Objective To investigate the relationship among the activity of protein kinase C (PKC) in platelets of maternal vein and umbilical blood , the pathophysiological changes of pregnancy induced hypertension (PIH) and fetal growth restriction (FGR) in PIH patients. Methods Activities of PKC in membrane and plasma of platelets from maternal vein and umbilical blood taken from 35 PIH patients and 20 normal pregnant women were measured with substrate phosphorylation method. Results No difference was shown in the PKC activities between the mild PIH patients and normal pregnant women in both maternal and cord blood.The PKC activities in moderate and severe PIH patients were significantly higher than those of the normal pregnant group.In normal pregnant women, the PKC activity in membrane and plasm of the platelets had no significant difference. In the moderate and severe PIH group, PKC activity in membrane was far more higher than the plasm 46?6 vs 37?4 pmol/(min?mg protein), P

Objective To investigate the role of hypoxia inducible factor(HIF)-prolyl hydroxylase 1 (HPHl)and factor inhibiting HIF-1(FIH-1)in placentas in the pathogenesis and development of severe pre-eclampsia.Methods RT-PCR and western blot analyses were used to detect the HPH1 and FIH-1expression levels in placentas of 34 patients with severe pre-eclampsia and 24 cases of term pregnancy (normal pregnancy group)and their correlations with symptoms were analyzed.Results (1)The HPHI mRNA and protein expression levels in placentas of severe pre-eclampsia group were 0.40±0.04 and 59.5±3.4 separately,significantly lower than those of normal pregnancy group,0.84±0.12 and 71.6±1.7(P<0.01).The FIH-1 mRNA and protein expression levels in placentas of severe pre-eclampsia group wereQ 31 ±0.05 and 45.6±2.4 separately,significantly lower than those of normal pregnancy group,0.43±0.04 and 54.9±2.1(P<0.01).(2)The mRNA and protein expression levels of HPH1 and FIH-1 in severe pre-eclampsia group were all negatively correlated with mean arterial pressure(MAP)[the Spearman correlation coefficient was-0.854(P<0.01)],urinary protein per 24 hours[the Spearman correlation coefficient was-0.936(P<0.01)1 and the occurrence of fundus oculi artery spasm[the Spearman correlation coefficient was-0.854(P<0.01)].(3)rrhe expression of HPHl mRNA in placentas of all the 58 cases WBB 0.58±0.27.higher than the expression of FIH-1 mRNA,which was 0.39±0.10.There was a positive correlation between them.The pearson correlation coefficient was 0.686(P<0.01).The expression of HPH1 protein in placentas of all the 58 cases was 64.5±6.7,higher than the expression of FIH-1,which was 49.4±5.2.There was a positive correlation between them.The Pearson correlation coefficient was 0.947(P<0.01).Conclusion The expression imbalance of HPH1 and FIH-1in palcenta may play an important role in the pathogenesis and development of severe pre-eclampsia through inhibiting HIF-1a.

Objective To evaluate different expressions of high mobility group box 1(HMGB1)and receptor for advanced glycation end products(RAGE)in placentas and their relationship with preeclampsia.Methods Fifteen early-onset pre-eclaraptic women(early-onset pre-eclampsia group),22 late-onset pre-eclamptic women(late-onset pre-eclampsia group)and 12 normotensive women(control group)in the third trimester were recruited at the Shengjing Hospital of China Medical University from March 2006 to March 2007.The localization and levels of HMGB1 and RAGE in placentas of the three groups were detected by the strept avidin biotin-peroxidose method.Results (1)Immunoreactivities to HMGB1:positive immnnostaining for HMGB1 was observed in trophoblast,macrophages,decidual cells,vascular muscle cells,endothelial cells and placental mesenchymal cells in the placentas from the pre-eclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies;the staining was observed within both the nuclei and the cytoplasm,mainly in the cytoplasm.The cytotrophoblast,especially the nuclei was extensively positive for HMGB1 in early-onset pre-eclampsia. (2)Immunoreactivities to RAGE:positive immunostaining for HMGB1 was observed in syncytiotrophoblast,macrophages and endothelial cells in the placentas from the preeclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies:the staining was in the cytoplasm and(or)cell membrane.The trophoblast was extensively positive for RAGE in early-onset pre-eclampsia.(3)Positive rate of HMGB1 expression:the expression of HMGB1 in early-onset group(73%,11/15)and late-onset group(64%,14/22)was significantly higher than that in normal group(17%,2/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).(4)Positive rate of RAGE expression:the expression of RAGE in early-onset group(80%,12/15)and late-onset group (82%,18/22)was significantly higher than that in normal group(25%,3/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).Conclusions The increased expression of HMGB1 and RACE in the placenta may play an important role in the pathogenesis of pre-eclampsis.The different locations may be associated with the occurrence of different onset types of pre-eclampsia.

