Objective To investigate the expression and clinical significance of p16 protein(p16) ,Ki-67 antigen(Ki-67) and cytokeratin 7(CK7) in cervical precancerous lesion .Methods The paraffin embedded tissue samples in 96 cases of cervical intraepithelial neoplasia(CIN) from January 2014 to March 2017 were selected and divided into the CIN Ⅰgroup(25 cases) ,CIN Ⅱgroup (36 cases) and CIN Ⅲ group(35 cases) according to the lesion severity ;meanwhile the paraffin embedded tissue samples in 64 ca-ses of chronic cervicitis were selected as the control group .The positive expression rates and expression modes of p16 ,Ki-67 and CK7 in different degrees of cervical lesion tissue were detected by adopting the immunohistochemistry method .Results The positive expression rates of p16 ,Ki-67 and CK7 in the CINⅠ ,Ⅱand Ⅲgroups were significantly higher than those in the control group , the difference was statistically significant (P＜0 .05) .The total score of p16 ,Ki-67 and CK7 had consistency with the degree of cervical precancerous lesions(Kappavalues were 0 .546 ,0 .628 and 0 .349 respectively) .The receiver operating characteristic curve analysis results showed that the area under the curve of p16 ,Ki-67 and CK7 combined detection was higher than that of single detection of these 3 indexes(0 .978 vs .0 .930 ,0 .966 ,0 .837) ,the difference was statistically significant (P＜0 .05) .Conclusion The expression of p16 ,Ki-67 and CK7 has the correlation with the degree of cervical precancerous lesions .Their joint detection is helpful to improve the diagnostic efficiency of cervical precancerous lesions and could be used in the assisted diagnosis of cervical lesion and prediction of high risk progress cervical precancerous lesion .

Objective To investigate the effect of bronchofibroscopic bronchoalveolar lavage on serum inflammatory cytokines in the patients with severe ventilator-associated pneumonia(VAP) .Methods 68 cases of severe VAP were divided into the control group and observation group according to the random number table method ,34 cases in each group .The control group used conventional alveolar lavage plus sputum suction treatment ,while the observation group adopted bronchofibroscopic bronchoalveolar lavage plus sputum suction treatment .The lung infection control time ,mechanical ventilation treatment time ,time of body temperature recovery to normal ,pulmonary infection score(CPIS) ,white blood cell(WBC) count ,respiratory function ,blood gas indexes and serum inflammatory factors levels were compared between the two groups .Results The lung infection control time ,mechanical ventilation treatment time ,recovery time of normal temperature ,blood gas indexes and serum inflammatory factors levels after treatment in the observation group were significantly superior to those in the control group (P＜0 .05) .Conclusion Bronchofibroscopic bronchoalveolar lavage can significantly improve the respiratory function and blood gas indexes ,reduces the serum inflammatory factors levels in the patients with severe VAP .

Objective To reveal the pancreatic cancer cell drug resistance mechanism to provide a basis for clinical treatment by studying miR-26b targeting p53 for promoting the drug resistance of pancreatic cancer cell line PANC-1 .Methods (1) The over-expression and knockdown plasmid of p53 and the fluorescent reporter vector containing 3′Untranslated region(3′UTR) were constructed respectively .(2) The effect of p53 and miR-26b on the growth and proliferation of PANC-1 was investigated by methyl thiazolyl tetrazolium(MTT) in the presence of gemcitabine .(3) The target relationship between miR-26b and p53 was determined by bioinformatics ,real-time polymerase chain reaction(PCR) ,fluorescent reporter vector and Western blot experiment .(4) The effect of p53 on the growth of miR-26b was investigated by salvage experiments .Results (1) The MTT experiment confirmed that ,in the presence of gemcitabine ,over-expression of p53 could inhibit the proliferation of pancreatic cancer cell line PANC-1 ,and knock-down of p53 could promote the growth and proliferation of PANC-1;(2) the bioinformatics prediction showed that miR-26b targeted p53 ,real-time PCR ,Western blot and fluorescent reporter vector experiment confirmed that p 53 is the target gene of miR-26b , and miR-26b inhibits transcription and translation of p53 by targeting the 3′UTR of p53 gene;(3) the MTT experiment confirmed that in the presence of gemcitabine ,over-expression of miR-26b could promote the growth and proliferation of PANC-1 cells ,and enhanced its resistance to gemcitabine ;(4) the rescue experiment confirmed that the simultaneous over-expression of p53 rescued the drug resistance promoting effect of miR-26b on PANC-1 .Conclusion miR-26b inhibits the expression of p53 by targeting 3′UTR of the p53 gene and enhances the drug resistance of pancreatic cancer cell line PANC-1 to gemcitabine .

