Objective:To investigate the effects of anti-CTLA4 antibody on Th1/Th2 shift in unexplained recurrent spontaneous abortion. Methods:The levels of IL-2,IFN-?and IL-4 were assessed by ELISA in the supernatant of cultured maternal peripheral blood mnnonuclear cells(PBMC) from 30 patients of Unexplained RSA. Cytokine synthesis was induced by activation with antigens from a chnriocarcinoma cell line of JEG-3,with anti-CTLA4 antibodv or IgG. Results:We detected significantly greater levels of the Th1 cytokines IL-2、IFN-?(P0.05). Conclusion:Anti-CTLA4 antibody can up-regulate Th1-type cytokines and down-regulate Th2-type cytokines, which indicates that in URSA anti-CTLA4 antibody could swith T cell responses to Thl,and is harmful to URSA.

Objective · To investigate the effects of a p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 on biological function changes of human extravillous trophoblast cells induced by hypoxia/re-oxygenation (H/R). Methods · In-vitro cultured early pregnancy villus explants and human extravillous trophoblast cell line HTR8/SVneo were assigned to 4 groups according to different interventions, i.e. control group, SB203580 group (p38 MAPK inhibition), H/R group (simulation of preeclampsia by the oxidative stress model), and SB203580+H/R group. The effects of SB203580 on biological functions of human extravillous trophoblast cells under oxidative stress in early pregnancy villus explants were observed. Protein expression and phosphorylation of p38 MAPK in HTR8/SVneo cells were measured with Western blotting. Cell migration and invasion were observed with Transwell migration and Matrigel invasion assay, respectively. Gelatin zymography was used to measure the activity of matrix metalloproteinase (MMP) -2/9 in supernatant. ELISA was used to detect the levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng). Results · SB203580 could promote the exogenous migration of human extravillous trophoblast cells in early pregnancy villus explants under oxidative stress. H/R could decrease the migration and invasion of HTR8/SVneo cells , and increase the phosphorylation level of p38 MAPK in HTR8/SVneo cells and the secretion of sFlt-1 and sEng. SB203580 could increase the activity of MMP2/9 in supernatant and cell migration and invasion, decrease the phosphorylation level of p38 MAPK in HTR8/SVneo cells under oxidative stress and the secretion of sFlt-1 and sEng. Conclusion · SB203580 can protect biological functions of human extravillous trophobalst cells under oxidative stress.

OBJECTIVE: To analyze the uncertainty of determination of western medicine composition illegally added into Chinese patent drug and find out the effect factors of uncertainty in order to provide scientific basis for evaluating test report. METHODS: Antler tablets were tool as example. Qualitative analysis of additive composition was carried out using HPLC-MS method and the content of sildenafil citrate in each tablet was determined. Uncertainty of test was evaluated according to the regulation specified in Evaluation and Expression of Measurement of Uncertainty (JJF1059-1999). RESULTS: The uncertainty of the test is 0.10 mg in each tablet. CONCLUSION: The uncertainty of the experiment is mainly caused by non homogeneity of sample.

Objective · To investigate the effects of growth arrest and DNA damage 45 alpha (Gadd45α) on the migration and invasion function of human extravillous trophoblast cells under hypoxia/re-oxygenation (H/R). Methods · Human extravillous trophoblast cells were infected by shRNA lentivirus targeting Gadd45α gene, to knock down Gadd45α gene expression. Then the oxidative stress model of preeclampsia was used in vitro to observe the changes of cell biological functions. The experiments were divided into 4 groups, nontreated group, hypoxia/re-oxygenation group, shRNA Gadd45α+H/R group and shRNA negative control+H/R group. Human villous explant experiments were used to determine the effects of silencing Gadd45α on human extravillous trophoblast cell under oxidative stress. Protein expression of Gadd45α was identified by Western blotting. Changes of cell migration and invasion were detected by transwell migration and Matrigel invasion assay. Gelatin zymography was used to detect the expression of matrix metalloproteinase (MMP) -2/9 in culture medium. Results · Hypoxia/re-oxygenation can increase the expression of Gadd45α in HTR8/SVneo cells and damage the trophoblast cell migration and invasion. Knocking down Gadd45α can increase the activities of MMP2/9, which can increase the cell migration and invasion. Conclusion · Knockdown of Gadd45α gene has promoted cell migration and invasion function of human extravillous trophobalst cells under oxidative stress.

