFU，BFU-E and CFU-GM in TNF-α pretreatment group decreased（P < 0.05），but the number of CFU-pre-B increased significantly（P < 0.05）. Erk inhibitor FR180204 treated MC3T3-E1 cells，it showed down-regulated Erk and up-regulated N-cadherin levels. Conclusion：TNF-α may indirectly affect production of the blood and differentiation of hematopoietic stem cells into various lines by affecting hematopoietic supporting cells in bone marrow microenvironment. Regulation of the phosphorylation of Erk1/2 and the expression of N-cadherin in osteoblasts may be one of the mechanisms by which TNF-α affects bone marrow hematopoiesis

Abstract:Objective：This study was designed to explore the brain insulin signaling and the expression of glucose transporters（GLUTs）in APP/SP1 transgenic Alzheimer’s disease（AD）mice，it may provide the evidence for the early diagnosis of AD. Methods：Western blot was used to detect the expressions of insulin-signaling pathway related proteins in the cortex and hippocampus of the APP/PS1 transgenic AD model mice. Results：Our results showed that in the 3-month-old mice，brain insulin signaling was irritably activated，and the phosphorylation level of downstream AKT/GSK3β and other insulin signaling molecules increased，while the expression of GLUTs did not change significantly at this time. However，in the 5-month-old mice，it showed that the phosphorylation level of the brain insulin signal decreased significantly，and the phosphorylation level of its downstream AKT/GSK3β and other signaling molecules was down-regulated. At the same time，down-regulated expression of GLUT3 and GLUT4 was observed in the hippocampus，and the level of Tau protein phosphorylation（p-tau）was significantly increased. Conclusion：Our results confirmed that in APP/PS1 transgenic mice，brain insulin signaling pathway and glucose homeostasis were significantly disrupted in the process of AD formation，and the impairments increase with age.

Abstract:Objective：To investigate the effects of omega-3 polyunsaturated fatty acids（n-3 PUFAs）on myocardial extracellular matrix remodeling in rats with myocardial hypertrophy after abdominal aortic coarctation（AAC）. Methods：Forty male Sprague-Dawley rats were randomly divided into sham operation group（group A）and model group. AAC was performed in model group. One week after operation，the model group was randomly divided into three groups：AAC group（group B），AAC + n-3PUFAs low dose group（group C），AAC+ n-3PUFAs high dose group（group D）. The group C and group D were given by gavage 400 mg/（kg·d）and 1 000 mg/（kg·d）n-3 PUFAs respectively，and the group A and group B were given an equal volume of normal saline. After 8 weeks，the rats were sacrificed，and the left ventricular mass index（LVW/BW）and cardiac mass index（HW/BW）were measured. The myocardial tissues were stained with HE and Masson. The serum hydroxyproline was determined by alkaline hydrolysis method. The content of matrix metalloproteinase-9（MMP-9），tissue inhibitor of metalloproteinase-1（TIMP-1）and fibronectin（FN）in myocardial tissue were analyzed by Western blot. Results：Compared with the group A，LVW/BW，HW/BW，and hydroxyproline content in groups B，C，and D increased significantly（P < 0.05）. Pathological staining showed cardiomyocyte hypertrophy，collagen fiber and MMP-9 expression increasing. The expression of TIMP-1 decreased（P < 0.05）. The expressions of FN in groups B，C and D were significantly higher than that in group A（P < 0.05）. Compared with group B，LVW/BW，HW/BW，and hydroxyproline content in group C and group D decreased，the myocardial cells shown less hypertrophy by pathological staining and the collagen content also decreased，and MMP-9 and FN expression decreased，whereas TIMP-1 expression increased（P < 0.05）. Compared with group C，the changes of all the above indexes in group D were greater（P < 0.05）. All these differences have statistical significance. Conclusion：n-3PUFAs have protective effects on myocardial hypertrophy after abdominal aortic coarctation. The mechanism may be related to the inhibition of overexpression of MMP-9 and FN，the up-regulation of TIMP-1 expression，and the influence on the changes in extracellular matrix components.

