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Objective@#To investigate the effects of Tim-3 on osteoclast-like cell (OLC) formation and bone resorption induced by peripheral blood mononuclear cells (PBMCs). @*Methods@#The expression levels of Tim-3 in of rheumatoid arthritis (RA) patients and healthy controls were detected by flow cytometry. The OLCs were induced by human PBMCs in vitro. The expression levels of tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTSK) and matrix metalloproteinase 9 (MMP9) mRNAs in the formation of OLCs were detected by real-time quantitative PCR. The morphology of OLCs was observed by Wright′s staining and G-actin staining, and the number of OLCs was counted by TRAP staining. The number and area of bone resorption pits in OLCs were detected by the Corning Osteo Assay Surface. @*Results@#The expression levels of Tim-3 in PBMCs of RA patients ([77.31±10.66]%) were significantly higher than that of healthy controls ([51.72±16.69]%, t=7.593, P<0.01). When PBMCs with different Tim-3 levels were induced into OLCs, the area of bone resorption pits in the high Tim-3 level group ([1.054±0.085] S/mm 2 ) were significantly lower than those in the intermediate Tim-3 level group ([1.889±0.053] S/mm 2 ) and the low Tim-3 level group ([2.763±0.066] S/mm 2 , F=9.318, P<0.05). @*Conclusion@#Tim-3 may negatively regulate the bone resorption of OLCs.
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Objective To observe and analyze the expression of programmed cell death 4 (PDCD4) gene and apoptosis inhibitor Livin in triple negative breast cancer (TNBC) tissues and its relationship with prognosis.Methods One hundred cases of TNBC tumor tissue,50 cases of adjacent carcinoma tissue,50 cases of normal breast tissue were selected as the research data.The immunohistochemical technique was applied to detect and compare the expression positive rates of PDCD4 and Livin protein in three kinds of tissues.The patients were followed up.The overall survival (OS) and the progression free survival (PFS) were observed and compared.Results The expression positive rate of PDCD4 in TNBC tissue was significantly lower than that in adjacent carcinoma tissue or normal breast tissue,the differences were statistically significant (x2=26.613,32.000,P<0.05).The expression was correlated with the clinical pathological features of tumor size,lymph node metastasis,clinical stage,axillary lymph node metastasis and cancer embolus (x2=26.936,13.210,22.774,27.463,5.803,P<0.05);the expression positive rate of Livin protein in TNBC tissue was significantly higher than that in adjacent carcinoma tissue or normal breast tissue and the expression positive rate of Livin protein in adjacent carcinoma tissue was significantly higher than that in normal breast tissue,the differences were statistically significant (x2 =14.614,57.353,19.048,P<0.05).The expression was correlated with the clinical pathological features of lymph node metastasis,clinical stage,axillary lymph node metastasis and cancer embolus (x2 =10.788,6.160,27.350,8.914,P<0.05);OS,PFS in the patients with PDCD4 negative expression were significantly lower than those in the patients with PDCD4positive expression.OS,PFS in the patients with Livin positive expression were significantly lower than those in the patients with Livin negative expression,the above differences were statistically significant (x2 =23.931,19.163,22.649,17.213,P<0.05).OS in the TNBC patients was correlated with age (RR=1.405),clinical stage (RR =2.897),tumor diameter (RR=2.722),axillary lymph node metastasis (RR=2.516),vascular invasion (RR=3.020),PDCD4 Expression (RR=1.752) and Livin expression (RR=2.051) (P<0.05).PFS in the patients was correlated with clinical stage (RR =2.756),axillary lymph node metastasis (RR =2.437),PDCD4 expression (RR =1.649) and Livin expression (RR=1.804) (P<0.05).Conclusion The PDCD4 low expression and Livin protein over-expression exist in TNBC tissues.Their abnormal expressions are correlated with the clinicopathological features of tumor and the prognosis of patient,and could be used as the auxiliary indexes in evaluation of progression and prognosis of TNBC.
