Objective To investigate the regulation of gene variation and explore the tumor associated suppressor genes on chromosome 9 during the developing of cervical squamous cell carcinoma. Methods Microdissection, PCR, electrophoresis and radiograph were selected to detect the LOH in 46 cases cervical squamous cell carcinoma with high-grade squamous dysplasia and normal tissue. The changes of LOH at chromosome 9 total seven microsatellite markers and relationship between LOH rate and clinical parameters were analysed. Results Total frequency of LOH in CIN (Ⅱ,Ⅲ) was 16 % and in cervical squamous cell carcinoma was 25 %. In the former, LOH at marker D9S171(30 %), D9S162(23 %), D9S43(20 %), D9S303(17 %), D9S753(12 %), D9S242(11 %) were found, whereas D9S1748 LOH was not detected in high-grade dysplasia. In the latter, LOH at marker D9S171(41 %), D9S43(33 %), D9S162(31 %), D9S242(24 %), D9S303(17 %), D9S753(17 %), D9S1748 13 % were found. In addition, LOH was found in high-grade dysplasia in three cases but not in cervical squamous cell carcinoma. Conclusions The study in seven microsatellite markers showed that from normal squamous tissue to dysplasia to cervical squamous cell carcinoma were accompanied with accumulation of gene errors. The p15 gene inactivation had a high relationship with the occur of cervical squamous cell carcinoma. Tumor suppressor genes associated with cervical squamous cell carcinoma may exist near or at D9S43, D9S162, D9S242. Some cases showed that high-grade dysplasia and cervical squamous cell carcinoma may came from different independent clone which provide some clue for multiple foci carcinoma at genetic level.

Objective: Dithiothreitol(DTT) and ?-Mercaptoethanol(2-ME) are important reducing agents for SDS-PAGE.This study is to observe the diffusing effect of DTT and 2-ME during electrophoresis,and to find a way of avoiding this effect.Methods: We placed protein samples containing reducing agents and non-reduced protein samples separately in the sample wells at intervals for SDS-PAGE electrophoresis,and determined whether or not the electrophoretic lanes of the non-reduced samples were interfered by the adjacent lanes.Results: DTT and 2-ME diffused to the neighboring lane,so that the non-reduced samples were reduced partially.The spreading effect was positively correlated with the content of the reducing agent.Conclusion: DTT and 2-ME have a diffusing effect during SDS-PAGE electrophoresis.In separating the reduced and non-reduced proteins in the same gel at the same time,at least a blank lane should be set up in between them in order to avoid the diffusing effect.

Objective To investigate the pathogens distribution in neonatal blood culture and their drug resistance to antibacteri‐al drugs in Dalian City .Methods The routine blood culture ,identification and drug sensitivity test were carried out in the hospital‐ized neonates from August 2014 to August 2015 .And the obtained data were analyzed .Results A total of 186 strains of pathogenic bacteria were detected from 1 570 cases of neonatal blood culture and the positive rate was 11 .8% .Gram positive bacteria accounted for 74 .2% (138/186) and were dominated by Staphylococcus epidermidis .Gram negative bacteria accounted for 25 .3% (47/186) , which were mainly onion burkholderia bacterium .One strain was fungus ,accounting for 0 .5% .The drug sensitivity test results showed that Gram positive bacteria had the higher resistance rate to penicillin and erythromycin (80 .0% -90 .0% ) ,100 .0% sensi‐tivity to vancomycin ,linezolid and teicoplanin ;E .coli and K .pneumoniae had the highest resistance rate to ampicillin (88 .2% -100 .0% ) ,100 .0% sensitivity to imipenem ,amikacin ,low resistance rate to piperacillin/tazobactam ,cefoperazone/sulbactam ,amoxi‐cillin/clavulanic acid ,cefepime and ceftazidime (0% -10 .0% ) .The resistance rate of onion burkholderia bacterium to ticarcillin/clavulanate and meropenem was higher than 80 .0% ,which had 100 .0% sensitivity to cefoperazone /sulbactam ,levofloxacin ,mino‐cycline and compound sulfamethoxazole .Conclusion The neonatal blood culture pathogen in Dalian City is dominated by Gram pos‐itive bacteria ,coagulase negative staphylococcus is the main pathogen .Due to the different regional environmental ,pathogens and drug resistance should be regularly monitored and analyzed to provide objective and accurate basis for clinical rational use of anti‐bacterial drugs .

