Objective To study the early diagnosis of heamolytic disease of newborn(HDN) by umbilical blood immunoserological test. Methods According to the laboratory rules, immunoserological tests were practised with neonatal umbilical blood of 916 A or B blood-type neonates delivered by O-type blood mothers. The morbidity and the pathogenic time of hyperbilirubinemia were studied in 7 days after birth to the neonates whose erythrocyte was sensitized by the anti-A(B) IgG. Results (1) There was significant difference in the morbidity of hyperbilirubinemia of the neonates whose umbilical blood antibody-released test results were postive compared with control group (P<0.01), There had no significant difference in the morbidity of hyperbilirubinemia of the neonates compared with puerperal times. (2) There had significant difference in the morbidity of hyperbilirubinemia between the neonates having free antibody and those without free antibody (P < 0.01). (3) The peek time of hyperbilirubinemia was in 1-3 days after birth in patient-infants whose antibodyreleased test result was positive. Conclusion The umbilical blood immunoserological test is an exact method of the early diagnosis to HDN. The free antibody helps to estimate the developing trend of HDN.

Objective To discuss the determination of estriol and placenta prolactin levels for fetal grow th and development and provide theoretical basis for doctors to improve the rate of eugenic and superior nurture .Methods The pregnant women treated in the hospital from August 2014 and February 2015 were divided into two groups :pregnant women with fetal growth restriction were as the observation group(n=59) ,who with normal fetal development were as control group(n=256) .Estriol and placenta prolactin levels of two groups tested ,and the data obtained were statistically analyzed .Results Estriol level of the observation group was (13 .36 ± 2 .57)ng/mL ,which of the control group was (21 .34 ± 3 .48)ng/mL ,and the level of the observation group was lower than the control group significantly(P<0 .05) .Placenta prolactin concentration of the observation group was (3 .28 ± 0 .84)ng/mL , taht of the control group was (5 .27 ± 1 .24) ng/mL ,and the level of the observation group was lower than the control group signifi‐cantly(P<0 .05) .Estriol positive rate of observation group was 40 .68% (24/59) ,placenta prolactin positive rate was 37 .29%(22/59) ,and the positive rate of combined detection was 71 .19% (42/59) .Conclusion Examination the estriol and placenta prolac‐tin levels of pregnant female can evaluate fetal placental function .

Objective To investigate the serum sex hormone changes and fertility condition in childbearing age women patients with hyperthyroidism treated by iodine-131 .Methods 50 cases of healthy volunteers were selected .248 childbearing age women pa-tients with hyperthyroidism were divided into the iodine-131 group and the anti-thyroid medication group according to the treatment method .The thyroid function and gonad hormone before treatment and at 6 months after treatment were detected in 3 groups ,and the pregnancy and fetal condition were tracked .Results The change of thyroid stimulation hormone(TSH) level was more sensitive in the iodine-131 treatment group than the anti-thyroid medication group ;luteinizing hormone(LH) ,follicle stimulation hormone (FSH) ,prolactin(PRL) ,testosterone(T) and progesterone(P) before treatment in the two groups were significantly increased com-pared with the control group ,but E2 was decreased ;the changes of sex hormones in the iodine-131 treatment group had differences between before and after treatment ,the changes of T and P in the anti-thyroid medication group were unobvious before and after treatment ;the normal delivery rate in the control group and the iodine-131 therapy group had no statistical difference (χ2 =0 .148 , P=0 .7>0 .05) ,the normal delivery rate in the control group and the anti-thyroid medication group had statistically significant difference(χ2 = 5 .739 ,P=0 .017<0 .05) ,the normal delivery rate in the iodine-131 therapy group and the anti-thyroid medication group had statistically significant difference(χ2 =4 .26 ,P=0 .039<0 .05) .Conclusion The iodine-131 therapy has no influence on the sex hormones and the fertility ability in childbearing age women patients with hyperthyroidism ,and also does not increase the incidence rate of genetic damage .