Objective:To investigate the expression of nicotinamide adenine dinucleotide (NAD +) digestive enzyme CD38 in normal and endotoxin-tolerant human monocyte THP-1 cell lines treated with lipopolysaccharide (LPS). Methods:(1) Normal THP-1 cells: The experiment and control group were treated with 100 ng/ml LPS for 1, 3, 6, 12 and 24 h or phosphate buffer for 24 h, respectively. Quantitative polymerase chain reaction (PCR) and Western blot were used to measure the expression of interleukin-6 (IL-6) and tumor necrosis factor (TNF) mRNA, CD38 mRNA and protein. (2) The induced endotoxin-tolerant THP-1 cells: ①The model of endotoxin-tolerant cells was established firstly by treating the THP-1 cells with 100 ng/ml LPS for 24 h. THP-1 cells treated with phosphate buffer were set as blank group. After further stimulating the two groups with LPS (100 ng/ml) for 3 h, mRNA levels of IL-6 and TNF were measured by quantitative PCR to determine whether the modeling was successful or not. ②In addition, the expression of CD38 mRNA were detected by quantitative PCR before and 12 h after LPS stimulation, and the expression of CD38 protein of these two groups were detected by western blotting before and 1, 6 h after LPS stimulation. Two independent samples t-test and repeated measurement analysis of variance were used for statistical analysis. Results:(1) In normal THP-1 cells, the mRNA expression levels of IL-6 and TNF in the LPS-stimulated cells were significantly higher than those of the control at all time points. And a higher expression level of CD38 mRNA and protein was observed in LPS-stimulated cells from 3 to 24 h compared with the control (mRNA at 3 h: 2.27±0.03 vs 1.00±0.18; protein at 3 h: 1.47±0.14 vs 1.00±0.16, both P<0.05). (2) Endotoxin-tolerant THP-1 cells: ①IL-6 and TNF mRNA levels in the model group were significantly lower than those in the blank group (both P<0.05), indicating that the endotoxin-tolerant THP-1 cell model was established successfully. ②Compared with the same points in the blank group, CD38 mRNA expression was upregulated in the model group before stimulating by LPS (14.18±1.19 vs 1.00±0.13, t=19.007) and 12 h after LPS stimulation (28.33±3.98 vs 7.61±0.88, t=8.803). Moreover, CD38 protein levels before stimulating by LPS (1.54±0.06 vs 1.00±0.10, t=7.796) and 1 h (1.59±0.09 vs 1.07±0.17, t=4.721), 6 h after LPS stimulation (2.48±0.09 vs 1.43±0.12, t=12.233) in the model group were all higher than those in the control group (all P<0.05). The intra-group comparison showed that in the model group the levels of CD38 mRNA at 12 h and CD38 protein at 6 h after LPS stimulation were significantly higher than those before (both P< 0.05). Conclusions:In both normal and endotoxin-tolerant THP-1 cells, LPS upregulates the expression of CD38, which is an NAD + digestive enzyme, and an indirect indicator of NAD + level in monocyte reinfection at the stage of immunosuppression. This study provides a preliminary reference for further investigation on the applicability of CD38 as a potential biological marker in the clinical diagnosis of neonatal sepsis.