Aim To study the influence of Sodium fer-ulate ( SF) on bone metabolism in glucocorticoid–in-duced osteoporosis rats. Methods Thirty cases of fe-male Wistar Rats(3-month-old) were divided into con-trol group, model group and SF group ( low-dose group, middle-dose group, high-dose group ) by ran-domized block design. Double fluorochrome labeling with calcein was performed before necropsy. The left tibia was taken for bone histomorphometry. Results In static parameters, the proximal tibia cancellous bone trabecular thickness, trabecular quantity and area ratio were significantly reduced in model group compared with control group;while compared with model group, those were increased in middle and high-dose SF group. Trabecular separation degree was increased in model group compared with control group, while it was decreased in middle and high-dose SF group compared with model group. In dynamic parameters, the calcula-tion parameters of cancellous bone mark perimeter rate and the bone formation rate were increased in model group compared with control group, in middle and high-dose SF group the bone formation rate was in-creased compared with model group. In bone cells, os-teoclast number per mm, osteoblast number per mm, percent osteoblast surface perimeter and percent osteo-clast surface perimeter were increased in model group compared with control group. In growth-plate, the thickness of growth-plate was increased in model group compared with control group. In bone cells and growth-plate there was no statistical significance between treat-ment group and model group. Conclusion This study demonstrates that SF can increase bone mass and im-prove bone structure,which may be related to the im-provement of bone formation. SF is effective for GIOP in rats.

Objective To investigate the expression of suppressor of cytokine signaling genes (SOCSs) and JAKs mRNA in the acute myloid leukemia(AML) patients. Methods The expression of SOCSs and JAKs mRNA as well as TYK2 in AML patients and healthy adults as normal contrals (NC) was measured with RT-PCR. Results The expression of SOCS 1,4,5 and 7 in AML patients was lower than those in normal control and AML with remis-sion (P<0.01),but the expression of SOCS 3 and 6 was higher than those in normal control and remission AML(P<0.01),however there was no significant difference in SOC2 between groups. The expressions of JAK2 ,JAK3 and TYK2 in AML were significantly higher than those in patients with remission and normal control(P<0.05). The ex-pression of JAK1 mRNA in relapsed AML was higher than that in normal control group(P<0.05),but the latter has no statistical significance between beginning treatment and normal group(P>0.05). Conclusion The deletion and degradion of SOCS 1,4,5 and 7 present in AML patients and JAKs expression is significantly increased, suggesting that both of them may co-participate in the pathogenesis of AML.

Obej ctive Abnormal proliferation of T cells plays an important role in the development of autoimmune diseases. The article aimed to study the inhibitory effect of small molecule compound BD691 on T cell proliferation and its mechanism. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads,then T cells were ac-tivated with anti-CD3/CD28 mAbs or alloantigen.The inhibitory effect of BD691 on activated T cell proliferation, the cytotoxic effect BD891 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometry.Furthermore, ELISA was used to detect the secretion of cytokines associated with T cell differentiation. Results BD691 significantly inhibited the prolif-eration of T cells being stimulated by anti-CD3/CD28 mAb or alloantigen in a dose-dependent manner, and IC50 values are (8.5 ± 1.5)μmol/L and (7.2 ±1.3)μmol/L, respectively.However, BD691 had no obvious cytotoxic effects on resting T cells and periph-eral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .In T cells which were not activated by anti-CD3/CD28 mAb, the percentage of CD25+T cells is only 1.6%of the total cells, while the number increased to 68% after activating treatment.Mean-while, in T cells which were activated by 0, 3.3, 10, 30μmol/L BD691, no obvious change of CD25 expression were observed, while immunosuppressant FK506 (0.1μmol/L) significantly decreased the expression of CD25 +T cells (14.9%).In unactivated T cells, 95.6%cells were at G0/G1 phase, while after activation, the percentage of cells at G0/G1 phase reduced to 57.7%.In addition, BD691 inhibited the secretion of IFN-γ, IL-6 and IL-17 in activated T cells, but had no effects on the secretion of IL-2, IL-4 and IL-10. Co nclusion BD691 exerts no effects on T cell activation, but it inhibits T cell proliferation by inducing T cell cycling arrest at G0/G1 phase.Moreover, BD691 inhibits the secretion of key cytokines (such as IFN-γ, IL-6, IL-17) closely related to the differ-entiation of Th1 and Th17 cells.The results suggest that BD 691 is a potential lead compound to develop a new immunosuppressant for the inhibition of abnormal proliferation and differentiation of T cells.