Abstract:Objective：To investigate the function of oncogene c-Src in the process of anti-estrogen receptor α therapy and how it mediates the resistance in estrogen receptor（ER）-positive breast cancer cells. Methods：Wild-type MCF-7 cells were long-term treated with tamoxifen（TAM） and established tamoxifen resistant cells（TAM-R）. The expression of c-Src，ERα，and epithelial growth factor receptor（EGFR） was detected by immunoblotting. The interactions between ERα and EGFR were measured by immunoprecipitation. The c-Src inhibitor PP2 was used to block the tyrosine kinase activity of c-Src. Results：Compared with wild-type MCF-7 cells，expression levels of ERα，EGFR，and c-Src were not altered in TAM-R cells. However，the phosphorylation of c-Src was increased in TAM-R cells. Further examination demonstrated that interaction between ERα and EGFR was increased in TAM-R cells. Blocking c-Src phosphorylation by PP2 dissociated the interaction between ERα and EGFR in TAM-R cells. Importantly， TAM could onceagain remarkably inhibit cell growth of TAM-R cells after treated by PP2. Thus，the c-Src inhibitor could reverse TAM-R cells to TAM-sensitive cells. Conclusion：Our results suggested that c-Src is a critical molecule to mediate tamoxifen resistance in breast cancer cells through increasing the interaction between ERα and EGFR. Blocking interaction between ERα and EGFR by PP2 can recover the sensitivity to TAM in resistant cells. All of these findings demonstrated that the c-Src inhibitor can be alternatively used with ERα target therapy to treat ER-positive breast cancer thereby improving the therapeutic effects on breast cancer patients.

Abstract:Objective：The present study aims to determine whether pharmacological activation of the transcriptional coactivator with PDZ-binding motif Tafazzin（TAZ） by chemical TM-25659 can promote osteogenic differentiation and bone formation of adipose-derived stem cells（ADSCs）in vitro and in vivo. Methods：Human ADSCs were isolated and expanded in vitro. The changing expressions of TAZ during ADSCs differentiation at different time points were measured by Western blot and real-time quantitative reverse transcription polymerase chain reaction（PCR）. Alizarin red S and oil red O staining were used to detect osteogenic and adipogenic effects. The ADSCs following initial osteogenic differentiation and TM-25659 were loaded on scaffold β-tricalcium phosphate（β-TCP）and then subcutaneously implanted into nude mice. Six weeks later，the results of bone formation in vivo after implantation of the ADSCs were measured and compared by Hematoxylin-Eosin staining，Masson staining and immunohistochemistry staining. Results：TAZ was upregulated during osteogenic differentiation of ADSCs but downregulated during adipogenic differentiation. Pharmacological activation of TAZ by TM-25659 promotes osteogenic differentiation of ADSCs in vitro. The transient treatment of ADSCs with TM-25659 significantly enhances new bone regeneration of ADSCs in vivo. Conclusion：TAZ is a key mediator for promoting osteogenic differentiation of ADSCs. The pharmacological activation of TAZ in ADSCs might become a feasible treatment for bone regeneration and repair.

Abstract:Objective：To establish a stable and reliable model of zebrafish hyperlipidemia and evaluate the effect of high cholesterol diet on the lipid metabolism in zebrafish larvae. Methods：Zebrafish larvae after five days post fertilization（5dpf）were randomly divided into two groups：normal feed group with dry powder of pure egg yolk，and high cholesterol feed model group with different percentages of cholesterol added to pure egg yolk. After 10 days，the evaluation was carried out by microscopy and detecting the physiological and biochemical indicators in the zebrafish larvae. Results：Confocal microscopy showed that the lipid deposition in caudal artery of juvenile fishes in high cholesterol diet model group was significantly higher than that in common diet group（P < 0.05），and the lipid deposition in caudal artery of juvenile fishes increased with the increase of cholesterol content in diet. When the feed intake of the 5% high cholesterol diet group increased，the lipid deposition in blood vessel increased significantly at the beginning（P < 0.05）and subsequently stabilized gradually，but the survival rate of juvenile fishes decreased significantly（P < 0.05）. The body width of fish larvae in the 5% high cholesterol diet group was significantly larger than that in common diet group（P < 0.01），but no significant differences in body length were observed between the two groups（P > 0.05）. The levels of total cholesterol（TC）and total triglyceride（TG）in juvenile fishes fed with 5% high cholesterol diet were both significantly higher than those fed with normal diet（P < 0.01），and the levels of LDL-C were also significantly higher（P <0.05）. Conclusion：Feeding 5% high cholesterol diet for 10 days could induce the occurrence of hyperlipidemia in zebrafish. Increasing the percentage of cholesterol in diet or increasing the dosage of high cholesterol properly could accelerate the establishment of hyperlipidemia model.

