[Comparison of the methods for detecting NTRK gene fusion variations in papillary thyroid carcinoma].

Objective: To investigate the frequency of neurotrophic tyrosine receptor kinase (NTRK) gene variations in papillary thyroid carcinoma (PTC) and to analyze the feasibility of detecting tropomyosin receptor kinase (TRK) proteins using immunohistochemistry (IHC) to predict the fusion variation of NTRK. Methods: A cohort of 848 PTC cases was collected at the Department of Pathology, Shenzhen People's Hospital from June 2017 to June 2020. The expression levels of TRK proteins were detected using IHC in 848 PTC samples, and the DNA-based next generation sequencing (NGS) was performed to detect NTRK rearrangements in 150 PTCs. Results: There were 242 males and 606 females, with an age range of 9-83 years. In 120 cases with TRK expression detected by IHC, 13 cases were confirmed to harbor a NTRK gene fusion by NGS. The frequency of NTRK fusion in PTC was 1.5% (13/848). The sensitivity and specificity of TRK-IHC positivity for screening NTRK fusion in PTC were 100% and 21.9%, respectively. The specificity of weak-, moderate- and strong-positive stains of TRK IHC were 23.8%, 76.9% and 93.8%, respectively. The specificity of NTRK gene fusion was predicted to increase with the enhanced intensity of IHC staining. In BRAF V600E negative PTC samples, the specificity of weak-and moderate-positive stains of TRK IHC increased to 62.5% and 96.8%, respectively. Seven NTRK fusion partners were found in the PTC, including EML4, ETV6, CDH1, GJD2, TPR, TFG and SQSTM1. Conclusions: There is a low variation frequency of NTRK gene fusion in PTC. TRK IHC can be used as a screening method for NTRK fusion variation in PTC. The specificity of TRK IHC predicting NTRK fusion can be further enhanced by increasing the cutoff value of the positive cell number and staining intensity of TRK-IHC staining, or being combined with BRAF V600E negativity.

[Prevalence of intestinal protozoan infections among rural children in Henan Province from 2014 to 2015].

OBJECTIVE: To investigate the prevalence and influencing factors of intestinal protozoan infections among rural children in Henan Province. METHODS: A total of 104 survey sites were sampled from 35 counties (cities) in Henan Province using the stratified cluster sampling method to investigate the prevalence of intestinal protozoan infections among rural children from 2014 to 2015. The trophozoites and cysts of intestinal protozoa were identified using the iodine staining method and the physiological saline direct smear method (one detection for one stool sample). The prevalence of intestinal protozoan infections was compared among rural children with different characteristics, and the factors affecting intestinal protozoan infections among rural children were identified. RESULTS: The overall prevalence of intestinal protozoan infections was 0.60% (40/6 771) among rural children in Henan Province from 2014 to 2015. There were 7 species of intestinal protozoa identified, and there was no species-specific prevalence (χ2 = 37.732, P = 0.000). No significant differences were found in prevalence of intestinal protozoan infections among rural children in terms of gender (χ2 = 1.793, P = 0.181), age (χ2 = 1.443, P = 0.486), occupation (χ2 = 0.219, P = 0.896) or ecological region (χ2 = 1.700, P = 0.637). In addition, terrain (χ2 = 2.311, P = 0.510), economic level (χ2 = 4.322, P = 0.229), source of drinking water (χ2 = 0.731, P = 0.393), eating raw vegetables (χ2 = 1.134, P = 0.287) and deworming (χ2 = 1.089, P = 0.297) had no remarkable effects on the prevalence of intestinal protozoan infections among rural children in Henan Province; however, the prevalence of intestinal protozoan infections varied significantly among rural children living in regions with different coverage of non-harmless toilets (χ2 = 10.050, P = 0.018). CONCLUSIONS: The prevalence of intestinal protozoan infections is low among rural children in Henan Province.

[Prevalence and influencing factors of intestinal parasitic diseases among rural children in Henan Province].

