Objective:To explore the expression and location of TLR5 and NLRC4 on different breast cancer cell lines MDA-MB-231,MCF-7 and MDA-MB-435 and TLR5 activation in breast cancer cell line by recombinant flagellin . Methods:The mRNA level of TLR5 and NLRC4 in MDA-MB-231, MCF-7 and MDA-MB-435 cell were detected with quantitative Real-time PCR and TLR5 expression and location in MDA-MB-231 and MCF-7 cell were detected with Flow cytometry. Induction,expression,purification and i-dentification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97(unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4). 1 μg/ml recombinant flagellin were used to stimulate MCF-7 cell lines,12 h later,the supernate were collected,and ELISA was performed to assess the secretion of IL-8. Results:The mRNA level of TLR5 in MCF-7 cell was 1 700 folds higher than that of MDA-MB-435. TLR5 was expressed in MCF-7 cell surface and ctyosol,while expressed only in cytosol in MDA-MB-231 cell. FliC and FliC-L3A,which were able to activate TLR5 pathway,stimualted MCF-7 cell line to secret IL-8,but FliC△90-97 and FliC△90-97:L3A did not. Conclusion:TLR5 and NLRC4 have been expressed in different breast cancer lines,but there exists difference on the expression level and location of TLR5. Expression level of TLR5 and NLRC4 in MCF-7 cell were higher than other breast cancer lines. TLR5 receptor which is expressed on the surface of breast cancer cell can be activated by flagellin,and these work also provide us experimental basis to further understand the impact of TLR5 activation on breast cancer cell proliferation.

Objective To investigate whether there is prethrombotic state in acute viral encephalitis.Methods Von willebrand factors (VWF),granule membrane protein-140(GMP-140) and D-dimer were measured in 60 children,30 cases of them had mild acute viral encephalitis, and other 30 cases of them had severe acute viral encephalitis.Results The figures mentioned above had marked difference in children with mild acute viral encephalitis during acute and recovery stage compared with controls.Conclusions Testing the expressive rate of GMP-140 on platelet surface,plasma VWF and D-dimer can find the prethrombotic state immediately. GMP-140,VWF and D-dimer in estimating the severe extent of disease and the prognosis play a major role. J Appl Clin Pediatr,2004,19(6):501-503

OBJECTIVETo explore the ultrasound characteristics of intramural pregnancy and evaluate its diagnostic value.

METHODSIntramural pregnancy was diagnosed by endoluminal ultrasonography and confirmed by surgery and pathological examination in 6 patients. The position of the gestational sac, ultrasound findings, distribution of color Doppler flow signals were observed and analyzed together with their surgical and pathological results based on literature review.

RESULTSThere were three types of ultrasound characteristics: gestational cyst type, mass type and uterine rupture type. The gestational cyst type or mass type presented with an empty uterus and a gestational sac or mixed mass was found in the myometrium, which can be separated from the endometrium. The sac or mass did not communicate with the uterine cavity, and abundant blood flow was observed about the sac or in the mass, while massive hemoperitoneum was found in the uterine rupture type.

CONCLUSIONEndoluminal ultrasound is of high value in the diagnosis of intramural pregnancy.

Adult ; Endosonography ; Female ; Humans ; Myometrium ; diagnostic imaging ; Pregnancy ; Pregnancy, Ectopic ; diagnostic imaging ; Ultrasonography, Doppler, Color ; Ultrasonography, Prenatal ; methods

OBJECTIVETo explore the ultrasound characteristics of mucinous cystadenoma of the appendix and evaluate its diagnostic value.

METHODSFourteen cases of mucinous cystadenoma of the appendix were diagnosed by ultrasonic examination and confirmed by surgery and pathology. The ultrasound findings and distribution of color Doppler flow signals were observed and analyzed in comparison with the surgical and pathological results.

RESULTSUltrasound revealed a unilocular or multilocular cystic mass located in the right lower quadrant area with clear boarders, smooth and thick wall, and inconsistent internal echogenicity. The maximal outer diameter of the mass was 2 cm or greater in attachment to the cecum, lacking of periileal inflammatory changes such as in the periileal lymph nodes and echogenic periappendiceal epiploon.