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Objective To investigate the molecular characteristic of fluoroquinolone resistant Streptococcus agalactiae (GBS) isolates in Suzhou.Methods Totally 46 fluoroquinolone resistant GBS strains were collected,and then subjected into PCR and two resistance genes were sequenced and their high frequency mutation sites,gyrA and parC were analysed.Results Among these isolates,the most frequent gyrA mutation was gyrA_S81L (TCA→TTA),which displayed 93.4% (43/46) with substituted at No.81 site of amino acid from Ser to Leu.However,the most frequent parC mutation was mutated at No.79 site of amino acid from Ser to Tyr orPhe,with two mutations parC_S79Y (73.9%,34/46) and parC_S79F(17.4%,8/46).The analysis showed that the major mutation patterns are gyrA_S81L with parC_S79Y (73.9%,34/46),gyrA_S81L with parC(13.0%,6/46).Conclusion The most frequent mutations of fluoroquinolone resistant gene in Suzhou are gyrA and parC and the major mutation patterns are gyrA_S81L/parC_S79Y and gyrA_S81L/parC_S79F.
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Objective To observe the repeat administration toxicity of Guanxinning Injection in rats.Methods Totally 120 rats (males and females) were randomly divided into vehicle control group and three Guanxinning Injection groups with different dosages (3.40,1.70,and 0.85 g/kg).Rats were administered with Guanxinning Injection by consecutive intravenous injection for 13 weeks.Besides the general conditions were observed,the related indexes were detected,such as body weight,the routine control of blood,hepatic function,renal function,the metabolism condition of lipids,the glycometabolism indexes,and histopathology analysis were determined at 13 weeks of treatments and 4 weeks after the withdrawal,respectively.Results Rats in the Guanxinning Injection group at 3.40,1.70 g/kg apperance shortness of breath,unsteady gait,lying motionless,and other symptoms.There was no obvious abnormal reaction in the 0.85 g/kg dose group.There were two male rats in the Guanxinning Injection group at 3.40 g/kg died at about week 4 of treatments,and there was no death in the 0.85 and 1.70 g/kg dose groups.Compared with vehicle control group,the related indexes of blood,blood biochemistry,organ relative quality,and histopathological showed no obvious abnormalities in Guanxinning Injection 0.85 and 1.70 g/kg group.The levels of urea nitrogen (BUN) and the relative weight of kidney in Guanxinning Injection 3.40 g/kg group were significantly higher than those in vehicle control group at 13 weeks.The change of these parameters regained to normal at 4 weeks after withdrawal and the rest of the detection index showed no obvious abnormality in 3.40 g/kg dase group.Conclusion Intravenous administration of Guanxinning Injection for 13 weeks at high dose could induce reversible damage to kidney.
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Objective To explore the differences and similarities of the cervical lesions and mechanism between Asian variant E6 T178G and European variant E6 T350G, A442C and other variants. Methods We selected 300 clinic or hospitalized patients in our hospital during the period of May 2011 to October 2012. Cervical exfoliated cells were harvested by Thinprep cytologic test (TCT). A PCR sequencing assay was performed to detect HPV16 E2, E6 and E7 gene variants. One year later, the test was repeated. The patients with persistent infection underwent cervical biopsy by colposcopy for pathological examination. SP immunohistochemical method was applied to detect E7 protein expression level in all the patients. Results After one year, of 292 patients who were successfully sequenced, 259 were chronic cervicitis, 32 were cervical intraepithelial neoplasia grade I (CINI), and one was cervical intraepithelial neoplasia grade II (CINII). E7 protein expressed in each variant. But the expression of E7 protein in patients with different variant infection had no significant difference from each other. Conclusions E7 protein may be play a role in the early stages of HPV16?induced cervical lesions. But E7 protein may not be a reference index of the different carcinogenic mechanism between different HPV16 variants.
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Objective To investigate the synergism and attenuation effects of Pini Pollen on cyclophosphamide chemotherapy in model mice.Methods The models of mouse were induced with S180 tumor and cyclophosphamide.The tumor inhibiting rates, hematology, Liver function, giant cell phagocytosis, hemolysin level and indexes of thymus and spleen were determined to evaluate the synergism and attenuation effects of Pini Pollen(150,300,600 mg/kg) on cyclophosphamide chemotherapy.Results Compared with cyclophosphamide chemotherapy group,the tumor inhibiting rates,white blood cell number,giant cell phagocytosis, hemolysin level and indexes of thymus and spleen were increased, at the same time liver function damage were alleviated significantly in cyclophosphamide plus Pini Pollen groups.Conclusions Pini Pollen has a remarkable synergism and attenuation effects on cyclophosphamide chemotherapy through its immune adjustment functions.