OBJECTIVE: To understand the epidemic status and influencing factors of intestinal parasitic diseases among rural children in Henan Province. METHODS: According to the Scheme for The National Survey on Current Status of Major Human Parasitic Diseases in China, the survey counties were selected based on the ecological zones and economic levels in Henan Province between 2014 and 2015. Then, the included counties were stratified according to the topography and economic levels. A township was randomly sampled from each stratum, and a village was randomly sampled from each township as the study site. Finally, a total of 104 study sites from 35 counties were enrolled for the survey of intestinal parasitic diseases in children. At least 250 fresh stool samples were collected from each study site for detection of intestinal helminth eggs with the Kato-Katz technique, for the identification of Necator americanus and Ancylostoma duodenale with the fecal culture method, and for the detection of intestinal protozoa trophozoite and cyst with the physiological saline smear and iodine staining techniques. In addition, the Enterobius vermicularis and tapeworm eggs were detected in children aged 3 to 6 years using the adhesive cellophane-tape perianal swab method. RESULTS: The overall prevalence of intestinal parasitic infections was 3.21% (214/6 671) among rural children in Henan Province, and the prevalence of intestinal helminthes (2.62%, 175/6 671) was higher than that of intestinal protozoa (0.60%, 40/6 671). A total of 12 species of intestinal parasites were found, including 4 nematodes species, one trematode species, and 7 protozoa species, and the highest infection was seen in E. vermicularis (2.47%, 161/6 671). Among the four ecological zones in Henan Province, the greatest prevalence of intestinal parasitic infections was detected among children in the Qinba Mountain Ecological Zone (5.85%, 90/1 538). There was no gender-specific difference in the prevalence of intestinal parasitic infections in children (P > 0.05); however, there were age- (χ2 = 32.762, P < 0.05) and education level-specific differences in the prevalence of intestinal parasitic infections in children (χ2 = 67.507, P < 0.05), with the greatest prevalence of E. vermicularis infection seen in all species of intestinal parasites in children at all age groups. Multivariate non-conditional logistic regression analysis showed that high education level, high coverage of harmless toilets, drinking tap water and deworming were protective factors for intestinal parasitic infections in children in Henan Province. The overall prevalence of intestinal parasitic infections appeared a tendency towards a gradual decline among children in Henan Province as compared to the previous two surveys. CONCLUSIONS: The overall prevalence of intestinal parasitic infections shows a tendency towards a remarkable decline among children in Henan Province. E. vermicularis infection should be given a priority for future parasitic disease control activities among rural children in Henan Province.

Preliminary results indicate increased expression of miR-184 in patients with renal carcinoma.

OBJECTIVE: Renal carcinoma is the second most common cancer in the urinary system with an increasing trend. The major treatment for renal carcinoma is surgery, which results in unfavorable prognosis at times. As a tissue-specific marker for tumor, microRNA (miR) exerts its functions via facilitating oncogenic gene expression or suppressing tumor suppressor gene. MiR-184 is known to be abnormally expressed in various tumors. There are few studies about the lack of miR-184 expression in renal carcinoma. PATIENTS AND METHODS: Real time-Polymerase Chain Reaction (PCR) was used to measure the expression of miR-184 in 38 renal carcinoma and adjacent tissues. The in vitro cultured renal carcinoma cell line ACHN was transfected with miR-184 mimic or inhibitor. The expression of miR-184 was measured by real time-PCR, and the cell proliferation was measured by MTT assay. The cell colony formation was examined, and the cell invasion potency was assessed by transwell assay. The apoptotic activity was measured by flow cytometry, and the Western blot detected protein expression change of ß-catenin/TCF3 pathway. RESULTS: Compared to tumor-adjacent tissues, miR-184 and ß-catenin/TCF3 showed an elevated expression in renal carcinoma tissues which were further increased with elevated RC stages (p<0.05). The transfection of miR-184 mimic into ACHN cells increased its expression, enhanced ACHN cell proliferation, colony formation, inhibited apoptosis, promoted tumor cell invasion, and increased the expression of ß-catenin and TCF4 proteins (p<0.05 compared to NC control group). CONCLUSIONS: MiR-184 is up-regulated in renal carcinoma tissues. The downregulation of miR-184 in renal carcinoma cells could facilitate cell apoptosis and inhibited tumor proliferation or invasion possibly via modulating ß-catenin/TCF4 pathway.

