Objective:To explore the correlation with neonatal congenital cytomegalovirus infection,maternal primary infection and secondary infection.Methods: 48 neonates with congenital CMV infection were assigned to infection group with their mothers.And the other 30 couples without congenital CMV infection were assigned to negative group with their mothers.The level of CMV-IgM/IgG and affinity of CMV-IgG in peripheral blood were tested by CLIA,and CMV-DNA in mother′s milk,peripheral blood and urine of the newborn was tested by fluorescent quantitation PCR.We also analyzed the differences of the test results between the two groups and performed a retrospective analysis to compare the levels of CMV-IgG of the mother with early pregnancy with the result of this test.Results: In the infection group,the level of CMV-IgG in peripheral blood and CMV-DNA in milk was significantly higher than those in the negative group,the difference was statistically significant(P<0.01).Ratio of CMV-IgG antibody in newborn babies and their mothers in the infection group was lower than the other group,the difference was statistically significant(P<0.01).There was negative correlation of IgG level between the newborn babies and their mothers in the infection group,the difference was statistically significant(P<0.01);While in the positive correlation in the negative group,the difference was statistically significant(P<0.01).The CMV-IgG concentrations of the mothers with early pregnancy was significantly lower than that of this infection group,the difference was statistically significant(P<0.01);but in the negative group,there was no significant difference between the CMV-IgG level of the mothers with early pregnancy and the results of this test(P>0.05).Conclusion: It is a high-risk factor for neonatal congenital cytomegalovirus infection that CMV-IgG level of the pregnant women is promted by the reactivation or reinfection of cytomegalovirus.It is important to monitor CMV-IgM/IgG during pregnancy.

Objective To investigate the relationship between EphA7 protein and carcinogenesis and development of pancreatic cancer by detecting the expression of EphA7 protein in pancreatic cancer tissues. Methods The expression of EphA7 in 10 cases of normal pancreatic tissue, 51 cases of pancreatic cancer and its adjacent tissues were detected with immunohistechemieal methods and the relationship between EphA7 and pathologic features of pancreatic cancer were analyzed. Results The rate of expression of EphA7 protein in normal pancreatic tissue was 10% (1/10), 47.1% (24/51) in adjacent pancreatic cancer tissues, 94.1% (48/51) in pancreatic cancer tissues. There were significant difference among the three groups (P <0.05). There was no significant correlation between the expression of EphA7 protein and the age, sex, tumor location, tumor size in patients with pancreatic cancer (P > 0.05). However there was significant correlation between the expression of EphA7 protein and the degree of differentiation, clinical staging, lymph node metastasis, or distant metastasis (P < 0.05). Conclusions The abnormally high expression of EphA7 may be relevant with the occurrence and development of the pancreatic cancer.

Objective To establish a fast and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation.Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations` distribution,each tube of cell fraction was then partially transfered to the 96 plate well,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy.Results Three types of spermatogenic cells can be identified quickly and accurately by it`s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection.Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation is successfully developed.

Objective To investigate the effect of retinoblastoma binding protein 4 (RBBP4)in Sp1 -mediated HIV long terminal repeat(LTR)transcription.Methods RBBP4 expression vector and Sp1 expression vector were respectively co-transfected into 293 T cells with HIV promoter pHIV-LTR-Luc or Sp1 site mutated pHIV-LTR-sp1 -mut by liposome transfection,and the transfected cells were examined by dual luciferase reporter assay system.The effect of RBBP4 on the binding of Sp1 to LTR was further studied by chromatin immunoprecipitation (ChIP)and electrophoretic mobility shift assay (EMSA).Results The relative firefly luciferase activity activated by Sp1 was decreased from 62.5 to 16 at the dose of 500 ng of RBBP4 expression vector (t =14.52,P <0.01 ).When the Sp1 binding sites were mutated,the effects of 100,300 or 500 ng of RBBP4 expression vector on the firefly luciferase activity of HIV LTR were not statistically significance (t =1 .897,2.357 and 3.162,all P <0.05).ChIP results showed that when the binding of RBBP4 on HIV LTR increased,the binding of Sp1 on HIV LTR increased significantly (t =11 .93,P <0.01 ),while the reduced binding of RBBP4 on HIV LTR significantly attenuated the binding of Sp1 onto LTR(t =11 .38,P <0.01 ).The effect of RBBP4 on the binding of Sp1 to DNA in ChIP assays was further verified by EMSA assays.Conclusion RBBP4 can inhibit the Sp1 -mediated HIV LTR transcription in 293 T cells.