Objective To study the expression level and function of micro RNA (microRNA)-218 in hepatocellular carcinoma (HCC) .Methods 46 cases of HCC surgery in the hepatobiliary surgery department of this hospital were selected and divided into the transfection group and nontransfection group .The expression ,proliferation and apoptosis of microRNA-218 and the expression level of B cell specific Maloney leukemia virus insertion site 1(Bmi-1) and cycling-dependented kinase 6(CDK6) in HepG2 cells were compared between the two groups .Results The expression level of microRNA-218 in HCC tissue was significantly lower than that in paracancerous tissues (P＜0 .05);the microRNA218 expression level was closely correlated with the clinicopathological characteristics such as tumor size and TNM stage(P＜0 .05);the HepG2 cell proliferation rates at 24 ,48 ,72 h after transfection in the transfection group were significantly lower than those in the nontransfection group(P＜0 .05);the HepG2 cell apoptosis rate in the transfection group was significantly higher than that in the nontransfection group(P＜0 .05);the Bim-1 and CDK6 expression levels after HepG2 cell transfection in the transfection group were significantly lower than those in nontransfection group(P＜0 .05) . Conclusion microRNA-218 can suppress the proliferation of HCC cells and promotes HCC cells apoptosis by down-regulating the Bim-1 and CDK6 expression level in potential targets .

Objective To explore the relationship between serum soluble human matrix lysine 2 (sST2) with chronic heart failure(CHF) and its clinical value for diagnosis and prognosis of CHF .Methods 60 cases of CHF and 60 cases of non-CHF were selected as the CHF group and control group respectively ,and the CHF group was divided into sST2 low level group and sST2 high level group according to the diagnostic threshold .The ELISA method was used to detect the serum sST 2 level of each group .The CHF group were followed up for 6 months .Then the influence of sST2 on CHF prognosis survival rate was observed .Results There was no statistical difference in age ,gender ,body mass index ,basic disease history ,basic medication situation and blood lipid indexes between the CHF group and control group(P＞0 .05);serum brain natriuretic peptide(BNP) level in the CHF group was obviously higher than that in the control group(P＜0 .01);serum sST2 levels in the CHF group and control group were (55 .08 ± 3 .98)ng/mL and (10 .46 ± 0 .72)ng/mL ,the difference was statistically significant (P＜0 .01) .Serum sST2 was positively correlated with BNP(r=0 .4606 ,P＜0 .01) ,moreover 95% CI was 0 .3066-0 .5911 .When the critical value was 0 .5303 ,the area under curve ,95% CI ,sensitivity ,specificity and likelihood ratio of sST 2 combined BNP detection were 0 .9362 ,0 .8853 -0 .9877 , 85 .00% (73 .43% -92 .90% ) ,98 .33% (91 .06% -99 .96% ) and 50 .00 respectively .The survival curve had statistical difference between the sST2 low level group and sST2 high level group(P=0 .0149) .Conclusion Serum sST2 can be used as a new biomarker for the diagnosis and prognosis of CHF ,and its combined with BNP may have better diagnostic value.

Objective To investigate the change of serum pigment epithelium derived factor (PEDF) ,β amyloid (Aβ) ,homocysteine (Hcy) and β endorphin (β-EP) levels in the patients with primary glaucoma and their clinical value .Methods 93 cases of primary glaucoma and 93 healthy people from July 2011 to June 2015 were selected as the primary glaucoma group and control group respectively .The ultrasound biomicroscopic examination results served as the golden standard .The changes of PEDF ,Aβ,Hcy and β-EP levels were observed in the two groups .Their sensitivity ,specificity and accuracy were compared .Results The PEDF level in the primary glaucoma group was significantly lower than that in the control group ,while serum Aβ,Hcy and β-EP levels were significantly higher than those in the control group ,the differences were statistically significant (P＜0 .05) .In the Richardson stage of primary glaucoma severity ,the higher the grade ,the higher the serum Aβ,Hcy and β-EP levels ,while the lower the PEDF level ,the differences were statistically significant (P＜0 .05) .The sensitivity ,specificity and accuracy of serum PEDF ,Aβ,Hcy and β-EP combined detection were 97 .26% ,90 .00% and 95 .70% respectively ,which were significantly higher than that of single index detection or other indexes combined detection(P＜0 .05) .Conclusion The levels of serum PEDF ,Aβ,Hcy and β-EP are higher in the patients with primary glaucoma and their combined detection has a great value for evaluating the severity of primary glaucoma .

Objective To research the clinical value of cerebrospinal fluid and peripheral blood tuberculosis infection T cell spot test(TSPOT .TB) in the diagnosis of tuberculous meningitis(TBM) .Methods 35 cases of TBM and 40 cases of non-TB intracranial infection in this hospital from September 2015 to September 2016 were selected as the observation group and control group respectively .T-SPOT .TB was adopted to detect mycobacterium tuberculosis effector T cells in cerebrospinal fluid and detect peripheral blood mononuclear cells .Results The T-SPOT .TB detection positive rates of cerebrospinal fluid and peripheral blood in the observation group were 97 .14% and 80 .00% respectively ,which were significantly higher than 2 .50% and 0 .00% in the control group ,the difference was statistically significant(P＜ 0 .05);the sensitivity and negative predictive value in cerebrospinal fluid T-SPOT .TB detection were 97 .14% and 97 .50% ,which were significantly higher than 80 .00% and 85 .11% in peripheral blood T-SPOT .TB detection ,the difference was statistically significant (P＜0 .05);the specificity and positive predictive value in the cerebrospinal fluid T-SPOT .TB detection was 97 .50% and 97 .14% respectively ,while which in peripheral blood T-SPOT .TB detection were 100 .00% and 100 .00% respectively ,but the difference between the two groups was not statistically significant ( P＞0 .05) .Conclusion The detection of cerebrospinal fluid and peripheral blood T-SPOT .TB detection can provide early diagnostic basis for TBM ,moreover cerebrospinal fluid T-SPOT .TB detection has higher sensitivity and possesses an important clinical application value .

