Objective To investigate the diagnostic value of determination of the genotypes in codon 12 and 13 of K-ras oncogene in blood samples of patients with pancreatic carcinoma(PC).Methods Blood samples were obtained from 54 patients with pathologically confirmed PC,and 33 healthy controls.The DNAs were obtained in these samples.and then genotype of K-ras mutation was detected by using the PNA-clamping real-time quantitative PCR.Then the correlation between the K-ras genotypes of blood DNA and the clinical characteristics was analyzed.Results K-ras mutations were found in 74.1%(40/54)of patients with PC.There was no such mutation in control samples.The mutations of K-ras was associated with age,lymph node and vessel invasion.poorly differentiated tumor,CA19-9,while it was not associated with sex,tumor location,size of tumor,clinical staging and pathological type.Conclusions The one-step method was highly sensitive for detecting K-ras mutation in blood samples.Detection of circular blood cells harboring K-ras mutation suggested the tumor was highly invasive with poor prognosis.

The O152 antigens of Escherichia coli contains a Glc-β-1,3-GlcNAc linkage within the repeating unit. The wfgD gene in E. coli O152 O antigen gene cluster had been demonstrated utilizing NMR technique to encode a glucosyltransferase which is responsible for the synthesis of Glc-β-1,3-GlcNAc linkage. In this study, a synthetic substrate analog of the natural acceptor substrate undecaprenol-pyrophosphate-lipid [GlcNAc-α-PO_3-PO_3-(CH_2)_(11))-O-phenyl]) was used as an acceptor and UDP-Glc as a donor substrate, and electrospray ionization tandem mass spectrometry(ESI-MS-MS) was used for the detailed structural characte-)rization) of the enzyme product. A systematic study was conducted on product to allow rationalization of the fragmentation) processes. The major fragments observed in the ESI-MS-MS spectra result from cleavage of glycosidic) bond and diphosphate moiety. The fragment originating from the nonreducing end of the product yields information on sequence). Cross-ring cleavages, which are very informative of the linkages of the monosaccharide residues constituting) the product, and "internal" cleavage ions which are derived from elimination of substituents from around) the pyranose ring, were also observed. This extensive fragmentation was shown that the expected Glc-β-1,3-GlcNAc linkage in the product, confirming that wfgD is in the form of UDP-Glc: GlcNAc-pyrophosphate-lipid β-1,3-glucosyltransferase.)

Objective To explore the possible effect and mechanism of quercetin (QU) in inhibiting scar formation after the alkali burn of rat's cornea. Methods We established corneal alkali-burn model on right eyes of the SD rats. The rats were divided into five groups randomly. The control group received blank ophthalmic gel; the QU treatment groups received 2.5, 5, 10, or 20g/kg quercetin ophthalmic gel, respectively. The rats were checked by slit-lamp microscope every day for the degree of corneal opacity, then were killed on day 4, 7, 14, 21 and 28 after operation. The infiltration of the inflammatory cells was observed by histology, the arrangement and proliferation of the collagen fibers in the corneal stroma were observed by Masson trichrome staining. The expression of transforming growth factor-β_1 (TGF-β_1) and receptor of transforming growth factor-βⅠ(TGF-βRⅠ) in cornea was observed by immunohistochemical method. Results The corneal opacity was less severe in the QU treatment groups than in the control group (P<0.05). Corneal scar was inhibited better in 10g/kg QU and 20g/kg QU groups than in 2.5g/kg and 5g/kg QU groups, and 10g/kg and 20g/kg QU groups had no significant differences (P>0.05). HE and Masson staining showed that the density of corneal stroma collagen fibers in the QU treatment groups was lower than that in the control group. The expression of TGF-β_1 and TGF-βRⅠ reached the peak on day 7 after alkali burn, and then decreased slowly, which was close to the normal level. The expression of TGF-β_1 and TGF-βRⅠ was inhibited better in 10g/kg and 20g/kg QU groups than in 2.5g/kg and 5g/kg groups, and 10g/kg and 20g/kg QU groups had no significant differences (P>0.05). Conclusion QU ophthalmic gel can reduce formation of corneal scar to a certain extent, 10g/kg is the optimal concentration. QU may play its role by inhibiting the expression of TGF-β1 and TGF-βRⅠ.