Objective To investigate the value of apparent diffusion coefficient(ADC)of breast carcinoma before and one course after neoadjuvant chemotherapy,and to predict the response of breast carcinoma to neoadjuvant chemotherapy.Methods Twenty female patients with 21 breast carcinoma lesions were examined with diffusion weighted imaging(DWI)and contrast-enhanced MRI within 7 days prior to neoadjuvant chemotherapy(adriamycin + taxinol),during 18-21 days after the first couse of therapy and within 7 days prior to the surgery resection.The tumors were divided into response group and relative nonresponse group by change of the tumor.The difference of ADC between the two groups before and after 1st course of chemotherapy was compared.Results In chemotherapy response group with 15 lesions,the mean ADC was(0.98±0.15)×10-3 mm2/s before chemotherapy and increased to(1.22±0.23)×10-3 mm2/s after 1 st course of chemotherapy(P＜0.05),while in nonresponse group with 6 lesions,the mean ADC before and after chemocherapy was(1.09±0.08)×10-3 mm2/s and(1.11±0.07)×10-3 mm2/s,respectively,with no statistical difference(P>0.05).The mean ADC of response group prior to chemotherapy was significantly lower than that of nonresponse group(P＜0.05).A significant negative correlation(r=-0.51,P＜0.05)was observed between the ADC prior to treatment and change of ADC after the first couse therapy.Conclusion Preliminary results revealed the initial ADC of the tumor in patient with breast carcinoma and the changes of ADC after 1 st couse treatment may predict response to neoadjuvant chemotherapy.

PurposeTo investigate a multi-parametric protocol for breast MRI examination and lesions assessment correlated to the American College of Radiology (ACR) breast imaging reporting and data system (BI-RADS) categorization, and to improve the management of the breast lesions.Materials and Methods 301 pathologically confirmed lesions on 278 patients were retrospectively included. The scan protocol used a dynamic contrast enhancement sequence (DCE) of 1 mm×1 mm×1 mm spatial resolution, 120 temporal resolution and a diffusion weighted imaging (DWI) of b=1000 s/mm2. The malignant morphological features on the early-enhanced images, type II or III time intensity curve and the apparent diffusion coefficient (ADC) value less than benign/malignant threshold was equally weighted. Each was given 1 point when present malignant features and treated different on mass and non-mass-like enhancement lesions. When the sum of score was ≥2 points, the lesion was categorized as BI-RADS 5. When the sum of score was 1 point, the lesion was categorized as BI-RADS 4. When the sum of score was <1 point, the lesion was categorized as BI-RADS 3. The other specific benign findings were categorized as BI-RADS 2. No abnormality on DWI, DCE, T2WI and T1WI was categorized as BI-RADS 1. The final categories were correlated to the pathological grades as benign (B), high risk (HR) and malignant (M).Results When grouped HR as malignant (M+HR), the area under curve (AUC) of the ROC was 0.860. When grouped HR as benign (B+HR), the AUC of the ROC was 0.876, and the optimized sensitivity, specificity and accuracy was 85.3%, 86.8% and 85.1%, respectively, which were better than the other grouping. If the management of HR lesions could be lumptoectomy or short-term follow-up, the positive predictive value (PPV) of BI-RADS 5 for excisable lesions (M+HR) was 93.2%, the PPV of BI-RADS 4 for excisable lesions (M+HR) was 46.9% and the biopsy was essential. The PPV of BI-RADS 3 and below for follow-up lesions (B+HR) was 90.4%.Conclusion A simple diagnosis algorithm was established, which equally weighted the DCE morphological feature, DCE-TIC and DWI-ADC. The diagnosis protocol was well consistent with BI-RADS categorization and could predict the benign, high risk and malignant lesions in pathology as well as the proper management.