[Correlation between single nucleotide polymorphisms of rs4778137 located in OCA2 gene and clinical response of breast cancer patients receiving neoadjuvant chemotherapy].

Objective: To evaluate the correlation between single nucleotide polymorphisms (SNPs) of rs4778137 located in OCA2 gene and clinical response of breast cancer patients receiving neoadjuvant chemotherapy. Methods: A total of 140 breast cancer patients receiving neoadjuvant chemotherapy were enrolled to detect DNA in blood sample by DNA extraction kit and the rs4778137 polymorphism by sequenom. The relationship between SNPs of rs4778137 and pathologic complete response (pCR) were analyzed. Results: The frequency of CC, GC and GG genetype of rs4778137 was 48.6%, 31.4% and 20.0%,respectively. Thirty cases (21.4%) achieved pCR with CC allele in 9 cases(13.2%),GC allele in 10 cases (22.7%) and GG allele in 11 cases (39.3%),respectively,with a statistically significant difference(P<0.05). When conducting stratified analysis in accordance with the estrogen receptor (ER) status,only in ER negative group pCR was significantly associated with SNPs of rs4778137 (P<0.05). SNPs of Rs4778137, targeted therapy,subtypes,tumor stage were independent predictors of pCR in multivariate logistic regression analysis (P<0.05),and SNPs of rs4778137 was an independent predictors of pCR in ER negative group (P<0.05), but not in ER positive group group (P>0.05). Conclusion: SNPs of rs4778137 was associated with pCR only in ER negative patients receiving neoadjuvant therapy, and breast cancer patients with the GG allele were more likely to achieve pCR.

[Prognostic value of total cholesterol content of erythrocyte membranes in patients with acute coronary syndrome].

Objective: Previous cross-sectional studies suggested that elevated levels of total cholesterol content of erythrocyte membrane (CEM) could significantly increase the risk of acute coronary syndrome (ACS). The purpose of the present study was to assess the predictive value of baseline CEM levels for the risk of clinical endpoint events in patients with ACS through prospective follow-up studies. Methods: This study is a prospective follow-up study, which consisted of 859 patients with first ACS (698 patients with unstable angina pectoris and 161 patients with acute myocardial infarction), diagnosed and hospitalized in the First and Second Affiliated Hospital of Anhui Medical University. The routine blood lipid levels and CEM were measured. Patients were divided into two groups according to the median of baseline CEM: CEM≤131.56 µg/mg group (n=430) and CEM>131.56 µg/mg group (n=429). Patients were followed up at 6 months interval. The clinical endpoints were nonfatal myocardial infarction, nonfatal stroke, all-cause mortality, all-cause mortality, heart failure requiring hospitalization, and coronary artery revascularization. Kaplan-Meier curve analysis and Cox proportional hazard model were used to analyze the impact of elevated CEM on the occurrence of clinical end-point events. HR values and 95%CI of each variable were obtained. Cox regression analysis of all-cause mortality was performed according to whether patients had risk factors for coronary heart disease (hypertension, diabetes, smoking and elevated LDL-C) and whether they were treated with PCI. Results: The follow-up time was 1 640 (1 380, 2 189) days. Cox analysis after adjustment showed that an elevated baseline of CEM (>131.56 µg/mg) was associated with an increased risk of all-cause mortality (HR=1.690, 95%CI 1.041-2.742, P=0.034), but had no significant predictive effect on the other clinical endpoints. Subgroup analysis showed that elevated baseline CEM levels in ACS patients with LDL-C>1.8 mmol/L (HR=1.687, 95%CI 1.026-2.774, P=0.039), receiving in-hospital PCI (HR=2.365, 95%CI 1.054-5.307, P=0.037), or male (HR=1.794, 95%CI 1.010-3.186, P=0.046) were associated with an increased risk of all-cause mortality. Conclusion: The results showed that elevated CEM levels can increase the risk of all-cause mortality in ACS patients.