Objective:To determine the expression of BMP4 in hepatocellular carcinoma (HCC) and to study the role of BMP4 in inducing epithelial-mesenchymal transition (EMT) to analyze the effect of BMP4 on the migration and invasion of HCC cells. Methods: The expression of BMP4 in HCC specimens was examined by immunohistochemistry staining, and the correlations were analyzed between the expression of BMP4 and clinicopathological data. The BMP4 expression plasmid was transfected into HepG2 cells to induce exogenous overexpression of BMP4 protein. The changes of HepG2 cell morphology were detected after BMP4 transfection by using a microscope; the changes of the expression of BMP4, EMT-related protein (E-cadherin, Vimentin) in HepG2 cells were detected by Western blot after transfection of BMP4;the wound healing assay in vitro was used to detect the effects of BMP4 gene transfection on the ability of migration of HepG2 cells;the invasion assay was used to determine the role of transfection of BMP4 on the invasive potential of HepG2 cells. Results: Immunohistochemistry staining method displayed that BMP4 expression was positively associated with age, histological differentiation, stage, and poor prognosis. After BMP4 overexpression, the morphology of HepG2 cells showed significant changes from a paving stone structure with cell-cell adhesion to a fibroblastic shape, which showed typical EMT change; Western blot exhibited that the expression of E-cadherin was downregulated and the Vimentin expression was upregulated in HepG2 cells;the wound healing and invasion assay showed that the migration and invasion potentials of HepG2 cells were significantly enhanced. Conclusion: BMP4, which displayed a high expression in HCC specimens, was closely associated with clinicopathologic data, and BMP4 may promote migration and invasion of HCC cells by inducing epithelial-mesenchymal transition.

Objective: To investigate differentially expressed protein profiles in B16-F10 grafted melanoma and its metastasis in the lung in order to identify molecular markers of melanoma metastasis. Methods: Differentially expressed proteins in B16-F10 grafted melanoma and its metastatic lesion in the lung were isolated and identified by fluorescence two-dimensional differential gel electrophoresis(2D-DIGE)coupled with matrix assisted laser desorption ionisation time-of-flight mass spectrometry(MALDI-TOF-MS).Some of identified proteins were further confirmed by Real-time PCR analysis. Results: High resolutional images of differential gel electrophoresis were obtained and 9 of 30 differentially expressed proteins (IRatiol≥2,P<0.01)were identified by MALDI-TOF-MS.The expression of Myoglobin(MB),vimentin(VIM),phosphoglycerate kinase 1(PGK1),Triosephosphate isomerase(TPI or TIM),heavy-chain binding protein(BiP),α-enolase,β-actin,γ-actin,and laminin-binding protein were up-regulated in the experimental group compared with the control group.These proteins were involved in the cytoskeletal formation,glycolysis and so on.Real-time PCR analysis showed up-regulation of mRNA expression of PGK1 and TPI in the experimental group(P=0.001 and 0.003),which was in consistent with the resuits of proteomic analysis. Conclusion: A variety of abnormally expressed proteins contribute to the metastasis of mice melanoma.Glycolytic enzymes PGK1 and TPI may be involved in this process.

Objective: To investigate the influence of TGF-β and IFN-γ on the proliferation, migration and invasion of melanoma cells. Methods: Melanoma cells were cultured in vitro. When tumor cells were confluent about 80% degree, cytokines were added into cell culture media. The concentration of TGF-β and IFN-β was 5ng/mL and 10ng/mL, respectively. Melanoma cells were divided into free-cytokine group, TGF-β group,IFN-γ group, TGF-β and IFN-γ group. Tumor cells in each group were then incubated for 8h, 16h, 24h, 32h,40h and 48h, respectively. After incubation, fixing and staining with SRB, the optical densities and percentage viability were then determined by absorption at 540 nm (A 540). The scarification of tumor cells in each group on the surface was created by a 2001μL pipette tube. The motility of tumor cells in each group was assessed by measuring the distance between scarifications. The speed of the scuffing closure was monitored after 12h.The invasive ability of melanoma cells was observed by transwell cultivation. The tumor cells that invaded through the Matrigel and adhered to the bottom of the outside membrane were determined by absorption at 595 nm (A595). Gelatin zymography assay was used to examine the levels of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) activity when the tumor cells were treated with cytokines after 24h. MMP-2 and MMP-9 activity was demonstrated by gradation in the sodium dodecyl sulfate polyacryl-amide gel electrophoresis (SDS-PAGE) gelatin. MMP-2 and MMP-9 activity was determined by Image analy-sis Software. Results: TGF-β promoted the proliferation, migration and invasion of melanoma cells (P<0.05).However, IFN-γ inhibited the proliferation, migration and invasion of melanoma cells (P<0.05). The effect weakened or disappeared when both of them were used (P>0.05). Conclusion: In vitro, TGF-β may affect the inhibitory effect of IFN-γ on the proliferation, migration and invasion of melanoma cells. This study provided a better understanding of the relationship between tumor and inflammatory factors and established a good ba-sis for future research.