Objective To investigate the clinical application value of peripheral blood lymphocyte subsets expression levels in autoimmune disease(AID) among the patients with antinuclear antibody(ANA) positive .Methods 200 patients with ANA positive and 196 patients with ANA negative were selected as the experimental group and control group respectively .The experimental group adopted indirect immunofluorescence assay (IFA) and immunoblotting assay(LIA) for detecting ANA ,moreover divided into the IFA group and LIA group according to the detection results .Meanwhile the flow cytometry was adopted to detect peripheral blood T lymphocytes ,helper T lymphocytes ,cytotoxic T lymphocyte ,NK lymphocytes and B lymphocytes absolute values of each group .The detection results were statistically analyzed .Results Helper T lymphocytes ,NK lymphocytes and B lymphocytes absolute values in the experimental group were significantly lower than those in the control group ,the differences were statistically significant(P＜0 .01);in the experimental group ,helper T lymphocytes and NK lymphocytes absolute values in the IIF group were significantly lower than those in the LIA group ,the differences were statistically significant (P＜0 .01) .Conclusion Peripheral blood lymphocytes subsets can serve as the important detection indicators during the diagnosis and treatment process of AID .

Objective To investigate the correlation of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphism with coronary atherosclerotic heart disease(CHD) risk factors in Yancheng area .Methods A total of 124 cases of CHD and 163 persons undergoing physical examination served as the CHD group and CON group .The enzyme method was adopted to detect TG and TC .The direction method was adopted to detect HDL and LDL .ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphisms were detected by adopting RFLP-PCR .Results The ratios of smoking history ,family history ,complicating hypertension and diabetes ,and the level of body mass index ,TC ,TG and LDLC in the CHD group were significantly higher than those in the control group ,the difference was statistically significant (P＜0 .05) .The various indicators had no statistically difference between male and female(P＞0 .05) .The frequency distribution and geographic distribution of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,Alu Ⅰ digestion polymorphisms had no difference between the two groups ,all conformed to Hardy-Weinberg genetic equilibrium and had the group representativeness .pp ,xx ,RR and AA genotypes in the CHD group were maximal ,while PP , XX ,rr and aa genotypes were minimal ;Pp ,xx ,RR and AA genotypes in the CON group were maximal ,while PP ,XX ,rr and aa genotypes were minimal .The distribution frequency of p and x genes in the CHD group was significantly higher than that in the control group ,the difference was statistically significant(P＜0 .05) .Conclusion The estrogen gene polymorphism might be a target spot for effectively treating CHD ,and p and x gene distribution frequency may be related with CHD risk factors .

Objective To explore the causes of Fra-1 high expression in breast cancer cells and molecular mechanism of Fra-1 high transcription activity .Methods Two human breast cancer cell lines MDA231 and MCF-7 and 1 human umbilical vein endothelial cell ECV304 were selected as the research objects .Real time fluorescent quantitative reverse transcription -polymerase chain re-action and Western blotting were used to detect the Fra-1 mRNA and protein expression ;the Fra-1 dual fluorescence reporter vector was constructed ,and the Fra-1 promotor transcription activity was detected ;the Fra-1 promotor short-cut reporter vector and mutation reporter vector at related loci were constructed ,and the transcription activity of short-cut reporter vector and mutation reporter vector was detected ;the binding situation of specific probe marked by biotin with activator protein-1(AP1) was observed by using the gel migration block test .Results The relative expression of mRNA and protein and promotor whole length reporter vector transcription activity of Fra-1 in human breast cancer cell lines MDA231 and MCF-7 cells were higher than that in human umbilical vein endothelial cell MCV304 ,the difference was statistically significant (P＜0 .05);in constructed series Fra-1 promotor shortcut reporter vectors ,only the activity of pGL3B-256 was obviously decreased ,the difference was statistically significant (P＜0 .05);the activity of AP1 mutation reporter vector pGL3B-M2 and SP1 and AP1 dual mutation reporter vector pGL3B-M3 was significantly lower than that of wild repoter vector pGL 3B-M1 ,the difference was statistically significant (P＜0 .05);the gel migration block test found that the binding of specific probe marked by biotin with AP1 had obvious blocking effect .Conclusion In in vitro cultured breast cancer cell lines MDA231 and MCF-7 ,the trans-acting factor AP1 up-regulates its expression by activating Fra-1 gene promoter .