Objective To analyze tissue vascular endothelial growth factor receptor 1 (VEGFR-1)and VEGFR-2 and their soluble form (sVEFGR-1 and sVEGFR-2) in the plasma in patients with preeclampsia,and to explore its clinical significance.Methods sVEGFR-1 and sVEGFR-2 expressions in plasma of normal pregnancy women and preeclampsia patients were detected by enzyme-linked immunosorbnent assay (ELISA) ; VEGFR-1,VEGFR-2,sVEGFR-1 and sVEGFR-2 mRNA expressions of normal pregnancy women and preeclampsia patients in placenta were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results ELISA results showed that the level of sVEGFR-1 in plasma of normal control group before and after delivery was (12.33 ± 1.52) ng/ml and (4.55 ± 0.31) ng/ml,respectively,while that of preeclampsia group was (77.25 ± 9.47) ng/ml and (8.13 ± 0.74) ng/ml,respectively; the differences between before and after delivery in the two groups were of statistical significance.The level of sVEGFR 2 in plasma of normal control group before and after delivery was (8.74 ± 1.24) ng/ml and (6.43± 0.55) ng/ml,respectively,while that of preeclampsia group was (5.69 ± 0.75) ng/ml and (4.96 ±0.67) ng/ml,respectively.RT-PCR results were consistent with ELISA results.Conclusions Rapid decrease of plasma sVEGFR-1 and continuously low-level expression of plasma sVEGFR-2 indicated that VEGFR-1 might be closely related to preeclampsia,and decrease of plasma sVEGFR-2 in preeclampsia women might be taken as a marker of endothelial cell function disorder.

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.

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.

Objective To explore the mechanism of latent human immunodeficiency ciency virus type 1 (HIV-1) infection is unclear, especially in dendritic cells (DC).We hypothesized that DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) binds with HIV-1 may activate HIV-1 provirus.Methods We generated a model by transfecting 293T cells with a DC-SIGN expression plasmid and a HIV-1 5'long terminal repeat (LTR) reporter plasmid, and then stimulated the 293T cells with HIV-1 gp120 protein, wild-type HIV-1 and VSV-G-pNL4.3 pseudotype virus ( without gp120 protein).CEM-Bru cells were transfected with the DC-SIGN expression plasmid and stimulated by HIV-1 gp120 protein.Then HIV-1 replication was detected.The involvement of the ERK, p38 and NF-κB pathways signaling in this response were determined by inhibiting the pathways specifically and detecting the phosphorylation of the signaling kinase.Results The HIV-1 5'LTR was reactivated by HIV-1 gp120 in DC-SIGN-expressing 293T cells.After HIV-1 gp120 protein stimulation of the mold of CEM-Bru cells, the increasing expression of HIV-1 Tat mRNA and HIV-1 p24,which implies early and late HIV-1 provirus replication was reactivated by the HIV-1 gp120/DC-SIGN stimulation.HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.Conclusion HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.

Objective To observe the effects of modified Yigong Powder (MYP) combined with chemotherapy on transplanted hepatocarcinoma of mice and to study its mechanisms. Methods Hepatocarcinoma 22 mouse model was established and then was used to observe the attenuating and synergic effects of MYP when applied together with 5-fluorouracil (5-FU). After 8 days of treatment,the tumor-inhibiting rate,activity of natural killer (NK)cells and interleukin-2 (IL-2) and small intestinal malondialdehyde (MDA) content were examined. Results MYP combined with 5-FU could increased the tumor-inhibiting rate to some extent and improve the immune function by increasing immune organ weight and increasing the activity of NK cells and IL-2. MYP combined with 5-FU could also reduce the 5-FU-induced intestinal injuries by relieving the damage of free radicals and inhibiting the lipid peroxidation and a good prognosis was expected in tumor-bearing animals treated with chemotherapy. Conclusion MYP exerts an attenuating and synergic effect when used together with 5-FU in treating tumor-bearing mice and its mechamism may be related to the improvement of immune function and reduction of intestinal injuries.

