Objective To establish blood screening and genetic detection of thalassemia trait in pregnant couples in Chang‐shou area so as to provide guidance for aristogenesis and prenatal diagnosis .Methods A total of 1 760 pregnancy in maternal and child health hospital treated from January 2013 to October 2014 were selected for study .The component of hemoglobin was ana‐lyzed as primary screening and genotype of pregnant couples were ensured in which primary screening result is positive .Results There were 27 cases suspected as α‐thalassemia (positive rate was 1 .53% ) and 25 cases suspected as β‐thalassemia(positive rate was 1 .42% ) in the primary screening(n=1 760) .The positive rate of gene carrier were 31 .51% (n=438) in women and 33 .33%(n=27) in men .Conclusion The routine screening of thalassemia could guide aristogenesis in high incidence area and provide terms of prenatal diagnosis and genetic counseling .

Objective To investigate the clinical significa nc e and the relationship between the serum squamous cell carcinoma antigen (SCC-A g) levels and the biological characteristics in patients with cervical carcinoma . Methods The pre-post-treatment sera from 500 patients w ith cervical carcinoma from 1998 to 2002 were analyzed for the SCC-Ag levels by IMX; and the correlation between the SCC-Ag level and the clinicopathologic ch aracteristics were also detected. Results Significant corre lation was found between the pre-treatment SCC-Ag level and pathologic classif ication, and clinical stage (P0.05); The pre-treatm ent SCC-Ag level is significantly higher than that of post-treatment (P

Objective To explore the value of the apparent diffusion coefficient (ADC)of MRI diffusion-weighted (DW)in diag-nosing pathologic stage and histologic grade of bladder cancer.Methods 42 patients with confirmed bladder cancer underwent pelvic MRI examination including T2 WI and diffusion-weighted imaging (b values of 0 and 1 000 s/mm2 )Based on pathologic results,42 cases of bladder cancer were divided into three groups of Ta-T1 stage (non-muscle invasive),T2 stage (muscle invasive)and T3-T4 stage (around the bladder and other organizations invasive).Of 42 bladder cancers,36 urothelial carcinomas were divided into papillary urothelial neoplasm with low malignant potential (PUNLMP),low grade urothelial carcinoma and high grade urothelial carcinoma group.Tumor diameter and ADC were measured on DW imaging,and compared among different groups.Results The mean size of high grade tumors (4.52 ± 1.61)cm was significantly larger than that of PUNLMP (2.28 ± 0.51)cm and low grade tumors (1.69 ± 0.53)cm.The mean ADC of high grade tumors (0.83 ± 0.27)×10-3 mm2/s was significantly lower than that of PUNLMP (1.46 ± 0.30)×10-3 mm2/s and low grade tumors (1.17±0.11)×10-3 mm2/s (P<0.01).T3-T4 stage tumors showed significantly lower ADC (0.82±0.21)×10-3 mm2/s than T2 (1.01 ± 0.09)×10 -3 mm2/s and Ta-T1 stage tumors (1.24 ± 0.13)×10 -3 mm2/s (P <0.01).Conclusion DW ima-ging combined with conventional MR sequences is useful for displaying morphology of bladder cancer.DW imaging and ADC are use-ful in evaluation of pathologic stage and histologic grade of bladder cancer.