Objective To investigate the role of KiSS-1 and matrix metalloproteinase(MMP)9 in trophoblasts in the pathogenesis of preeclampsia and their relation to perinatal outcome of neonates. Methods RT-PCR and western blot analyses were used to detect the MMP-9 and KiSS-1 expression levels in trophoblast of 40 patients with preeclampsia (15 cases of mild and 25 cases of severe preeclampsia)(preeclampsia group) and 20 cases of term pregnancy (normal pregnancy group) and their correlations with symptoms and perinatal outcome of neonates were analyzed. Results (1) The KiSS-1mRNA and metastin expression levels in trophoblasts of preeclampsia group were 1.73?0.24 (A value) and (78.4?8.0) ?g/ 100 ?g total protein separately,those of mild preeclampsia were (1.50?0.15) and (72.4?6.9) ?g/ 100 ?g total protein , and severe preeclampsia were (1.87?0.20) and (83.52?3.57) ?g/100 ?g total protein , which were all significantly higher than those of normal pregnancy group [1.24?0.25, P

Objective To establish a loop-mediated isothermal amplification(LAMP) quantitative method for rapid detection of Japanese encephalitis and dengue fever virus.Methods According to the LAMP principle,design primers for LAMP detection and reaction system,establish LAMP detection method,and to evaluate the linear relationship between initial copy number and the specificity,sensitivity,repeatability and the reaction time(fluorescence signal value of 1×104 corresponding time).Results One sets of LAMP primers could be used to complete the detection work in 0.5 h.The sensitivity of LAMP detection technology was 10 times higher than that of the classical PCR technology,and no cross reaction with other viruses,and the coefficient of variation of the average test was less than 5%.There was a good linear relationship between cycle threshold and template concentration.Conclusion This method has high specificity,sensitivity,simple operation,which is easy to get the results,low equipment requirements and rapid,suitable for primary health institutions and the field inspection agencies for wide applications.

Objective To study the effects of quantitative 24-hour urinary protein on the thyroid hormone levels in patients with severe preeclamp-sia,and clarify the impact of severe urinary protein on hypothyroid in severe preeclampsia patients. Methods A total of 166 patients with severe pre-eclampsia were recruited for the study and divided into mild proteinuria group(2.0-4.9 g/d),midrange group(5-10 g/d)and severe group(>10 g/d)according to the quantitative 24-hour urinary protein. 268 healthy female individuals with normal blood pressure and uric routine in the same stage of pregnancy and of the same age were selected into control group. Serum thyrotropin(TSH),free triiodothyronine(FT3)and free thyroxine (FT4)levels were determined by solid-phase chemiluminescent enzyme immunoassay method(CMIA). The thyroid peroxidase antibody(TPOAb) and thyroglobulin antibody(TGAb)concentration were detected by electrochemiluminescent assay(ECLIA). Results TSH levels were signifi-cantly higher in patients comparing to the control group(P<0.01). In addition,severe group showed higher TSH levels than mild group(P<0.01). FT4 and FT3 levels were obviously decreased with the progression of the disease(P<0.01 and P<0.05). The positive rate of TPOAb in mild group was significantly higher than that in moderate group(OR=9.8,P<0.05). There was no significant difference of the TGAb positive rate among three patient groups(P>0.05). The incidence of subclinical hypothyroidism and clinical hypothyroidism in severe group was significantly higher than that in mild group and in control group(OR=2.5,P<0.05 and OR=9.0,P<0.05;OR=8.0,P<0.01 and OR=43.4,P<0.01). Conclusion Our re-sults indicated that 24-hour urine protein in severe preeclampsia patients has extensive effects on thyroid hormones levels. With the increasing of quantitative 24-hour urinary protein,the level of TSH increased and the FT4 decreased. Thyroid autoantibody positiveness has extensive effects on 24- hour urine protein. Incidence of hypothyroid increased with the increase of quantitative 24-hour urinary protein. 24-hour urinary protein quantitative was a risk factor for hypothyroidism in severe preeclampsia patients. More attention should be paid to the monitoring of 24-hour urinary protein in se-vere preeclampsia patients.

