Objective To investigate the effect of extracellular signal-regulated kinase(ERK)pathway OH nitriC oxide(NO)release by human umbilical vein endothelial cell(HUVEC)induced by placental growth factor-1(PLGF-1).Methods During January to April 2006,50 samples of umbilical vein blood were collected from newborns delivered by cesarean section due to intrauterine distress and abnormal fetal position.HUVEC were primarily cultured by trypsin digestion.Then the following procedures were performed:(1)Cells were identified using the morphology andⅧfactor immunohistochemistry methods if the culture WaS satisfactory.(2)Cells were collected,and fms.1ike tyrosin kinase(Fit-1)protein and its mRNA expression were detected with immunoprints and RT-PCR methods.(3)The protein wag extractedafter cells were treated with PLGF-1(cells were collected before the treatment and 2.5,5.10,20 min after the treatment).The protein levels of ERK were determined by immunoprints.(4)The cells were cultured witll serum-free culture medim containing PLGF-1 only(culture media were collected 20,40,160,360.480.720 and 1440 rain after the treatment).The quantity of NO was detected with nitrate reductase metllod(5)The ceHs were cultured with serum.free culture medium containing PI)98059,the inhibitor of mitogen-activated protein kinase(MAPK)/MEK for 60 min Then the cells were cultured continuously with the serum-free culture medium containing PLGF-1 for 60 mira The culture media were coilected.The quantity of NO was detected by nitrate reductase method.The samples were divided into treatment group and control group.Control group was exactly the satne in treatment time,culture condition,and time to colleet the cells as the treatment group.except that it WaS not treated with PLGF-l or PD 98059.Resuits (1)By morphology and Ⅷ factor inununohistochemistry the cultured cells were identified to be HUVEC.(2)Fit-1mRNA and protein were expressed in HUVEC.(3)Expression of ERK protein started to increaSe at 2.5 min after treatment of HUVEC with PLGF-1,reaching the peak at 5 min,and decreased at 10 min.(4)Incomparison with the control group.NO started to increase at 20 rain after treatment of HUVEC with PLGF-lat 480 min(15.82±0.69)μmol/L Comparison between the two groups showed a significant difference (P<0.05).(5)ReleaSe of NO from the cells treated with PD98059 for 1 hour and PLGF WaS significantly inhibited,compared with the ceils treated with PLGF-1 only.Conclusion ERK pathways play an important role in N0 release bv HUVEC induced by PLGF.

The expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) in the placenta of the patients with pregnancy induced hypertension (PIH) was detected and its role in the pathogenesis of PIH was studied. The pathological changes in placental vessels were observed by HE staining. NO2-/NO3-, the stable metabolic end products of NO, was measured with nitrate reductase. The eNOS activity in placental tissues was assayed by spectrophotometry. Western blot analysis was applied to detect NOSTRIN expression. The incidence of thickening and fibronoid necrosis of placental vessels was significantly higher in women with PIH than in the normal group (P < 0.01). The levels of placental NO2-/NO3- in PIH patients (27.53 +/- 7.48 micromol/mg) were significantly lower than in normal group (54.27 +/- 9.53 micromol/mg, P < 0.01). The activity of eNOS was significantly decreased in PIH group (12.826 +/- 3.61 U/mg) as compared with that in normal group (21.72 +/- 3.83 U/mg, P < 0.01). Western blot analysis revealed that both groups expressed 58 kD NOSTRIN, but the protein level was significantly higher in women with PIH than in the normal group (P < 0.01). A significant negative correlation existed between the expression of NOSTRIN protein and the activity of eNOS in placental tissue of women with PIH (r = -0.57, P < 0.01). It was concluded that the level of NOSTRIN expression in placenta of women with PIH was increased, which may play an important role in the pathogenesis of PIH.