Abstract:Objective：To investigate the prevalence of osteoporosis and the changes of bone metabolic markers in patients with connective tissue disease（CTD）by measuring bone mineral density and bone metabolism index，and evaluate the effect of YunKe on the patients with rheumatoid arthritis（RA）. Methods：In the study，280 patients with connective tissue disease in the Affiliated Hospital of Jiangsu University from February 2016 to February 2017 were recruited. Among them，there were 75 cases of systemic lupus erythematosus（SLE），45 cases of primary Sjogren syndrome（pSS）and 160 cases of RA. Bone mineral density（BMD）of lumbar spine L1-L4 and the proximal femur were measured using dual energy X-ray absorptiometry（DXA）in all patients. The bone metabolic markers，serum osteocalcin（OC），N-terminal propeptide of type Ⅰ procollagen（PINP）and C-terminal cross-linking telopeptide of type I collagen（CTX）were also detected by electro chemiluminescence immunoassay. The bone metabolism index OC，PINP and CTX were analyzed respectively with age，duration，body mass index（BMI），C-reactive protein（CRP）and erythrocyte deposition rate（ESR），complement C3，rheumatoid factor（RF），bone density，and hormone usage. Seventy RA patients were treated with YunKe for 4 months and the effect of YunKe on the bone metabolic markers was evaluated. Results：A total of 280 patients had a mean age of（53.96 ± 14.10）years，mean duration of（94.88 ± 108.40）months and mean BMI of（21.84 ± 2.27）kg/m2，in which 27.7% patients showed osteopenia and 21.5% patients showed osteoporosis. Compared with the SLE patients，OC，PINP and CTX levels gradually increased in pSS and RA patients. OC，PINP and CTX levels in pSS and RA patients also gradually increased in the osteopenia group and the osteoporosis group. There were significant differences between normal bone mass group and osteoporosis group（P < 0.05）. OC，PINP and CTX levels in pSS were positively correlated with serum RF（OC：r1=0.570，P=0.002；PINP：r2=0.752，P=0.000；CTX：r3=0.660，P=0.000）；OC and PINP levels in SLE were negatively correlated with BMD of hip（OC：r1=-0.382，P=0.028；PINP：r2=-0.527，P=0.002）；OC and CTX levels in pSS were negatively correlated with BMD of hip（OC：r1=-0.471，P=0.013；CTX：r3=-0.422，P=0.028）. The level of OC was significantly increased（P < 0.05）and the level of ESR was significantly decreased（P < 0.05）after treated with YunKe. The differences of OC and ESR were statistically significant，while the levels of PINP，CTX，CRP and BMD were no significant differences between before and after 4 months of the treatment. Conclusion：Osteoporosis in connective tissue disease patients is common and there are different degrees of bone metabolism changes. Bone metabolic markers can probably predict bone loss in the early stage. YunKe has good effect on bone metabolism for RA.

Abstract:Objective：To investigate the clinical characters of a large family with non-syndromic hereditary hearing loss，and to find the mutational genes. Methods：Clinical and audiological examinations were performed to rule out syndromic hearing impairment，and the inheritance mode of the family was evaluated. The known deafness-associated genes were sequenced by targeted next-generation sequencing. Results：The family has 97 members in 6 generations，of whom 18 persons were affected. The mode of inheritance should be autosomal dominant according to the pedigree. Audiograms showed the Ⅲ4，Ⅳ20，Ⅲ2 and his offspring of this family were late-onset，progressive non-syndromic sensorineural hearing loss. The age of onset was about 28-year-old. V10 and V35 exhibit hearing impairment in high frequencies. Ⅲ4 and her children（V36，V37，V38，V39）showed non-progressive hearing impairment at 3 and 4 year-old. We did not find any known deafness-associated gene mutations by target sequence capture sequencing technology except mitochondrial A1555G mutation in Ⅲ4 and her children（V36，V37，V38，V39）. Conclusion：Pedigree analysis showed an autosomal dominant hereditary pattern in this family. Hearing loss was congenital，bilateral symmetric，and sensorineural. The known deafness genes seem not contribute to the pathogenesis of the hearing loss in this family，suggesting new gene（s） involvement.