Objective:To study the clinicopathologic characteristics and the prognostic factors of triple negative breast cancer,and to study the correlation between clinicopathologic features and EGFR expression.Methods:Clinical characteristics and prognosis of 200 breast cancer patients diagnosed between 1997 and 2004 were reviewed.The paraffin embedded tissues were selected and tissue microarray blocks were made.Immunohistochemical staining was used to evaluate the expression of ER,PR and HER2 for breast cancer molecular type.EGFR expression was evaluated to analyze its correlation with the clinical characteristics and prognosis.Results:Fourty-two cases(21%)of 200 were triple negative breast cancer(TNBC).There was no significant difference between TNBC and non-TNBC in clinicopathologic parameters such as age,tumor size,clinical stage,and lymph node status.However,there was significant difference in histological category and menostasis status between the two groups(P<0.05).Among TNBC,17 0f 42 died during the follow-up(the overall suivival rate was 59.52%),while 26 0f 158 non-TNBC died(the overall survival rate was 83.54%).EGFR was significantly overexpressed in TNBC(69.05%)and it was associated with lymph node status and histological category.Conclusion:Although the incidence of TNBC is low,it has a poorer prognosis than other subtypes.EGFR is highly expressed in TNBC and may serve as a prognostic index and an important therapatic target for breast cancer.

Objective To evaluate the diagnostic value of sympathetic skin response(SSR)in patients with vestibular vertigo.Methods SSR tests were performed on 1 20 patients with acute vestibular system vertigo,including 70 cases of central vertigo and 50 eases of peripherM vertigo.60 healthy subjects were also examined to serve as controls.Results In those with central vertigo,the abnormality rate in the SSR results was 87.1%(61/70).SSR latency was longer and its amplitude wag lower than in those with peripheral vertigo and in the heMthy controls.In those with peripheral vertigo the abnormality rate was 18.0%(9/50),but the average latency and amplitude were not significantly different from those of the healthy controls.Conclusion Persons with acute central vestibular vertigo may have sympathetic nerve dysfunction.SSR test results can be used as an electrophysiological index to distinguish central from peripheral vestibular vertigo.

Objective:To investigate the correlations of Lauren classification and world health organization (WHO) classification of gastric cancer (GC) with microvascular density (MVD), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2), and p53. Methods:The clinical data of 89 patients with GC were collected. The collected specimens were categorized on the basis of Lauren classification and WHO classification. CD34/periodic acid-Schiff (PAS) double staining was performed to validate MVD. Immunohistochemistry was conducted to investigate the expression levels of MMP-2, MMP-9, VEGF, VEGFR1, VEGFR2, and p53. Results:MVD was not correlated with Lauren classification or WHO classification (P>0.05). Lauren typing was associated with the expression levels of MMP-9, VEGFR1, and p53 (P<0.05). WHO classification was not related to any of the factors (P>0.05). Cox proportional hazards model revealed that Lauren classification and WHO classification were the prognostic factors of overall survival (P<0.05). Conclusion:This research on tumor related factors, angiogenesis, and different classifications of GC may provide new methods to treat this disease.

Objective:To investigate the clinical significance of epithelial-to-mesenchymal (EMT) in lung squamous cell carcino-ma (LSCC) and to examine the effect of EMT on the invasive and migration abilities of LSCC. Methods:Immunohistochemical stain-ing was performed to determine the expression of E-cadherin, Vimentin, and TGF-β1 in 79 LSCC patients, and the clinical significance was explored. SK-MES-1 lung squamous carcinoma cells were cultured in conditioned medium containing various concentrations of transforming growth factor-β1 (TGF-β1) for 5 and 10 days. The expression levels of E-cadherin and Vimentin were detected via West-ern blot and reverse transcription-polymerase chain reaction (RT-PCR). With different concentrations and induction times, invasion and wound healing assays were performed to evaluate the invasion and migration abilities. Results:E-cadherin expression was significantly lower, whereas Vimentin expression was significantly higher in LSCC with lymph node metastasis than in that without noda metastasis (P<0.05). In the tissues of 79 LSCC patients, TGF-β1 expression was significantly related to lymph node metastasis (P<0.05). Western blot showed that Vimentin expression was higher, whereas E-cadherin expression was lower in TGF-β1 inducing medium with 10 ng/mL SK-MES-1 cells than in the other media. RT-PCR showed similar results. Scratch test and invasion assay both showed that treat-ment of cells with cytokines markedly enhanced the migration and invasion of the cells. Conclusion:Lymph node metastasis of LSCC correlates with EMT. SK-MES-1 cells undergo EMT via TGF-β1 induction, which enhances invasion and migration.