Objective:This study aimed to detect the expression of annexin A1 in human non-small cell lung cancer (NSCLC) and to explore its clinical significance. Methods:We detected the expression levels of annexin A1 in NSCLC and the same patient's dis-tal cancerous tissue (from the edge of the tumor >5 cm) by real-time fluorescence quantitative polymerase chain reaction (real-time PCR), Western blot, and immunohistochemical methods, and then analyzed their correlation with clinical pathological parameters. Re-sults: Real-time PCR showed that the expression of annexin A1 mRNA in NSCLC was higher than that of distal cancerous tissue (0.574±1.403 vs. 0.240±0.893, t=2.060, P=0.045). Moreover, the expression of annexin A1 in NSCLC was associated with the degree of differentiation, lymph node metastasis, and TNM staging (P<0.05), but independent of gender, age, smoking history, tumor size, and histological type (P>0.05). Western blot and immunohistochemistry revealed that the expression of annexin A1 protein in NSCLC was higher than that of distal cancerous tissue. Conclusion:Annexin A1 is highly expressed in the cancer tissues of patients with NSCLC, which may be correlated with the occurrence and development of tumor and its invasion and metastasis.

By combining frequency modulation Chirp Z transform ion mobility spectrometers (IMS) and multi nozzle electrospray array ionization source, a method of NanoESI-Chirp Z transform ion mobility spectrometry-high performance liquid chromatography was developed for the determination of n-alkyl ammonium bromide compounds.The parameters of NanoESI-Chirp Z transform IMS such as electric field intensity, solvent composition, and solution flow rate were investigated and optimized.Subsequently, four kinds of n-alkyl ammonium bromide compounds were respectively detected by this developed Chirp Z transform method and Fourier transform method, and the obtained results were compared.The result indicated that the optimum conditions were electric field intensity of 4.5 kV, and ESI solution flow rate of 8 μL/min.Then a test mixture containing tetrabutylammonium bromide, tetrapentylammoniumbromide, tetrahexylammonium bromide, tetraheptylammonium bromide, tetranoctylammoniumbromideandtetrakis(decyl) ammonium bromide was successfully separated and determined by the HPLC-nanoESI-Chirp Z IMS method.Chirp Z transform method provided higher signal to noise ratio compared to conventional signal averaging method, and was superior to FT method in the determination of drift time.

To investigate the expressions of placental growth factor (PLGF) in placenta with hypertensive disorders of pregnancy (HDP), 45 women with HDP and 20 normally pregnant women were studied. Among 45 women with HDP, there were 23 cases of severe preeclampsia and one case of eclampsia. The location and level of PLGF proteins was determined by immunohistochemistry and Western blot. The expression of PLGF mRNA in placenta was assessed by reverse transcriptional-polymerase chain reaction (RT-PCR). The results showed that: (1) The distribution of PLGF in placenta with HDP was similar to normal one, which was mainly in the cytoplasm of villous syncytiotrophoblast and villous stroma; (2) The expression of PLGF protein was significantly decreased in placentas with mild and severe preeclampsia compared to the normal ones (0.3 +/- 0.4 vs 0.6 +/- 0.4, 0.2 +/- 0.5 vs 0.6 +/- 0.4, P < 0.01). There were no differences between the gestational hypertension placenta and normal one (0.5 +/- 0.6 vs 0.6 +/- 0.4, P > 0.05); (3) The transcription levels of the PLGF mRNA in placentas with preeclampsia were significantly lower than in normal groups (3.33 +/- 0.39 vs 4.87 +/- 0.60, 1.97 +/- 0.29 vs 4.87 +/- 0.60, P < 0.01), and no differences were found between the gestational hypertension placenta and normal groups. These findings suggest that the abnormal expression of PLGF in placentas is related to the pathogenesis of HDP.
Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy/*metabolism ; Pregnancy Proteins/*biosynthesis ; Pregnancy Proteins/genetics

In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P < 0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P < 0.05). No significant difference in the Pin1 expression was found between disease stages (FIGO), pathological grades or pelvic lymph node metastasis status (P > 0.05). The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix (P < 0.05). In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P < 0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.
Cervical Intraepithelial Neoplasia/metabolism ; Ki-67 Antigen/*biosynthesis ; Ki-67 Antigen/genetics ; Peptidylprolyl Isomerase/*biosynthesis ; Peptidylprolyl Isomerase/genetics ; Tumor Markers, Biological ; Uterine Cervical Neoplasms/*metabolism