Abstract:Objectives：To assess the airway function of the patients with interstitial lung disease（ILD）using the conventional lung function test and the impulse oscillometry system（IOS）. Methods：Totally 129 patients with ILD and 143 coughing patients were included for the assessment. The ILD patients were all non-smokers and of connective tissue disease-associated interstitial lung disease（CTD-ILD）or interstitial pneumonia with autoimmune features（IPAF）. Lung function test results were compared between the two groups. Correlations of the diffusion capacity of the lung for carbon monoxide（DLCO）with other parameters were analyzed. Results：Restrictive and obstructive ventilation dysfunctions were observed in 51.2% and 2.3% of the patients with ILD，respectively. The vital capacity（VC），forced vital capacity（FVC），residual volume（RV）and total lung capacity（TLC）of the patients with ILD were significantly lower than those of the control group，but the forced expiratory volume in one second/forced vital capacity（FEV1/FVC）in the ILD patients was higher，which is of statistical significance（P < 0.05）. Residual volume/total lung capacity（RV/TLC）was similar in two groups（P > 0.05）. Maximal mid-expiratory flow（MMEF75/25）in the ILD group is higher but the difference is statistically insignificant（P > 0.05）. IOS airway resistance values are similar in two groups（P > 0.05）. DLCO is positively correlated with VC，FEV1，MMEF75/25，RV，TLC（P < 0.05），but negatively correlated with R5-R20，R5-R10（P < 0.05），and shows no correlation with FEV1/FVC. Conclusion：In non-smoking patients with CTD-ILD and IPAF，the restrictive ventilation dysfunction is overwhelmingly common compared with obstructive dysfunction. IOS is of no value in diagnosing ILD. With the progression of ILD and the declination of DLCO，the measured distal airway resistances using IOS show slight increase，but FEV1/FVC remain unchanged，suggesting air retention is unlikely and it is generally unnecessary to use bronchodilators during the course of CTD-ILD and IPAF.

Abstract:Objective：To evaluate the effects of different surface treatments on micro-structure and surface roughness of lithium disilicate glass-ceramic. Methods：After being sintered，routinely polished and washed，80 lithium disilicate glass-ceramic plates（12 mm×15 mm×2 mm）were randomly divided into three groups as follows：untreated，as control；sandblasted with 50 μm Al2O3；etched with 9.5% HF. The ceramic plates received different surface treatments were observed with a scanning electron microscope（sem）and their surface roughness were tested by a profilometer. Results：SEM showed that the surface of polished ceramics was relatively smooth，and after hydrofluoric acid treatment，the ceramic surface matrix dissolves，exposing two lithium silicate crystals and intersecting each other，alumina sandblasting after the ceramic surface could be seen irregular shallow concave structure；the measurement results of surface contour meter showed that the surface roughness of ceramics increased significantly after sand blasting and acid etching treatment（P < 0.05）. Conclusion：HF etching and sandblasting can increase the roughness of lithium disilicate glass-ceramic.

Abstract:Objective：To improve that traditional implants could hardly replace those nature-form teeth. Methods：By scanning standard model teeth，the three-dimensional finite element models of first molar teeth were established，and the vertical，horizontal and oblique three-dimensional instantaneous loads were applied to the model to obtain stress distribution and maximum stress of the tooth，mucosa，cortical bone and cancellous bone. Results：There is no significant difference of maximal stress between the first maxillary molar and first mandibular molar，but it is quite different from traditional cylindrical threaded implants. Conclusion：This result suggests that the implant with single form is difficult to meet the functional needs of different teeth，especially molar teeth，and future implant morphology can be expected similar to the natural tooth shape.

Abstract:Objective：To compare the difference of Hematoxylin eosin（HE） staining，P16 and Ki67 protein expression，as well as hTERC gene hybridization effect between environmental transparent dewaxing agent Van-clear and traditional transparent dewaxing agent xylene in different cervical lesions，and to explore the practical value of Van-clear. Methods：A total of 42 cervicitis specimens，53 cervical intraepithelial neoplasia（CIN）Ⅰspecimens，61 CIN Ⅱ specimens，56 CIN Ⅲ specimens，and 48 cervical cancer specimens were collected from Shantou Central Hospital from January 2016 to July 2018. In group A，260 paraffin sections were made from xylene，while in group B，260 paraffin sections were made from Van-clear. HE staining，immunohistochemistry（IHC）staining，and fluorescence in situ hybridization（FISH）of hTERC gene were used to detect the difference of HE staining excellence rate，P16 and Ki67 protein expression，and hTERC gene positive rate between the two groups. Results：①Sections in group A and group B for HE staining and immunohistochemistry staining all met the quality control standard of industry. ②The excellent and good rates of HE staining in group A and group B were 98.08% and 99.23%，respectively. There was no significant difference between the two groups（χ2=1.30，P=0.25）. ③Comparison of positive rate of IHC staining showed there was no significant difference in the positive rate of P16 and Ki67 protein in different cervical lesions between groups A and gronp B（P＞0.05）.④Comparison of the positive rate of hTERC gene amplification showed there was no significant difference between group A and gronp B in different cervical lesions（P＞0.05），and no significant difference was observed between the two groups under fluorescence microscope. Conclusion：Van-clear，as a new environmental transparent dewaxing agent，has the same effect as traditional transparent dewaxing agent xylene in HE staining，P16 and Ki67 protein expression，and hTERC gene detection in different cervical lesions. It is also green and environment-friendly，we recommend that it be widely used in clinic.