Objective To investigate the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling pathway in the endometrium of women with polycystic ovary syndrome (PCOS) and its effect and significance in the cause of hyperplasia and carcinoma;and investigate the factors which affect the activation of the MAPK/ERK signaling pathway. Methods Collected 52 patients diagnosed as PCOS who were taken dilation and curettage of uterus as study, while 32 non-PCOS patients matched as control group. Serum hormonal parameters, fasting blood glucose and insulin were measured in all patients. The PCOS patients were sub-group as insulin resistance group and non-insulin resistance group; all the patients were carried out pathology inspection of endometria, and the PCOS patients were sub-group as endometrial hyperplasia and carcinoma group and normal endometrium group based on the outcome of pathology inspection. Western blot were performed to detect the expressions of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2), the activation of ERK1/2. Results (1)The expression of pERK1/2 [(61 ±13)%] in the endometrium in PCOS group was higher than that in the control [(44 ±10)%, P ＜0.01]. (2)The expression of p-ERK1/2 was significantly increased in group of endometrial hyperplasia and carcinoma [ ( 70 ± 11 )% ] compared to that in group of normal endometrium [ (55 ± 10)% ,P ＜ 0.01 ], while there were significant difference between group of insulin resistance [ (63 ± 13 )% ] and group of non-insulin resistance [ (55 ±7)%, P ＜0.01 ]. (3) Fasting insulin level, insulin area under the curve and body mass index were related to the expression of p-ERK1/2 in endometrium with PCOS, the correlation coefficient were 0.447, 0.456 and 0.381, respectively ( all P ＜ 0.01 ). Conclusions The MAPK/ERK signaling pathway in endometrium with PCOS was overactivation, which was related to the endometrial hyperplasia and carcinoma; while the activation of MAPK/ERK signaling pathway were effected by insulin resistance and hyperinsulinemia.

Objective To investigate the effect of recombinant human growth hormone (rhGH) action on pregnancy rate of polycystic ovarian syndrome(PCOS).Methods The PCOS who were client in our hospital in February-August,2012 were investigated.The rhGH group (41 cases) were that all patients took rhGH at the same time ovulation.The control group (41 cases) were no medication group.The two groups had sex action after ovulation.Ovulation rate,pregnancy rate and endometrial score of the two groups were compared.Results Ovulation rate of rhGH group was 90.2％ (37/41),the control group was 68.2％ (28/41),the difference was statistically significant (x2 =3.459,P =0.036).Clinical pregnancy rate of RhGH group was 32.7％ (12/37),the control group was 25.0％ (7/28).The difference was statistics significance (x2 =4.319,P =0.032).Endometrial score of rhGH group was (14.96 ± 2.18),the control group was (12.77 ± 2.60) points,the difference between groups was statistically significant (t =-3.356,P =0.032).Conclusion Giving rhGH ovulation cycle specification can promote ovulation PCOS patients and improve the adverse factors affecting pregnancy,improving the pregnancy rate.

Objective To investigate the changes of zinc in maternal plasma and MMP-9,collagen Ⅳ levels in serum and matrix metalloproteinases-9 (MMP-9) level in amniotic fluid in patients with premature rupture of membranes at term(TPROM).Methods Thirty cases who were diagnosed as PROM at term during Nov.2012 to Mar.2013 were enrolled as case group,and 30 cases delivered during the same time without PROM were enrolled as control group.Maternal blood and ammiotic fluid were collected from all the cases.The level of zinc in maternal plasma was measured by atomic absorption method and the levels of MMP-9 in serum and amniotic fluid were detected by enzyme linked immunosorbent assay(ELISA),while the level of collagen Ⅳ in serum was measured by up-conversion luminescence method.The relationship among them was analyzed.Results Compared to control group,there were statistically significant difference between TPROM and control groups in terms of the level of zinc,collagen Ⅳ,MMP-9 in serum and MMP-9 in amniotic fluid (zinc:(109.10 ± 16.07) μmol/L vs.(90.54 ± 10.99) μmol/L; t =-5.22,P ＜ 0.001 ; collagen Ⅳ:(56.86 ±41.26) μg/L vs.(88.61 ±44.87) μg/L;t =2.852,P =0.006;MMP-9 in serum:(1 463.25 ±483.6) μg/L vs.(1 196.9 ± 357.43) μg/L,t =-2.426,P =0.018 ; MMP-9 in amniotic fluid:(125.48 ± 67.18) μg/L vs.(72.64 ± 60.74) μg/L,t =-2.873,P =0.006).Zinc level in maternal plasma and collagen Ⅳ in serum had a negative relationship in TPROM (r =-0.261,P =0.044).Zinc level in maternal plasma and MMP-9 level in serum had a positive relationship in TPROM (r =0.274,P =0.034).MMP-9 levels in serum and amniotic fluid had a positive relationship in TPROM (r =0.264,P =0.047).There were no significant relationship between zinc level in maternal plasma,MMP-9 level in amniotic fluid,collagen Ⅳ and MMP-9 levels in serum,collagen Ⅳ in serum and MMP-9 in amniotic fluid (r =0.215,-0.172,-0.172 ; P ＞ 0.05).Conclusion The level of zinc in maternal plasma and increase of MMP-9 in serum and amniotic fluid of women and decrease of the level of collagen Ⅳ in serum are related to the occurrence of TPROM.