CONCLUSIONUltrasound is of high value in the diagnosis of mucinous cystadenoma of the appendix.

Adult ; Aged ; Appendiceal Neoplasms ; diagnostic imaging ; Cystadenoma, Mucinous ; diagnostic imaging ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Ultrasonography

Adult ; Aged ; Autoimmune Diseases ; diagnosis ; metabolism ; Female ; Humans ; Insulins ; metabolism ; Male ; Middle Aged

Objective:To explore the clinical pathological characteristics and improve the recognition in the diagnosis and treatment of incidental (stage T1a -T1b)prostate cancer.Methods:Seven hundred and seventy-one patients who underwent TURP from May 2004 to September 2013 were analyzed retro-spectively.In our institution,TURP specimens should be totally submitted in an extensive sampling method.The tumor area was outlined by estimation of an experienced genitourinary pathologist and calcu-lated by the image analysis system software (Image J 1.47 h).The tumor area was then multiplied by the thickness of tissue.The total sum of all tumor volume was the estimated tumor volume.The clinical and pathological factors,follow-up results were obtained and we aimed to collect information about the period of watchful waiting (WW),PSA progression status,intervention status during the follow-up,the reason for intervention on WW and the type of intervention.Results:The average age of 771 patients was (71.3 ±5.9)years old,and the average BMI was (23.9 ±3.1)kg/m2 ,preoperative average tPSA was (4.4 ±2.8)μg/L.Eighty-six (11.2%)cases of incidental prostate cancer were detected.The patients in T1a group (77 cases,89.5%)had tumor volumes of (12.3 ±12.6)mm3 ,and the patients in T1b group had tumor volumes of (105.1 ±41.8)mm3 .The range of tumor volume was 0.4 -180.2 mm3 . The volume of all the 86 cases was less than 500 mm3 as the threshold of insignificant cancer.All the pa-tients were managed by WW.The mean follow-up time was 88.9 (27.9 -150.1)months.The Gleason score was <7 in 79 patients,and ≥7 in 7 patients.There was no significant difference in age,preopera-tive tPSA,preoperative PSAD,postoperative tPSA,prostate volume and TURP resection between T1a group and T1b group (P >0.05).Among 84 patients without follow-up losts,PSA progression occurred in 5 patients.One T1a patient underwent radical prostatectomy (RP)as an intervention,and 3 patients underwent hormone therapy.One patient in T1b group underwent radiotherapy for PSA progression and one was treated because of patient preference without evidence of disease progression.There were no pa-tients who died due to prostate cancer.Conclusion:Eighty-six (11.2%)cases of incidental prostate cancer were detected.The tumor volume of all the cases was insignificant cancer.The clinical outcomes of IPCa were satisfactory with the initial treatment of WW in the Chinese population.

Objective To investigate the isolation and culture of the neural stem cells (NSCs) derived from the embryonic mouse spinal cord and striatum, and observe the differences in their proliferation and differentiation characteristics in vitro, thereby providing evidences for identifying more suitable seed cells for repainng spinal cord injuries. Methods The spinal cord and striatum of 14-day-old embryonic mice were dissected for primary NSC culture. After 4 passages, the cells were examined for the expression ofnestin, β-tubulin Ⅲ, glial fibrillary acidic protein (GFAP) and MAG using immunohistochemistry. Results The NSCs derived from the spinal cord and the striatum both possessed in vitro proliferation capacity and expressed nestin antigen. After induced differentiation, the NSCs expressed specific antigens of neurons, astrocytes and oligodendrocytes. Conclusion Multipotent NSCs can be obtained from embryonic mouse spinal cord and striatum through different in vitro culture methods under different conditions. According to the differenees between the two kinds of NSCs in culture situation and differentiation, the striatum derived NSC is suitable for transplantation to repair spinal cord injury.