[Abstract] Objective To explore the relationship between Kruppel-like factor 4 (KLF-4) and E-cadherin in human peritoneal mesothelial cells (HPMCs), and the expression and function of KLF-4 in the animal model of peritoneal fibrosis induced by high glucose peritoneal dialysate. Methods Co-transfection in HPMCs with the plasmid of KLF-4 and the bind site or mutant in the promoter region of E-cadherin, and then the luciferase activity was measured of the each bind site and its matched mutants to estimate whether KLF-4 can combine with the bind site in the promoter region of E-cadherin; Chromatin immunocoprecipitation (CHIP) was exploited to verify if KLF-4 can combine with the bind site in the promoter region of E-cadherin; Real-time PCR and Western blotting were performed to detect the expression of E-cadherin at the bind site and matched mutants of b, d, f and g. Thirty SD rats were randomly divided into saline group, peritoneal dialysate group and experimental group (10 each). Rats in saline group were given intraperitoneal injection with 0.9% NaCl, in peritoneal dialysate group were given with 4.25% high glucose peritoneal dialysate, and in experimental group were given via tail vein with 4.25% high glucose peritoneal dialysate and the mixture of KLF-4 plasmid suspension containing ultrasound microbubble. To observe the peritoneal tissue thickness of the 3 groups of rats by Hematoxylin and Eosin staining. Masson trichrome staining was performed to detect the deposition of collagen fibers in peritoneal tissue of the 3 groups of rats. Immunohistochemistry was used to detect the expression level of KLF-4, E-cadherin, α-SMA and fibronectin (FN) in peritoneal tissue of the 3 groups of rats. Results Promoter luciferase reporter gene and CHIP results showed that KLF-4 can combine with the bind site in the promoter region of E-cadherin in HPMCs. Real-time PCR and Western blotting showed that KLF-4 can positively regulate the expression of E-cadherin. HE staining showed that the peritoneal tissue was obviously thickened in rats of peritoneal dialysate group [(105.91±12.0) μm] than in rats of saline group [(20.89±5.39) μm] and of experimental group [(23.05±6.07) μm] with statistical significance (P<0.05), but the difference between saline group and experimental group was not statistically significant (P>0.05). Masson staining showed that the deposition of collagen fiber significantly increased in peritoneal dialysate group (0.89±0.09) than in saline group (0.19±0.03) and experimental group (0.15±0.06) with statistical significance (P<0.05), but the difference between saline group and experimental group was not statistically significant (P>0.05). Immunohistochemistry results showed that the expressions of KLF-4 and E-cadherin were obviously lower in peritoneal dialysate group (0.27±0.09, 0.31±0.03) than in saline group (0.79±0.19, 0.83±0.13) and experimental group (0.85±0.11, 0.76±0.11) with statistically significant difference (P<0.05), but no significant difference existed between saline group and experimental group (P>0.05). In contrast, the expressions of α-SMA and FN were evidently higher in peritoneal dialysate group (0.83±0.09, 0.63±0.09) than in saline group (0.22±0.08, 0.30±0.07) and experimental group (0.19±0.05, 0.11±0.03) with statistically significant difference (P<0.05), but no evident difference existed between saline group and experimental group (P>0.05). Conclusion KLF-4 may positive regulate the expression of E-cadherin by combining with the bind site in the promoter region of E-cadherin, and inhibit the peritoneal fibrosis induced via high glucose peritoneal dialysate.

Objective To explore the role of Kruppel-like factor 4 (KLF-4) in phenotypic transition of human peritoneal mesothelial cells (HPMCs) induced via high glucose. Methods HPMCs were induced by 50mmol/L glucose for 72 hours, the expressions of epithelium-cadherin (E-cadherin), KLF-4, α-smooth muscle actin (-SMA), connective tissue growth factor (CTGF) and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. The treated cells were transfected with LVKLF-4 and inhibitor, the untreated cells were transfected with shRNA-KLF-4 and mimic. The mRNA and protein expressions of KLF-4, E-cadherin, α-SMA, CTGF and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. Results Real-time PCR showed that the expression of E-cadherin decreased and of α-SMA, CTGF and miRNA-143 increased, but of KLF-4 not changed in high glucose treated cells. Western blotting showed that the expression of KLF-4 and E-cadherin decreased. Upregulating KLF-4 increased the expression of E-cadherin, but decreased the expression of α-SMA and CTGF. Down-regulating KLF-4 decreased the expression of E-cadherin, but augment the expression of α-SMA and CTGF. Conclusion High glucose may induce the down-regulation of KLF-4 protein, and SRF- miRNA-143-KLF-4 signal pathway axis may be involved in the process of HPMC phenotypic transition.

Adult ; Female ; Hemangioma ; pathology ; Humans ; Mesoderm ; pathology ; Placenta ; pathology ; Placenta Diseases ; pathology ; Pregnancy ; Young Adult

Acquired Immunodeficiency Syndrome ; China ; Cross-Sectional Studies ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male

Objective To construct a recombinant expression vector for expression of the function-al domains of dengue virus serotype 1 ( DENV1 ) envelope ( E ) protein in native soluble form. Methods The genes encoding the functional domains of DENV1-E protein (1-394 aa) were amplified with PCR and then cloned into the Psectag2B-Fc eukaryotic expression vector.The 293T cells were transfected with the recombinant vector by cationic lipid-based delivery.The cell clones expressing the fusion DENV1-E-Fc protein were screened out with 2 mg/ml of Zeocin.Immunofluorescence assay ( IFA) was performed to analyze the antigenicity and integrity of the fusion protein.The fusion proteins were purified from cell lysate with Protein-G and further identified by Western blot assay.Results The soluble form of fusion protein with a molecular weight of about 90×103 was obtained at a yield of about 25 μg per 1×107 cells.The results of IFA indicated that the fusion protein kept its integrity with right conformational epitopes.The fusion protein was successfully expressed with the advantage of good specificity as indicated by IFA and Western blot assay. Conclusion The recombinant fusion protein in soluble form was successfully expressed in eukaryotic ex-pression system, which paved the way for further investigation on the function of DENV1 E protein and its protective epitopes.