Abstract:Objective：This study aims to establish an immunological method for detecting specific antibody to Aspergillus fumigatus thioredoxin reductase（TR）in human sera，and evaluate its value in clinical prospect. Methods：The double-antigen sandwich ELISA for detecting TR antibody was applied with recombinant TR as coating and enzyme-labelled antigen. Reaction conditions were optimized by chessboard titration，and the sensitivity，specificity and repeatability for the ELISA were determined. Sera from 273 inpatients（50 cases with invasive aspergillosis，46 cases with invasive candidiasis，82 cases with Candida colonization and 95 cases with bacterial infection）and 200 healthy volunteers were collected and tested by the sandwich ELISA. The experiments results were compared with the previous results of indirect ELISA. Results：The TR was labeled with horseradish peroxidase（HRP）by the reformative sodium periodate method and the optimized condition were as follows：the coating concentration of TR was 1.0 μg/mL，the dilution of enzyme-labelled antigen was 1∶500，the blocking buffer was PBS-T containing 50 g/L dried skimmed milk，test serum was diluted to 1∶100. The intra- and inter-assay coefficients of variation for the developed ELISA were 11.2%，11.8%，12.3% and 10.9%，12.1%，13.5%，respectively. The blocking rate of sera with recombinant antigen was 94.4%. The absorbance value of 0.126 was chosen as cut-off value according to the ROC curve. The sensitivity of the sandwich ELISA for diagnosis of invasive aspergillosis was 80.0%（40/50），higher than that of indirect ELISA（83.3% vs. 72.2%） and the specificity was 95.5%（404/423） in this condition. Conclusion：A double-antigen sandwich ELISA for detecting the specific antibody to Aspergillus fumigatus TR was established successfully，which can be potentially applied in clinical diagnosis setting of invasive aspergillosis.

Abstract:Near-infrared fluorescent materials include traditional molecular dyes and new quantum dots. Near-infrared spectroscopy belongs to the frequency doubling and main frequency absorption spectroscopy of molecular vibration spectroscopy. It is mainly produced by the non-resonance of molecular vibration when molecular vibration transits from ground state to a higher energy level，and possesses strong tissue penetrability. In recent years，near-infrared fluorescent materials modified by molecular biological techniques have become a research hotspot in medical imaging diagnosis. They have strong tissue penetrability，low background fluorescence interference，high fluorescence signal sensitivity，spectral stability，quenching resistance，molecular targeting recognition and good biological safety. In this paper，the construction of near-infrared fluorescent material and its application in clinical imaging examinations such as ultrasonography，CT，MRI and PET are analyzed. The diagnostic advantages and key techniques of multimodal contrast strategy are discussed，which provide theoretical basis for further clinical application.

Abstract:Interleukin（interleukin，IL）-33 can be expressed in various tissues and cells. It plays an important role in the maintenance of homeostasis and the response to environmental stress. It can participate in the formation and development of many diseases through IL-33/ST2 signaling pathway. Studies have shown that IL-33 is involved in maternal and fetal interface immune tolerance and placenta formation，while abnormal expression of IL-33 is related to the formation and development of pregnancy-related diseases such as abortion，preterm delivery and preeclampsia. The roles of IL-33 in pregnancy are deeply studied，and it may provide new ideas for the diagnosis and treatment of normal pregnancy maintenance and pregnancy specific diseases.

Abstract:Long non-coding RNA（lncRNA）has become a research hotspot in recent years，and it has been proved that abnormal expressions of some certain lncRNAs are closely related to the occurrence and progression of tumors. Among many kinds of lncRNAs，the metastasis associated lung adenocarcinoma transcript 1（MALAT1）is upregulated in metastatic carcinoma cells. MALAT1 was widely expressed in normal tissues，and the expressions displayed significant differences in a variety of human tumor tissues，cells and peripheral blood，which suggests that MALAT1 may play an important role in the process of tumorigenesis，progression，invasion and metastasis. Herein we document the molecular characteristics and functions of MALAT1 with a reference to their implications in the molecular pathology of various cancers.