Objective To investigate the feasibility of applying nanotechnology against M.tuberculosis growth.Methods Chitosan-antisense ODN nanoparticles were prepared by complex coacervation method.Varied amount of ODNs and chitosanODN nanoparticles were added to the cultures and the growth of the bacilli was monitored.Results The nanoparticles were composed of(35.6±0.9)% ODN and(64.4±0.9)% chitosan.Compared to the free ODN,antisense nanopartilces were more effective in inhibiting the proliferation of M.tuberculosis.Antisense nanoparticles decreased growth by(2.8±0.1)CFU/ml at concentration of 4/μmol/L.Conclusion Chitosan-ODN nanoparticles were more effective in inhibiting the growth of M.tuberculosis than free ODN.

Objective To observe the effects of herb-partition moxibustion at navel on the expressions of Bcl-2 and Bax in rats with premature ovarian failure (POF); To explore its mechanism underlying improvement of POF. Methods POF model was established by intraperitoneal injection of cyclophosphamide. Forty-eight Female Wistar rats were randomly divided into normal group, model group, moxibustion group and Western medicine group, with 12 rats in each group. Rats of moxibustion group were treated with herb-partition moxibustion at navel (Shenque, CV8), 30 min/d, once every other day; rats of Western medicine group were given intragastric dose of estradiol valerate, once a day, for 14 d. The protein and mRNA expressions of Bax and Bcl-2 in ovarian granulosa cells were detected by immunohistochemal method, Western blot and qRT-PCR, respectively.Results The weights of ovaries in the model group rats were significantly lower than those in the normal group (P<0.05). The expressions of Bcl-2 and Bax protein in ovarian tissue of each group were mainly expressed brown coloration and in the cytoplasm of cells. The range of Bax protein in granulosa cells of moxibustion group and Western medicine group was significantly less than that in model group. The range of Bcl-2 in moxibustion group and Western medicine group was significantly larger than that in model group. Compared with the normal group, the expressions of Bcl-2 protein and mRNA in the model group rats significantly decreased (P<0.05,P<0.01), and the expression of Bax protein and mRNA significantly increased (P<0.05,P<0.01). The expression level of Bcl-2 protein and mRNA was significantly higher in the moxibustion group and in the western medicine group than that in the model group (P<0.05,P<0.01). The expression level of Bax protein and mRNA in the moxibustion group was significantly lower than that in the model group (P<0.05,P<0.01). ConclusionThe herb-partition moxibustion at navel can improve the function of ovaries by up regulating the expression of Bcl-2 and down regulating the expression of Bax in POF rats model.

Objective To survey the incidence of sleep disorders among patients with industrial injuries and analyze the relevant factors.MethodsA total of 112 depressed patients ( male 106,female 6; aged 22-79 years,course of disease 35 d-25 years) were assessed by a professional psychologist using life satisfaction index A,the type A behavior pattern scale,the Pittsburgh sleep quality index (PSQI) and the Barthel index.ResultsThe incidence of disordered sleep was 40.18％ (45/112),of whom mildly depressed patients were 34.29％,moderately depressed patients 42.86％ and severely depressed patients 85.71％.The incidence of sleep disorders increased with increasing depression severity.The incidence of disordered sleep was significantly higher among the severely depressed patients than among those mildly or moderately depressed,but there was no significant difference in incidence between moderately and mildly depressed patients.PSQI scores among the severely and moderately depressed patients were significantly higher than among those mildly depressed,but there was no statistically significant difference in average PSQI scores between the severely and moderately depressed patients.The sleep disorder group suffered significantly poorer sleep quality and took significantly longer to get to sleep.There were no significant differences in average age,educational level,marital status,social relations,family and social support,gender distribution or course of disease between the two groups.There were,however,significant differences in family income,life satisfaction,character type and disease species between the groups.ConclusionThe incidence of disordered sleep among depressed patients after industrial injury is correlated with the severity of depression,family income,life satisfaction,the type of injury and the patient's character.