Abstract?AIM: To investigate the effect of epigallocatechin-3-gallate ( EGCG ) against oxidative stress induced by high glucose in human lens epithelium ( HLE) cells.? METHODS: The HLE cell oxidative damage model induced by high concentration glucose was established, and was intervented with different concentrations of EGCG. Cell viability was determined by MTT assay, cell morphology was investigated by convert microscope, cells apoptosis was assayed by flow cytometry with Hoechst-PI staining. Moreover, the levels of super oxide dismutase ( SOD) , glutathione peroxidase ( GSH-Px) and malondialdehyde ( MDA) in supernatant were also tested after different treatment either with high concentration glucose or with different concentrations of EGCG.?RESULTS: MTT results showed that HLE cells activity increased to 50. 33%± 3. 52% and 63. 33%± 4. 63% after treated with 10 μmol/L and 100 μmol/L EGCG respectively, the difference was statistically significant compared with oxidative injury group(32. 67%±3. 10%)(P<0. 05 ); HLE cells maintained better morphology intervented with EGCG under high glucose conditions, the number of apoptotic cells reduced, SOD and GSH-Px level within HLE cells increased and MDA levels decreased.?CONCLUSION:EGCG plays its strong antioxidant effect by increasing SOD, GSH-Px content and decreasing MDA content in cells, therefore provides a reliable experimental basis for the search for effective prevention and treatment of cataract drug.

Objective To evaluate cytotoxic effects of cytokine-induced killer cells (CIK cells) transfected with the interleukin-2 (IL-2) gene on malignant melanoma cells.Methods Mouse spleen cells were extracted,lymphocyte cells were separated,and CIK cells were prepared from these lymphocyte cells.PEGF-N1 plasmids containing IL-2 gene (PEGF-NI-IL-2) were transfected into CIK cells.Fluorescence microscopy was used to observe transfection products,and reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to determine the IL-2 mRNA expression.Then,effector cells such as CIK cells and IL-2-transfected CIK cells were separately co-cultured with target cells (B16 melanoma cells) at effector-target ratios of 10∶ 1,20∶1 and 40∶1,then 4-hour lactate dehydrogenase release assay was performed to evaluate cytotoxic effects of the two kinds of CIK cells on B 16 cells.After effector cells were cocultured with target cells at the effector-target ratio of 40∶1 for 48 hours,enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-2,interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant of the two kinds of CIK cells.Finally,mouse models of melanoma were established,and a total of 28 melanoma-bearing mice were divided into 4 groups to be peritumorally injected with 0.2 ml sodium chloride physiological solution (control group),100 IU IL-2 solution (IL-2 group),CIK cell suspension at a cell density of 1 × 106 cells per milliliter (CIK group) and IL-2-transfected CIK cell suspension at a cell density of 1 × 106 cells per milliliter (IL-2-transfected CIK group) respectively.Tumor morphology,tumor inhibition rate and cell apoptosis rate were used to evaluate tumor growth in the above groups.If data were normally distributed,t-test was used for comparing means between two groups,and analysis of variance and least significant difference (LSD)-t test were used for comparing means among multiple groups.Results Fluorescence microscopy and RT-PCR both showed that IL-2 was successfully transfected into CIK cells.The cytotoxic effect of IL-2-transfected CIK cells on B16 cells was strongest at the effector-target ratio of 40:1.Levels of IL-2,IFN-γ and TNF-α were also significantly higher in the supernatant of IL-2-transfected CIK cells [(1107.26 ± 6.49) pg/ml,(50.01 ± 3.35) pg/ml,(39.86 ± 3.25) pg/ml] than those in that of CIK cells [(51.09 ± 3.85) pg/ml,(32.71 ± 2.43) pg/ml,(30.11 ± 3.08) pg/ml,t =442.60,14.93,6.89,all P ＜ 0.01].Animal experiments showed that the tumor volume obviously increased in the control group (P ＜ 0.05),but significantly decreased in the IL-2 group,CIK group and IL-2-transfected CIK group (all P ＜ 0.001) after intervention compared with those before intervention.Furthermore,the tumor volume in the IL-2-transfected CIK group was significantly less than that in the other three groups (all P ＜ 0.01),but no significant difference was observed between the IL-2 group and CIK group (P ＞ 0.05).In addition,the apoptosis rate was significantly higher in the IL-2 group,CIK group,and IL-2-transfeeted CIK group than that in the control group (all P ＜ 0.01).The apoptosis rate and tumor inhibition rate were significantly higher in the IL-2-transfected CIK group than those in the IL-2 group and CIK group (all P ＜ 0.01),but insignificantly different between the IL-2 group and CIK group (P ＞ 0.05).Conclusion IL-2-transfected CIK cells had stronger killing effects on malignant melanoma.