Objective To observe and discuss the effect of the traditional Chinese drug complex prescription ShensuⅡfrom Xintong Changluo therapeutic principle on renal interstitial fibrosis (RIF) and transforming growth factor-β1 (TGF-β1) expresssion in focal segmental glomerulosclerosis (FSGS) model rats. Methods Forty-eight healthy male SD rats were randomly divided into control group (n=12) and modeling group (n=36). Rats of modeling group were injected by doxorubicin hydrochoride for FSGS model. Rats of modeling group were sub-divided into model group, benazepril group and TCM group randomly. In 12 weeks, TCM group was given by intragastric administration of ShensuⅡ(3.5 g/100 g), benazepril group was given by intragastric administration of benazepril suspension (0.33 mg/100 g), control group and model group were given by intragastric administration of same volume of saline. HE/Masson staining was used to observe changes of tubulointerstitial pathomorphology. The degree of injury and fibrosis was measured. The expressions of fibronectin (FN) and TGF-β1 were detected by immunohistochemical SP method. Results The process of renal interstitial fibrosis was slower in FSGS rats of TCM group. Renal interstitial pathological index was 1.51 ± 0.80 in TCM group, which was lower than that of model group (2.18 ± 0.38) and benazepril group (1.79 ± 0.24). The index of renal interstitial fibrosis was 2.39 ± 0.13 in TCM group, which was lower than that of model group (3.11 ± 0.25) and benazepril group (2.80 ± 0.41). The relative expression of FN in renal interstitial was 14.19 ± 3.06 in TCM group, which was lower than that of model group (21.25 ± 3.31) and benazepril group (18.51±2.29). The relative expression of TGF-β1 in renal interstitial was 2.64±0.21 in TCM group, which was lower than that of model group (6.02 ± 0.12) and benazepril group (3.79 ± 0.46). All the differences were statistically significant (P<0.05). Conclusion Xintong Changluo complex prescription ShensuⅡcan reduce the process of renal interstitial fibrosis in FSGS model rats, which may be related with the inhibiting expression of TGF-β1.

Objective To observe the clnical effects of influence of tanshinone type ⅡA sulfonate on preventing hepatic artery thrombosis after transplantation.Methods A total of 60 patients after liver transplantation were randomly individed into the treatment group and control group, each 30 patients. The treatment group received tanshinone ⅡA sodium sulfonate treatment (60 mg qd, ivgtt continuous 10d) , while the control group used conventional heparinization. The blood coagulation index and the thrombelastograph variables were detected after 7 days and the hepatic artery resistance index (RI) was detected by using Doppler ultrasonography. The postoperative complications and mortality rates were analyzed.Results Although it had little improvement on the coagulation function after liver transplantation, tanshinone ⅡA sodium sulfonate had significant improvement on the time of thrombelastograph parameters reaction (6.35 ± 1.59 minvs. 5.21 ± 1.37 min,t=2.453) and maximum amplitude (58.07 ± 5.42 mmvs. 61.67 ± 5.63 mm,t=-2.532). It showed that RI have significantly statistical difference between the two groups after treatment (0.73 ± 0.11vs. 0.62 ± 0.10;t=-2.948,P<0.01). During the trial, the control group had 2 cases of postoperative complications, HAT and bleeding.Conclusions The Tanshinone ⅡA sodium after liver transplantation can improve the clotting mechanism, preventing HAT.

Objective To investigate the relationship between tumor necrosis factor-alpha (TNF-α) and development of chronic obstructive pulmonary disease (COPD).Methods COPD patients and controls were divided into three groups: COPD group (n=66), smoker control group (n=42) and health control group (n=23). COPD group was further divided into the serious group (n=23) and non-serious group (n=43). The concentration of TNF-α of all cases was detected by human Th1/Th2 cytokine kit.Results The concentration of TNF-α in the COPD group was significantly higher than that of the smoker and healthy groups ( P<0.01). Furthermore, compared to non-serious COPD group, the concentration of TNF-α was higher in the serious COPD group ( P<0.05).Conclusion The concentration of TNF-α might be related with the pathogenesis and development of COPD.