Objective:To investigate the relationship between the expression mP53 and microvessel density urothelial carcinoma of the bladder and its clinical pathological significance. Methods: Immunohistochemistry(Envision method) was used to detect the expression of mP53 and FⅧ-RAg in 50 tissue specimens of primary BUC and 10 tissue specimens of normal mucosa of bladder .The results were analyzed with the clinic and pathological characters of BUC. Results: The expression of mP53 and MVD had significant differences between the tissues of normal mucosa of the bladder and the tissues of the BUC; but not related to the tumor size. mP53 and MVD value had correlation to different stages (according to UICC-TNM ), different grades (according to WHO), and recurrence(P

Objective The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3.1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.

Objective:To prepare a pig model of obstructive sleep apnea-hypopnea syndrome(OSAHS) and to observe the hemodynamics and pathological characteristics of common carotid artery,so as to lay a foundation for further studying the effect of OSAHS on cardiovascular system.Methods: Twelve male small-type pigs were randomly divided into model group and control group(n=6).Animals in the model group were housed in a negative pressure chamber for 6 months to establish OSAHS model and those in the normal control group were fed routinely.After pigs in the model group presented the symptoms of OSAHS,the changes in hemodynamics of carotid artery were detected with color Doppler ultrasound.The morphological changes of common carotid artery were analyzed under light microscope and electron microscope.Results: Animal model of OSAHS was successfully created.The internal diameter of carotid artery of pigs in the model group was decreased,the intima was increased,and the peak-systolic mean velocity(S) and the resistance index(RI) were both increased compared with those of the control group(P

Objective To observe the effect of amylin on the islet β-cells voltage-gated L-calcium channels in rats. Method Patch clamp technique was employed in the observation of the features and changes of electric current of islet β-cells voltage-gated L-calcium channels before and after using amylin. Results In the glucose environment of 5. 5 mmoL/L, the electric current of rat islet β-cells voltage-gated L-calcium channels was activated at-40 mV and reached the peak at about +20 mV, with a peak value of about-120 pA and the insulin secretion level was(0. 76±0. 12) μg/L. Under the stimulation of glucose of 16. 7 mmol/L, the peak current voltage moved to the left and increased up to-140 pA and the level of insulin secretion measured (1.78±0. 13) μg/L. Hatch islet β-cells in amylin at the concentrations of 0. 5, 1.0, 5.0 and 10.0 μmol/L, respectively. It was observed that in the 0. 5 μmol/L and 1.0 μmol/L groups,there was no remarkable change in the peak potential activation voltage, current, and insulin secretion volume in comparison with the control group. However, in the environment of 5.5 mmol/L glucose, the increase of activation voltage of the 5.0 and 10.0 μmol/L groups was-30 mV, with the peak current reduced to approximately-80 pA and-60 pA and the insulin secretion decreased to (0. 49±0. 11) μg/L and (0. 36±0. 12) μg/L respectively. Under the concentration of 16. 7 mmol/L glucose, the activation voltage increased from-40 mV up to-30 mV and the peak current reduced to-80 pA and-40 pA. In the meantime, the insulin secretion decreased respectively to (1.20±0. 13) μg/L and (0. 89±0. 14) μg/L, which is of significance. Conclusion The secretion of insulin is synchronized with the opening of the islet β-cells voltage-gated L-calcium channels at the stimulation of glucose. The amylin inhibition of the insulin secretion is also synchronized with the opening of islet β-cells voltage-gated L-calcium channels and it's in a positive concentration-dependent manner.

Objective:To establish a rapid detection method for human astrovirus based on TaqMan-probe real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR).Methods:According to the conservative sequence of human astrovirus ORF1 b gene, we designed the amplification primers and specific fluorescent probe to establish the human astrovirus TaqMan real-time fluorescent quantitative RT-PCR rapid detection method. The specificity, sensitivity and stability of the method were evaluated. We also used this method to detect human astrovirus in clinical samples. Results:The established human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method has good specificity and repeatability for human astrovirus, and the sensitivity can reach 10 2 copies/μl. After testing the clinical samples, the detection rate of human astrovirus by our method was 100%. Conclusions:The human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method established in this study is simple, rapid, sensitive, specific and stable. It can be used for clinical human astrovirus detection and epidemiological investigation.

AIM: To investigate whether rs7903146 polymorphisms in transcription factor 7-like 2 (TCF7L2) gene are associated with susceptibility to type 2 diabetes mellitus (T2DM) in Chinese Uygur population.METHODS: In this case-control study, 935 cases of T2DM patients were recruited in T2DM group, and 971 healthy examination individuals were selected as normal control.The TCF7L2 gene polymorphism was detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrum.RESULTS: Significant differences in the frequencies of CC, CT and TT genotypes and the frequencies of C and T alleles on TCF7L2 rs7903146 were found between T2DM group and control group(P<0.05).As compared with C allele, the patients with T allele had a significantly higher risk of T2DM with OR of 1.190 (95% CI: 1.034~1.371).As compared with CC genotype, the patients with CT genotype had a significantly higher risk of T2DM with OR of 1.374 (95% CI: 1.122~1.683), and the patients with CT+TT genotype had a significantly higher risk of T2DM with OR of 1.307 (95% CI: 1.090~1.567).The levels of fasting plasma glucose, serum creatinine and blood urea nitrogen were higher in all participants with CT+TT genotype of rs7903146 than those with CC genotype, which showed a significant difference (P<0.05).CONCLUSION: The polymorphisms of rs7903146 in TCF7L2 gene may be associated with T2DM in Uygur population from Xinjiang region.The T allele and CT genotype of rs7903146 are the risk factors for T2DM.