The prognostic features of T1N0M0 renal cell carcinoma (RCC) in Asian patients have not been well explored in large sample studies. In this study, we retrospectively analyzed the records of 713 patients undergoing nephrectomy for T1N0M0 RCC between 1991 and 2009 in three Asian hospitals. Univariate and multivariate analysis were performed to identify the independent predictive factors for T1N0M0 RCC prognosis among a series of clinicopathological parameters, including age, gender, tumor size, Fuhrman grade, and histological classification. Our results showed that 388 of 713 patients had tumors 4.0 cm or smaller (stage T1a) and 325 of 713 patients had tumors 4.0-7.0 cm in size (stage T1b). Five-year cancer-specific survival (CSS) and recurrence-free survival (RFS) rates for this group of patients were 96.0% and 93.5%, respectively. The patients with T1b RCC had a significantly lower 5-year CSS and RFS rates than did those with T1a RCC (CSS, 93.1% vs. 98.6%, P = 0.026; RFS, 90.0% vs. 96.5%, P < 0.001). Patients with low grade (grades I-II) tumors had a higher 5-year CSS (97.8% vs. 91.2%, P = 0.001) and RFS (95.5% vs. 85.5%, P < 0.001) rate than did those with high grade (grades I-II) tumors. More interestingly, when stratifying patients to T1a and T1b groups, the role of grade in distinguishing prognosis could be only observed in patients with T1b disease. Cox regression showed tumor size and Fuhrman grade were significant in predicting CSS and RFS. Our study suggests that the prognosis of patients with T1N0M0 RCC is excellent, and these results are comparable to previously reported studies in Western patients. Furthermore, our data indicates that patients with T1b disease and high Fuhrman grade have high risk of tumor recurrence and death, thus requiring more frequent follow-up.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asia ; epidemiology ; Carcinoma, Renal Cell ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Nephrectomy ; Proportional Hazards Models ; Retrospective Studies ; Risk Assessment ; Survival Rate ; Tumor Burden ; Young Adult

BACKGROUNDUric acid (UA) is suspected to play a neuro-protective role in Parkinson's disease (PD). This study aimed to evaluate whether the serum UA level was associated with the disease progression of PD in a relatively large population of Chinese patients.

METHODSSerum UA levels were measured from 411 Chinese PD patients and 396 age-matched controls; following the uric acid colorimetric method, the serum creatinine (Scr) levels were also measured to reduce the bias caused by possible differences in renal excretion function. The disease progression was scored by Hoehn and Yahr (H&Y) scales and disease durations; PD group was divided into 3 subgroups according to H&Y scales. Independent-samples t test was performed to analyze the differences between PD group and control group. Multiple analysis of covariance was performed to analyze the differences between PD subgroups. Spearman rank-correlation was performed to evaluate the associations between serum UA or Scr level and disease progression.

RESULTSPD patients were found to have significantly lower levels of serum UA than controls ((243.38 ± 78.91) vs. (282.97 ± 90.80) µmol/L, P < 0.01). As the disease progression, the serum UA levels were gradually reduced. There was a significantly inverse correlation of UA levels with H&Y scales (Rs = -0.429, P < 0.01) and disease duration (Rs = -0.284, P < 0.01) in PD patients of both females and males. No significant difference of the Scr level between PD patients and controls was found ((70.01 ± 14.70) vs. (69.84 ± 16.46) µmol/L), and the Scr level was not involved in disease progression.

CONCLUSIONLower serum UA levels may possess a higher risk of PD, which may be a potential useful biomarker to indicate the progression of PD.

Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; blood ; pathology ; Uric Acid ; blood

OBJECTIVETo clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris.

METHODSINGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA).

RESULTS AND CONCLUSIONThe recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA.

Antigens, Neoplasm ; genetics ; metabolism ; Biomarkers, Tumor ; genetics ; metabolism ; Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Islets of Langerhans ; metabolism ; Lectins, C-Type ; genetics ; metabolism ; Pancreatitis-Associated Proteins ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; metabolism