Objective To explore the role of specific AT rich sequence binding protein 1(SATB1) and wnt/β-catenin signaling pathways in the regulation of trophoblast invasion and its effect in the pathogenesis of preeclampsia. Methods From March 2013 to March 2014, 20 cases of human villous tissues (early pregnancy group) from women of 8-10 gestational weeks who received artificial abortion at the First Affiliated Hospital of Chongqing Medical University, 18 cases of placental tissues (mid-pregnancy group) from women of 18-20 gestational weeks who had labor induction by water bag, 20 cases of placental tissues (normal full-term group) from healthy full-term pregnancy women and 20 cases of placental tissues (preeclamptic group) from women with preeclampsia who received elective c-section in were collected. Immunohistochemical SP method was utilized to determine the position of SATB1 and beta-catenin in villous tissues or placental tissues. Western blot was performed to analyze the expression level of SATB1 and beta-catenin in villous tissues or placental tissues. Immunofluorescence assay was used to determine the location of SATB1 andβ-catenin in HTR8/SVneo cells. Western blot was performed to detect the expression level of SATB1 and beta-catenin in HTR8/SVneo cells cultured in normoxia and hypoxia reoxygenation(H/R) condition. Co-Immunoprecipitation detection was used to evaluate the interaction between SATB1 andβ-catenin in placental tissues in preeclamptic group and HTR8/SVneo cells in H/R group. Gelatin zymography analysis was used to measure the activity of matrix metalloproteinases(MMP)-2 and 9 in placental tissues from preeclamptic group and HTR8/SVneo cells in H/R group. Results (1) In the normal full-term group, rare syncytiotrophoblastic nodule, less fibrinoid necrosis and abundant numbers of capillary could be observed in placental tissues. In comparison, there were obvious vacuolation in the cytoblast of syncytiotrophoblast, rich fibrinoid necrosis and poor numbers of villous capillary in placental tissues from preeclamptic group. (2) SATB1 could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of SATB1 were more weakening in preeclamptic group than in the normal full-term group. (3) β-catenin could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of β-catenin were more weakening in preeclamptic group than in the normal full-term group. (4) The protein expression levels of SATB1 in early pregnancy group, mid-pregnancy group, normal full-term group and preeclamptic group were 0.300 ± 0.009, 0.271 ± 0.015, 0.238 ± 0.018 and 0.153 ± 0.007, respectively. The protein levels of β-catenin among the above groups were 0.743±0.041, 0.648±0.021, 0.549±0.069 and 0.269±0.047, respectively. Both the expression of SATB1 andβ-catenin protein were significant decreased in placental tissues from preeclamptic group compared with the other three groups. (5) The SATB1 andβ-catenin protein was located in nucleus of trophoblast and a small amount was in the cytoplasm. The fluorescence intensity of both SATB1 and β-catenin in the H/R group were significantly decreasing when compared to the normoxia group. (6) HTR8/SVneo cells in H/R group showed a significant decrease in both SATB1 andβ-catenin protein levels when compared to the normoxia group. The protein level of SATB1 in the normoxia group was 0.213 ± 0.005, while was 0.083 ± 0.021 in the H/R group. The protein level ofβ-catenin in the normoxia group was 0.797±0.081, and was 0.543±0.131 in the H/R group. (7) There was an interaction between SATB1 and β-catenin in placental tissues from the preeclamptic group and HTR8/SVneo cells exposed by H/R. (8) The enzymatic activity of MMP-2 and MMP-9 protein were decreased significantly in placental tissues from the preeclamptic group (2.251±0.310, 1.447 ± 0.102, respectively) when compared to the normal full-term group (7.098 ± 0.451, 5.502 ± 0.197, respectively). MMP-2 and MMP-9 were significantly decreased in the H/R group (0.471 ± 0.104, 0.297 ± 0.103, respectively) when compared to the normoxia group (0.842 ± 0.209, 0.595 ± 0.100, respectively). Conclusion The expression of SATB1 decreased in the placenta of preeclampsia. This may influence the activity of MMP-2 and 9 by regulating Wnt/β-catenin signaling pathways, affect trophoblast invasion and eventually result in preeclampsia.