Objective Benzothiazole derivative BD960 has immunosuppressive activity after cell -based assays for high-throughput screening.The paper aimed to investigate the involved mechanism of BD960 on T cell proliferation. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads.Then the T cells were activated by anti-CD3/anti-CD28 mAbs or alloantigen.The effect of BD960 on activa-ted T cell proliferation, the cytotoxic effect BD960 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometer.Cytokine levels, including IL-2, IL-4, IL-6, IL-10, IL-17A and IFN-γ, were determined by ELISA. Results BD960 significantly inhibited the proliferation of T cells stimulated by anti-CD3/anti-CD28 mAb or alloantigen in a dose-dependent manner.The IC50 value is (2.3 ±0.3)μmol/L or (2.5 ±0.3)μmol/L, respectively.Moreover, BD960 had no obvious cytotoxic effects on rest-ing T cells and peripheral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .The ratio of CD25 expression on T cell was 69.7%after stimulated by Anti-CD3/CD28 mAbs with 72 h, the concentration (0.625、2.5、10)μmol/L of BD960 also had no potent effects on the ratio, but 0.1μmol/L FK506 could inhibit CD25 expression as low as 9.4%.The G0/G1 phase of activated T cells was 58.5%after stimulated by BD960 with 96 h.BD960 could induce cell cycle arrest at the G0/G1 phase in activated T cells with the increase of concentration and RAPA in the concentration of 0.1 μmol/L was 91.5%.In addition, BD960 (0.625、2.5、10)μmol/L could inhibit the secretion of IFN-γ, IL-6 and IL-17 in activated T cells with the increase of concentration, without any effects on the secretion of IL-2, IL-4 and IL-10. Conclusion BD960 not only exerts the inhibition on the late stage of T cell activation of cell proliferation but also inhibits the secretion of inflammatory cytokines, such as IL-6, IL-17 and IFN-γ, while the mechanism of BD960 on T cell proliferation was not the same as FK506.As a result, BD960 has the potential to be the lead compound to develop a new immunosuppressant.

Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

Objective To observe the effect and adverse effect of ultrashort wave combined with Etroricox-ib in treatment of periarthritis of shoulder. Methods 80 cases with periarthritis of shoulder from March 2015 to March 2016 were randomly assigned into treatment group and control group. Etroricoxib therapy was provided in control group while ultrashort wave plus Etroricoxib therapy in treatment group. VAS ,ROM and MBI were applied for the evaluation before and after the treatment. The differences of clinical cure rate ,recurrence rate and adverse reactions were observed between 2 groups. Results There were higher cure rates ,lower recurrence rates and less adverse reactions in 2 groups after the treatment. The shoulder joint pain ,rang of should motion and BI of both groups were improved with significant differences after the treatment ,but treatment group witnessed more improve-ment(P<0.05). Conclusions Ultrashort wave combined with Etroricoxib therapy can relieve shoulder pain ,and further improve the function of shoulder joint activity in the treatment of periarthritis of shoulder. It is recommended for clinical application.

Objective? To explore the construction method and the effects of the orthopedic specialized nurse studio. Methods? The orthopedic specialized nurse studio included 4 working groups, which were intravenous therapy group, wound care group, chronic diseases management group and perioperative venous thromboembolism (VTE) management group. Each group were assigned 1 team leader and 10 to 12 team members. Nursing training, management and research were carried out. The professional competency of orthopedic nursing staff and the nursing quality were compared before (January to December 2016) and after (January to December 2017) the construction of the studio. Results? The scores of theoretical assessment, specialized nurses' performance and nursing quality in 2017 were (97.76±4.09), (82.25±6.60) and (99.06±0.91) respectively, which were higher than those of the same period in 2016. The differences were statistically significant (t=2.169, 3.327, 2.129; P＜ 0.05). Conclusions? The establish of the orthopedic specialized nurse studio can enhance the growth of specialized nursing team, improve the nursing quality and promote the development of multi-disciplinary integration.