Objective To investigate the correlation between TCF7L2 gene rs3814570 polymorphisms with type 2 diabetes mellitus(T2DM) in Uygur population of Xinjiang area.Methods By adopting the case-control study design,949 cases of T2DM were recruited as the observation group and 963 individuals Undergoing healthy physical examination were selected as the control group.The TCF7L2 gene polymorphism was detected by matrix-assisted laser desorption/ionization-time of flight(MALDI-TOF).Results The statistical differences in frequencies of CC,CT and TT genotypes and the C and T allele frequencies on TCF7L2 rs3814570 were found between the T2DM group and control group(P＜0.05).The risk of suffering from T2DM in the carriers of CT genotype was 0.331 times of that in the carriers of CC genotype(OR =0.331,95 ％ CI:0.166-0.661,P =0.002),the risk of suffering from T2DM in the carriers of TT genotype was 0.539 times of that in the carriers of CC genotype(OR=0.539,95％CI:0.348-0.834,P=0.005),and the risk of suffering from T2DM in the carriers of T allele was 0.501 times of that in the carriers of C allele(OR=0.501,95 ％ CI:0.377-0.664,P＜ 0.01).Among all subjects,the FPG level of the CT + TT genotype group on TCF7L2 gene rs3814570 locus was significantly lower than that of the CC genotype group(P＜0.05).Conclusion The rs3814570 locus in TCF7L2 gene may be associated with T2DM occurrence in Uygur population of Xinjiang area,the T allele and TT genotype might be protective factors of T2DM.

Objective To explore the relationship between traditional Chinese medicine (TCM) syndromes of state of evil domination and clinical and laboratory indicators of IgA nephropathy. Methods A prospective study was used to collect data on clinical and laboratory examination of IgA nephropathy in multi clinical centers. Patients’ TCM syndrome types were determined according to the national diagnostic criteria at the same time. Totally 266 patients with IgA nephropathy were included in the study to establish a database for analyzing the relationship between TCM syndrome of state of evil domination and clinical and laboratory indicators. Results In the relationship between syndromes of state of evil domination distribution and clinical subtypes, patients of wind-heat syndrome had more macroscopic hematuria;patients of phlegm-damp syndrome had more nephrotic syndrome;patients of damp-heat syndrome had more chronic nephritis type Ⅰ;patients of blood stasis syndrome had more chronic nephritis type Ⅱ. In the relationship between syndrome of state of evil domination distribution and clinical manifestation, the incidence of hypertension was higher in patients of blood stasis syndrome than in other three types. It was more serious for hematuresis in the patients of wind-heat syndrome. For patients of phlegm-damp syndrome, the incidence of heave proteinuria was highest. In the relationship between syndrome of state of evil domination distribution and laboratory examination, 24-hour urinary protein quantification was higher than in patients of wind-heat and damp-heat syndrome, but the level of blood albumin was lowest. For patients of blood stasis syndrome, serum creatinine level was significantly higher than in other three types;the level of eGFR was just the opposite. The levels of blood cholesterol and triglyceride in patients of phlegm-damp syndrome were higher than in other three types. The activated partial thromboplastin time (APTT) levels in patients of blood stasis and phlegm-damp syndrome were lower than in other patients, but the FIB level was the exact opposite of APTT. In the relationship between syndrome of state of evil domination distribution and the stages of chronic kidney disease (CKD), patients of wind-heat syndrome were more in the first stage of CKD;patients of blood stasis syndrome were more in the third stage of CKD. Conclusion There is relative correlation between TCM syndromes of state of evil domination and clinical and laboratory indicators in IgA nephropathy, which would provide some reference to narrow the gap in the information of the four methods of TCM with clinical and laboratory indicators to enhance accurate diagnosis of TCM syndrome.

Objective To investigate the expression level and clinical significance of WT1 gene in acute luekemia (AL) patients.Methods WT1 gene level was detected by real time quantitative-polymerase chain reaction in acute myelogenous leukemia (AML) and in acute lymphocytic leukemia (ALL) patients.Then the expression levels of WT1 gene in different subtypes of AML were compared,and the correlation between gene expression and disease courses and prognosis were observed.Moreover,the relationship between disease courses and WT1 expression in patiens after receiving haemopoietic stem cell transplantation were analyzed.Results Among 66 cases,WT1 expression positive rate was 87.5 ％ (14/16) in AML and 76.0 ％ (38/50) in ALL.In AML,the expression level in M3 showed the lowest than that in any other subtypes (compared with M1,M2,M4,M5,P value was 0.040,0.007,0.006 and 0.01,respectively).The expression level of WT1 was closely correlated with leukemia disease courses.The expression level in complete remission (CR) group showed a significant lower expression level than that in non-remission group (P =0.018) and relapse group (P =0.003),and the re-increase of WT1 expression level could predict relapse as early as 1.5 months.Moreover,WT1 expression also showed an close relationship with prognosis of patients receiving haemopoietic stem cell transplantation.Patients whose WT1 was undetectable had a better prognosis than those with persistent expression,and increase again after becoming undetectable.Conclusion WT1 has a high expression level in AL,which can represent minimal residual disease.The expression level in M3 was lowest than that in different AML subtypes,and its expression level has a close correlation with clinical disease course and prognosis of AL.