Objective To investigate the relationship between the onset of lower extremity venous thrombotic disease and seasons.Methods A retrospective study was conducted, 300 patients with lower extremity venous thrombotic disease admitted to HandanCity Hospital of Traditional Chinese Medicine (TCM) from August 2012 to February 2014 were enrolled, the incidences and TCM syndromes of patients with lower extremity venous thrombotic disease in different seasons were observed, and the pathogenesis and relationships between the types of TCM syndrome and seasons were analyzed.Results There were 142 patients with lower extremity superficial thrombophlebitis, and 158 cases with lower extremity deep venous thrombosis, the incidence of lower extremity venous thrombotic disease in spring was significantly higher than that in summer and autumn [32.8% (86/262) vs. 21.3% (54/254), 18.4% (50/272), bothP < 0.01], but lower than that in winter [32.8% (86/262) vs. 37.2% (110/296)], the difference was not statistically significant (P > 0.01); while the incidence of lower extremity venous thrombotic disease in winter was significant higher than those in summer and autumn (allP < 0.01). The incidences of damp and heat downward flow type in autumn and summer were increased compared with those in winter and spring [55.6% (30/54), 60.0% (30/50) vs. 20.0% (22/110), 23.3% (20/86), allP < 0.01], while the incidences of lower extremity venous thrombotic disease with damp heat and stasis syndrome in winter and spring were increased compared with those in summer and autumn seasons [80.0% (88/110), 76.7% (66/86) vs. 44.4% (24/54), 40.0% (20/50)].Conclusions The incidence of lower extremity venous thrombotic disease is related to seasons, and the onset is high in winter and spring, damp heat and stasis syndrome being the main type; according to different seasons, clinical treatment can direct to different pathogenic factors to adopt different preventive measures interfering with the patient's constitution in order to eliminate or reduce the risk factors, achieving the effect of the disease prevention.

N)compared with B/HongKong/330/2001.Conclusions H1,H3 and B virus were circulated in Hebei from 2005 to 2006.Recent viruses were changing in genetic characteristics,while influenza B viruses varied more obviously.

Objective To understand the genetic characterization of HA1 gene of influenza A H3N2 viruses circulated in recent years in Hebei.Methods Viral RNAs of 25 H3N2 strains were extracted and amplified by Reverse Transcription Polymerase Chain Reaction (RT-PCR).The products of PCR were purified and sequenced,and the sequences were analyzed though biometic software.Results Several amino acid substitutions located in antigenic sites or receptor binding sites were found more in the isolates than the current vaccine virus or the isolates from previous year.Amino acid substitution was found in 3,140,142,144,145,158,159,189,192,193,198,204,225,226 and 227 positions in the isolates during 2003-2008,more amino acid substitutions took place in antigenic determinant A,B and receptor binding site (RBS).New phylogenetic branches appeared continuously during 2003-2008.The H3N2 strains of the same year almost clustered in the same group on the phylogenetic tree.Conclusions Amino acid substitutions continuously occurred in the HA1 genes in influenza A H3N2 viruses isolated in Hebei from 2003 to 2008,it is meaningful to pay close attention to the HA1 variation in order to prevent and control influenza.

Objective To explore the role and their relationship of Ki-67, VEGF and p27 expressed in adult patients with acute leukemia. Methods The expression of Ki-67, VEGF and p27 in bone marrow mononuclear cells (BMMNC) were analyzed by immunocytochemical staining, and their correlations of Ki-67, VEGF and p27 were statistically analyzed. Results The expression of Ki-67(42.48±25.78)% or VEGF (44.89±24.01)% on BMMNC from acute leukemia cells was significantly higher than that in the control (11.40±9.94)% or (16.90±12.54)% (P<0.01). But the expression of p27 (23.65±13.30)% was significantly lower than that in the control (50.23±22.68)% (P<0.01). The expressions of Ki-67 were positively correlated with and VEGF in patients with acute leukemia were positively correlated(r=-0.666, P<0.01), and the expressions of Ki-67 and p27 were negatively correlated with p27 in patients with acute leukemia (r=-0.316, P<0.05).Conclusion The evaluation of expression of Ki-67, VEGF and p27 on acute leukemic cells provides new insights to the pathogenesis is helpful in mechanism and is helpful in the diagnosis of acute leukemia.