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Objective to investigate the role of Pim‐3 and NF‐κB in the development and progression of infiltrating ductal carcinoma of the breast .Methods Here ,we used immunohistochemistry to detect expression of Pim‐3 and NF‐κB in 75 samples of infiltrating ductal breast carcinoma ,21 samples of intraductal breast carcinoma and 30 normal breast tissues .The relationship of their expression ,as well as their correlation with clinicopathological features and patient survival were assessed .Results In con‐trast ,both Pim‐3 and NF‐κB were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue .In the infiltrating ductal carcinoma ,the positive expression rate of Pim‐3 was 77 .3% ,and that of NF‐κB was 68 .0% ;in duc‐tal carcinoma of the breast ,the positive expression rate of Pim‐3 was 52 .4% ,and that of NF‐κB was 42 .9% ;in the normal breast tissue ,the positive expression rate of Pim‐3 was 23 .3% ,and that of NF‐κB was 16 .7% ;the positive expression rate of Pim‐3 was correlated with tumor size ,histological grade ,and clinicopathological stage ;and that of NF‐κB was correlated with tumor size ,histo‐logical grade ,lymph node metastasis of breast cancer .Spearman rank correlation analysis revealed a positive correlation between Pim‐3 expression and NF‐κB expression in infiltrating breast cancer (r=0 .243) .Conclusion Our results demonstrate that Pim‐3 and NF‐κB play a role in the initiation and development of breast cancer ,thus ,these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer .
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Objective To investigate the repeated dose toxicity of doxorubicin liposome injection and doxorubicin injection in rats. Methods Ninety SD rats ( body weight 180-220 g, male:female=1:1 ) were divided into 3 groups (30 rats in each group), and were administered intravenously with physiological saline, doxorubicin liposome injection (1 mg·kg-1 ) and doxorubicin injection ( 1 mg·kg-1 ) , respectively, once every three days for thirteen times. The body weight, blood biochemistry, hematology, organ coefficient and histopathology were analyzed for the overall toxicity assessment. Results The rats administered with doxorubicin liposome injection (1 mg/kg) showed hair loss and skin ulcer, significantly reduced growth of body weight, increased levels of urea nitrogen ( BUN ) , alanine aminotransferase ( ALT ) , blood platelet ( PLT ) , and kidney and heart coefficients, decreased thymus and testicular coefficients, myofibrillar rupture and lysis, and partial loss of cell nuclei, hyaline casts in the renal convoluted tubules, interstitial edema and loss of spermatogenic cells in the testicular tubules. Compared with the doxorubicin liposome injection group, similar abnormal changes were also observed in the doxorubicin injection group, but the hair loss and skin ulcer were milder and the heart and kidney toxicities were severer. Conclusions Compared with doxorubicin injection, the doxorubicin liposome injection causes milder heart and kidney toxicity but more serious skin toxicity.
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Objective To investigate the therapeutic mechanism of umbilical cord mesenchymal stem cell(UC-MSCs)trans-plantation for the graves disease(GD)mice .Methods Thirty two mice were divided into 3 groups as following :normal control group (G0) ,GD control group (G1) ,UC-MSCs group(G2) .Enzyme linked immunosorbent assay(ELISA)was used to measure the level of TSAb in blood serum and the expression of FT4 was measured by chemiluminescence .Thyroid sections were stained with hema-toxylin and eosin(HE)for histological examination .Splenocytes were stained with multicolor immunofluorescence and detected by flow cytometry to analyze the percentages of CD1dhiCD5+CD19+ regulatory B cells(Bregs) .Expressions of IL-10 and TGF-βmR-NA in spleen organization were measured by Real-time PCR .Results At 26 weeks ,the level of TSAb in blood serum in G2 was more significantly decreased than in G1(P<0 .05) ,and the level of CD19+ B in spleen in G2 was also more significantly decreased than in G1(P<0 .05) ,however ,the percentage of CD1dhiCD5+CD19+ Bregs splenocytes and the levels of IL-10 and TGF-βmRNA in spleen organization were more significantly increased than in G1(P<0 .05) .The concentration differences of TSAb in serum was negatively correlated with the percentage differences of CD1dhi CD5+ CD19+ Bregs ,however ,positively correlated with the expres-sion differences of IL-10 and TGF-βmRNA in spleen before and after transplantation .Conclusion Activation of Bregs may be one of the mechanisms of UC-MSCs therapeutic effect on GD mice .