OBJECTIVE:To establish a method for the determination of related substances in bisacodyl raw material and enteric-coated tablet. METHODS:HPLC was performed on the column of Hibar C18 with mobile phase of acetonitrile-20 mmol/L ammonium acetate (acetic acid adjust pH to 5.0)(55∶45,V/V),detection wavelength was 265 nm,flow rate was 1.0 ml/min, column temperature was 30℃,and the injection volume was 20 μl. RESULTS:The linear range of bisacodyl was 0.25-5.0 mg/ml (r=0.999 9);the limits of detection and quantification were 19-25 ng and 61-68 ng for bisacodyl and impurity A,B,C,D and E;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 99.50%-101.00%(RSD=0.5%,n=9). CONCLUSIONS:The method is specific, sensitive and reproducible, and can be used for the determination of related substance in bisacodyl raw material and enteric-coated tablet.

Objective To explore the optimal acupuncture scheme in intervening hypermyotonia of the affected limbs in stroke.Method Eighty eligible stroke patients were randomly grouped by using the orthogonal experimental design, by adopting electroacupuncture (A), daily acupuncture frequency (B), and acupoint (C) as three factors, and two levels. A total of 20 treatment sessions were conducted. Each group was scored by using the Modified Ashworth Scale (MAS) before and after the treatment, and the safety was also evaluated.Result There was a significant difference in comparing the MAS score between the two different levels of factor A (P<0.05), while there were no significant differences in comparing the MAS scores between different levels of factor B and factor C (P<0.05). Concerning the improvement of the MAS scores of wrist, elbow, knee, and ankle joints, A2B1C1(i.e. electroacupuncture twice a day with acupoints from yin meridians) was the optimal treatment scheme.Conclusion Electroacupuncture twice a day is the optimal treatment scheme for hypermyotonia of the affected limbs in stroke, as it can effectively ease the hypermyotonia, improve the function and symptoms of the limbs, and has a satisfactory security evaluation.

Objective To discuss the adjustment effects of kidney reinforcing medicine on growth and development and the content of choline acetyl transferase (CHAc) in rats with kidney-deficiency constitution. Methods The method of cats scare rats was used to build composite offspring rat models with deficiency and acquired dystrophy, and then the models were divided into model group, Zuogui Pill group and Yougui Pill group. The rats in blank group came from normal pregnant rats. Baby rats were scared and received gavage at the same time. All administration groups received suspension of Zuigui Pills or Yougui Pills. Blank group and model group received the same amount of normal saline, once a day, for 3 months. Growth and development of rats were observed. Weight and food utilization of rats aged 5–8 weeks in each group were recorded. The content of brain CHAc was detected by ELISA. Results When offspring were 5–8 weeks old, weights of rats in the model group were lower than blank group (P<0.01); while weights of rats in Zuogui Pill group and Yougui Pill group increased significantly (P<0.05) and the food utilization was positively correlated with weight; the CHAc content in the model group decreased significantly compared with blank group (P<0.05, P<0.01); while the CHAc content in Zuogui Pill group and Yougui Pill group increased significantly compared with the model group (P<0.01). Conclusion Kidney reinforcing medicine can improve backward growth and development of rats with kidney-deficiency constitution and adjust CHAc in the brain, so as to promote the learning and memory ability of rats with kidney-deficiency constitution.