The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen activation or NF-kappaB activation in rats was studied in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-kappaB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10(-3) mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P < 0.01) following the treatment with a high concentration of carbachol (10(-3) mol/L) in vitro. The addition of 10(-2) mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10(-3) mol/L) in vitro (P > 0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-kappaB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.
Carbachol/*pharmacology ; Cholinergic Agonists/pharmacology ; NF-kappa B/*metabolism ; Pancreas/metabolism ; Pancreas/*pathology ; Rats, Wistar ; Receptor, Muscarinic M3/agonists ; Trypsinogen/*metabolism

We aimed to examine the effect of pioglitazone on transdifferentiation of preosteoblasts from rat bone marrow mesenchymal stem cells (BMSCs) into adipocytes and investigate its effect on bone metabolism. BMSCs were harvested from the femurs and tibias of a rat, then separated, purified, proliferated for 3 generations and differentiated into preosteoblasts for 5 days and 14 days respectively in the presence of osteogenic medium. Thereafter, the preosteoblasts were cultured for 21 days in the presence of adipogenic medium with and without pioglitazone (1 μg/mL). Partially-differentiated osteoblasts were identified by mineralized nodules with Alizarin red S staining. Transdifferentiated adipocytes were identified by Oil Red O staining. Reverse transcription PCR (RT-PCR) was performed to assay the expression levels of osteogenic markers Runx2 and ALP, and an adipogenic marker PPARγ. Those cells cultured for 5 days did not show mineralized nodules as detected by staining of Alizarin red S, while those cultured for 14 days showed dispersed mineralized centers in the form of brown spots, although without obvious red mineralized nodules. After adipogenic transdifferentiation for 21 days, adipose-drops were found in cells of 5CG and 5EG earlier than those of 14CG and 14EG, and the former showed much more adipocytes separately as detected by Oil Red O staining. Whatever the time was 5 days or 14 days of BMSCs osteogenic differentiation, the cells cultured with pioglitazone showed much more adipocytes than those without pioglitazone. Our experiment showed that the less time it took for BMSCs osteogenic differentiation, a stronger ability remained for BMSCs to transdifferentiate into adipocytes. The mRNA expression levels of Runx2 and ALP were decreased by 1.79 and 1.90 times respectively in 5EG (P< 0.05) as compared with 5CG, and that of PPARγ was increased by 1.31 times in 5EG (P<0.05) as compared with 5CG. The mRNA expression levels of Runx2 and ALP were decreased by 1.45 and 1.54 times respectively in 14EG (P<0.05) as compared with 14CG, and that of PPARγ was increased by 1.39 times in 14EG (P<0.05) as compared with 14CG. It was concluded that pioglitazone stimulated the transdifferentiation of BMSCs into adipocytes. These observations provided a potential mechanism of imbalance in thiazolidinedione induced bone metabolism.

The aim of this study was to evaluate the seed quality of C.deserticola and establish quality grading rules of seeds by detecting the impacts of different processing methods on the contents of the effective components of C.deserticola for optimizing the suitable processing method.The seed quality was judged by thousand-kernel weight,empty embryo rate and water content.The samples were preliminary processed by freeze-drying,natural drying and hot air circulation drying respectively,and the content of phenylethanoid glycosides was determined by HPLC-UV.The seed quality classification standard of C.deserticola was established,and the seeds were divided into three grades based on the standard.It was found that freeze-drying method was optimum,featuring less effective component loss,beautiful appearance of herbal piece,crisp texture and fast drying.In conclusion,this study laid a foundation for the quality control of the seeds of C.deserticola with the provision of scientific evidence for the initial processing of the fresh product.

Objective To compare the safety and effectiveness of power peripherally inserted central catheters (Power PICC) and double cavity central venous catheter (CVC) application in intensive care unit (ICU). Methods 458 cases were reviewed during January to September in 2014 and divided into two groups: Power PICC group (245 cases) and CVC group (213 cases) , and average retention time, successful rate of inserting catheter and the incidence of complications were compared. Results The average retention time of Power PICC group was (21.6±5.8) days which was longer than (13.1±3.4) days of CVC group (t=2.234, P <0.05). No statistics difference of successful rate between two groups (P>0.05). No significant difference for the total incidence of complications between two groups as 14.69% (36/245) and 19.72%(42/213)(P>0.05). No significant difference for the total incidence of complications in the operation time between two groups as 5.31% (13/245) and 4.23% (9/213)(P>0.05). But rate of catheter malposition for Power PICC group [ 2.86% (7/245) ] was higher than CVC group 0 (X2=4.428, P <0.05). Rate of the total incidence of complications in the retention time Power PICC group [ 9.39%(23/245) ] was lower than CVC group [ 15.96%(33/213)(P<0.05). And rate of catheter related blood stream infection of CVC group [3.29%(7/213)] was much more higher than Power PICC group (0)(X2=6.139,P<0.05). Conclusions Power PICC and CVC are both applicable for ICU, and Power PICC has more advantage regarding safety and effectiveness than CVC and can be one replacement for CVC.