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Objective To investigate the in vivo effects of artesunate (ART) treatment on OAZ gene expressions in MRL/lpr lupus mice.Methods Twenty-four 12-week-old female MRL/lpr lupus mice were randomly divided into two groups by the random number table,i.e.,the lupus control group and the ART treatment group,12 in each group.Besides,another 8 BABL/C mice were recruited as the normal control group.Peripheral blood mononuclear cells (PBMC) were collected from lupus mice before treatment as well as 4 week and 8 week after treatment,and RNA was extracted and reverse transcripted to cDNA.mRNA expression levels of OAZ and Id1-3 were measured by using real-time polymerase chain reaction (PCR),and the data between the two groups were analyzed by t test,a plurality of samples were compared with the single factor analysis of variance.Serum levels of IFN-γ,IL-4,BAFF and ANA were tested by enzyme-linked immunosorbent assay (ELISA).Relationships of the gene expression levels with levels of cytokines and ANA were analyzed.Statistical analysis were uising t test,ANOVA and LSD-t test.Results mRNA expression levels of OAZ,Id1 and Id3 gene (△Ct:12.9±0.8,12.0±0.4,10.2±0.8) in lupus mice were significantly different from 8 weeks after the treatment comparing to those in the lupus control group (△Ct:9.8±1.0,9.3± 1.1,8.1±0.8) and the normal control group (△Ct:13.9±1.2,11.4±0.7,4.7±0.8,10.3±1.0)(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),and alteration of OAZ mRNA expression levels before and after the treatment were positively correlated with changes of Id1,Id3 levels (r=0.867,0.947; P<0.05),and levels of IL-4 and ANA were significantly lower 8 weeks after the treatment than those in the lupus control group (P<0.05); while level of IFN-γwas higher than those in the lupus control group (P<0.05).Alteration of OAZ mRNA expression levels before and after the treatment were negatively correlated with changes of ANA,BAFF levels (r=-0.955,r=-0.937; P<0.05) and positively correlated with changes of Th1/Th2 levels (r=0.976,P<0.01).Conclusion The expression of genes involving in the OAZ and downstream gene are effectively reduced along with the alteration of several cytokines and ANA after ART treatment.OAZ signaling pathway can play an important role in ART treatment for SLE.
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With the understanding of the development and pro-gress of the cancer,the research of targeted cancer drug devel-opment reaches into a new era.p53 is an important tumor sup-pressor gene,the protein coded by p53 plays a critical role in tumor suppression mainly by inducing cell cycle regulation, DNA repair and apoptosis.Nowadays,p53 becomes a relatively attractive target for anti-cancer drug development and there are some drugs targeting p53,moreover,APR-246 which targets mutant p53 is in Phase II clinical trial.In addition,it facilitates drugs discovery programmes in the challenging area of protein-protein interactions and mutant protein conformational change. The review discusses the research progress of drugs which target p53 and elucidates the characteristics and mechanisms of these compounds.
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In the reform of human resource system at public institutions,public hospitals are challenged with changing employment mechanism and effective mobilization of all-staff's incentives.This paper introduced the classified staff management by Sir Run Run Shaw Hospital,College of Medicine,Zhejiang University,which covered the background,specific methods,purposes,initial results,as well as the key links and problems encountered.The study proved that classified management of the hospital staff helps create a fair and impartial workplace,conducive for sustainable development of the hospital.
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Study of the strategic objectives chosen by hospitals from the perspective of stakeholders is now under the spotlight of researchers at large. To identify whether there exist differences in strategic objectives among various groups in the hospital and probe into the causes, the author called into play the competing values framework as the analysis tool. An evidenced-based study on public hospitals in Hangzhou identified marked differences on choices of strategic objectives among various internal stakeholders in the hospital For example, hospital administrators tend to focus on internal control, while medical doctors tend to focus on interactions between the hospital and the external environment. On this basis, further studies are proposed.
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Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.
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Adoption of the board of directors system in a public hospital is conducive to straightening out its internal management system and enables its head to more effectively exercise his power of supervision while giving full play to his functions. In addition, when major issues are concerned, decision-making is more democratic and scientific, reducing disputes over trifles. The board of directors system may after all be accepted as an effective form of management.