This study provides the candidate sequences in the identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding. We amplified and sequenced the region psbA-trnH, with the data of 284 sequences from GenBank, the differential intra- and inter-specific divergences, genetic distance, barcoding gap were used to evaluate five barcodes, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The results showed that psbA-trnH barcodes performed high identification efficiency and inter-specific divergences among the five different DNA barcodes. Analysis of the barcoding gap and NJ tree showed psbA-trnH was superior to other barcodes. Based on the identification and PCR amplification efficiency, psbA-trnH can be the ideal barcode to identify Radix et Rhizoma Clematidis and its adulterants accurately.

To establish a new method for identifying genus of Lilium by DNA barcoding technology, ITS, ITS2, psbA-trnH, matK and rbcL sequences were analyzed in term of variation of inter- and intra-species, barcoding gap, neighbor-joining tree to distinguish genus of Lilium based on 978 sequences from experimental and GenBank database, and identification efficiency was evaluated by Nearest distance and BLAST1 methods. The results showed that DNA barcoding could identify different species in genus of Lilium. ITS sequence performed higher identification efficiency, and had significant difference between intra- and inter-species. And NJ tree could also divide species into different clades. Results indicate that DNA barcoding can identify genus of Lilium accurately. ITS sequence can be the optimal barcode to identify species of Lilium.

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 assess the noninvasive index for diagnosing the degree of esophageal varices (EV) in patients with hepatitis B virus (HBV)-related cirrhosis. Methods The clinical data of 112 patients with HBV-related cirrhosis were studied retrospectively. The patients were divided into non-EV (NEV) group (30 cases) and EV group (82 cases) according to the results of gastroscopy. In EV group, there were mild varices in 21 cases (mild EV group), moderate varices in 47 cases (moderate EV group) and severe varices in 14 cases (severe EV group). The age, gender, platelets, glutamyl transpeptidase, prothrombin time, albumin, bilirubin, portal vein diameter, spleen vein diameter and thickness of spleen were compared, and the relationship was analyzed between each index and EV. Results There were no statistical differences in gender, age, albumin, bilirubin, prothrombin time and glutamyl transpeptidase between NEV group and EV group (P>0.05). The portal vein diameter, spleen vein diameter and thickness of spleen in EV group were significantly higher than those in NEV group:(14.1 ± 3.1) mm vs. (10.6 ± 2.3) mm, (8.9 ± 2.1) mm vs. (7.6 ± 1.6) mm and (4.8 ± 0.9) mm vs. (3.8 ± 1.0) mm, the platelets in EV group was significantly lower than that in NEV group:(86.8 ± 20.2) × 109/L vs. (163.5 ± 18.1) × 109/L, and there were statistical differences (P<0.01 or<0.05). The portal vein diameter, spleen vein diameter and thickness of spleen in moderate EV group and severe EV group were significantly higher than those that in NEV group and mild EV group: (13.5 ± 2.1) and (14.8 ± 3.6) mm vs. (10.6 ± 2.3) and (11.2 ± 3.1) mm, (8.3 ± 2.1) and (9.1 ± 1.1) mm vs. (7.6 ± 1.6) and (8.1 ± 1.9) mm, (4.7 ± 1.1) and (4.9 ± 0.9) mm vs. (3.8 ± 1.0) and (4.1 ± 1.2) mm, the platelet levels were significantly lower than those in NEV group and mild EV group: (72.8 ± 11.6) × 109/L and (63.8 ± 15.6) × 109/L vs. (163.5 ± 18.1) × 109/L and (100.2 ± 10.3) × 109/L, and there were statistical differences (P<0.05). The area under curve of response operating characteristic for predicting the presence of moderate and severe EV with portal vein diameter and platelets were 0.719 and 0.735, and the cut off value were 14 mm and 69 × 109/L. Conclusions The portal vein diameter and platelets can predict the presence of moderate and severe EV in patients with HBV-related cirrhosis.

10.3969/j.issn.2095-4344.2013.23.004