Objective:To investigate the diagnostic and terapeutical value of FPG,GA,HbA1c and GA/HbA1c ratio in T1DM/T2DM.Methods:The study was made by case-control method.In our study,30 healthy subjects were selected from health physical ex-amination as control group while 160 diabetics were selected as case group,in which there are 76 TIDM and 84 T2DM.Analyzing the difference of relevance of FPG,GA and HbA1c,the difference of GA/HbA1c and threshold of the case and the control,and this analysis was also used between the T1DM and the T2DM.The data was managed by independent-sample t test,ROCK and Pearson correlation test of SPSS.Results:The results of FPG,GA ,HbA1c and GA/HbA1c ratio of T1DM and T2DM were significantly higher than those in the control group(P<0.01).And they were higher in the T1DM than in the T2DM(P<0.01);in the T1DM group,GA was strongly positive correlative with HbA1c(P<0.01),FPG was weakly positive correlative with GA(P>0.05),and weakly negative correlative with HbA1c(P>0.05);in T2DM group,there were positive correlation among FPG,GA and HbA1c(P<0.05),the degree of correlation ranked as HbA1c/GA>FPG/GA>FPG/HbA1c;analyzing the ROC of measures in T1DM group,the sensitivity and specificity were re-spectively 86.8% and 100% for diagnosing DM when FPG threshold was set on 5.86 mmol/L ( AUC=0.922 ) ( P<0.05 ).The sensitivity and specificity were both 100% for detecting DM when GA threshold was set on 15.5%( AUC=1 ) ( P<0.05 ).The sensitivity and specificity were respectively 98.7%and 100%for diagnosing DM when HbA1c threshold was set to 6.10%( AUC=1) ( P<0.05).The sensitivity and specificity were respectively 93.4% and 100% for diagnosing DM when the threshold of GA/HbA1c was set on 2.95 ( AUC=0.992 ) ( P<0.05 );analyzing the ROC of measures in T2DM group, the sensitivity and specificity were respectively 91.7% and 100% for diagnosing DM when FPG threshold was set on 5.94 mmol/L ( AUC=0.941 ) ( P<0.05 ).The sensitivity and specificity were respectively 85.7%and 100%for diagnosing DM when GA threshold was set on 15.5%( AUC=0.977) ( P<0.05).The sensitivity and specificity were respectively 97.6% and 100% for diagnosing DM when HbA1c threshold was set on 5.95%(AUC=0.991)(P<0.05).The ratio of GA/HbA1c had no diagnostic cutoff point,AUC was 0.644(P>0.05).Conclusion:FPG,GA, HbA1c and GA/HbA1c ratio are of high value in monitoring of blood glucose, diagnosis and typing in T1DM and T2DM.There are missed diagnosis when we diagnose T1DM and T2DM by the upper limit of reagent instruestion of FPG,GA,HbA1c.It is more important for a person with T1DM to monitor FPG than others.

Objective To explore the value of peripheral blood samples in KRAS mutation testing of colorectal cancer patients and the correlation between the number of circulating tumor cells and KRAS mutation testing.Methods We detected KRAS mutation using amplification refractory mutation system PCR method in paraffin embedded tissues and matched peripheral blood samples obtained from 112 colorectal cancer patients and 10 proctitic peripheral blood samples in Affiliated Hospital of Jiangnan University between 2013 and 2014.Meanwhile,immunofluorescence in situ hybridization method was used to count the circulating tumor cells in peripheral blood samples and proctitic control samples.Results Among the 112 colorectal cancer samples tested,25 cases of peripheral blood samples found KRAS mutation (41.1％) and which was 46 in formalin fixed paraffin embedded tissues testing (22.3 ％),with a significant difference (x2 =40.12,P ＜ 0.001).One case with KRAS wild type in formalin fixed paraffin embedded tissues was mutation type in peripheral samples.In another case,mutation site was different in different kinds of samples.The sensibility of KRAS mutation testing was 73.3％,41.9％ and 16.7％ when the number of circulating tumor cells was more than 15,5 to 15,and 1 to 5,respectively,with significant differences (x2 =23.70,P ＜ 0.001).No KRAS mutation and no circulating tumor cells were found in 10 proctitic control samples.Conclusion We find high specificity in KRAS mutation testing of peripheral blood samples.but the accurate rate is not satisfying.KRAS mutation testing in peripheral blood samples may be an optional choice to test KRAS mutations for colorectal cancer patients who were not subjected to surgery.The sensibility of KRAS mutation testing in peripheral blood samples has a corretion with the number of circulating tumor cells.