[Stratified sampling survey of major human parasitic diseases in Henan province].

Objective: To understand the prevalence of major human parasitic diseases and related factors in Henan province. Methods: This stratified sampling survey was carried out according to the requirement of national survey protocol of major human parasitic diseases, 2014-2015. The prevalence of soil-transmitted helminths infection, taeniasis and intestinal protozoiasis were surveyed in 104 sites selected from 35 counties (districts) and the prevalence of clonorchiasis was surveyed in 62 sites selected from 37 townships. In each survey spot, 250 persons were surveyed. A total of 26 866 persons and 15 893 persons were surveyed. Modified Kato-Katz thick smear was used to detect the eggs of intestinal helminthes. Tube fecal culture was used to identify the species of hookworm. The Enterobius eggs were detected in children aged 3 to 6 years by using adhesive tape. The cyst and trophozoite of intestinal protozoa were examined with physiological saline direct smear method and iodine stain method. Results: The overall infestation rate of intestinal parasites was2.02% in Henan, and the worm infection rate was higher than protozoa infection rate. Fourteen kinds of intestinal parasites were found, including nematode (5 species), trematode (2 species), and protozoan (7 species). The infection rate of Enterobius vermicularis was highest, and Qinba Mountain ecological area had the highest infestation rate of intestinal parasites in 4 ecological areas of Henan. There was no significant difference in intestinal parasite infection rate between males and females (χ(2)=3.630, P=0.057), and the differences in intestinal parasite infection rate among different age groups had significance (χ(2)=124.783, P=0.000 1). The infection rate reached the peak in age group ≤9 years and the major parasite was Enterobius vermicularis. Furthermore the overall human infection rate of parasite showed a downward trend with the increase of educational level of the people (χ(2)=70.969, P=0.000 1), the differences had significance (χ(2)=120.118, P=0.000 1). For different populations, the infection rate of intestinal parasites was highest among preschool children. The infection of intestinal helminth was mainly mild, only 2 severe cases were detected. The infection rate of Clonorchis sinensis in urban residents was only 0.006%. Logistic regression analysis showed that being preschool children (χ(2)=15.765, P=0.000 1) and drinking well water (χ(2)=45.589, P=0.000 1) were the risk factors for intestinal parasite infection, and annual income per capita of farmers was the protective factor against intestinal parasite infection. The infection rates of protozoa and intestinal parasites decreased sharply compared with the results of previous two surveys, and the rate of intestinal helminth infection also dropped sharply compared with the second survey. The numbers of protozoa, helminth and intestinal parasites detected in this survey were all less than the numbers found in the previous two surveys. Conclusions: Compared the results of three surveys in Henan, the infection rate of protozoa and intestinal parasites showed a downward trend. The prevention and treatment of Enterobius vermicularis infection in children should be the key point of parasitic disease control in the future.

[Surveillance for viral diarrhea in sentinel hospitals in Henan province, 2013-2015].

Objective: To investigate the infection status of human rotavirus, calicivirus, astrovirus and enteric adenovirus in children aged <5 years in disease surveillance areas in Henan province from 2013 to 2015. Methods: A total of 880 stool samples were collected from four sentinel hospitals and group A rotavirus was detected by ELISA and group A rotavirus G/P genotyping was performed with nested multiplex RT-PCR, while rotavirus (group B, C), calicivirus and astrovirus were detected by two-step multiplex RT-PCR and adenovirus were detected by PCR. The epidemiological data of positive cases were statistically analyzed. Results: A total of 594 positive samples were detected, including 24 mixed infection samples, 370 rotavirus positive samples (42.0%); 162 calicivirus positive samples (18.4%); 69 astrovirus positive samples (7.8%) and 17 enteric adenovirus positive samples (1.9%). The overall positive rate of four viruses was significantly higher in urban area than in rural area, but the positive rate of rotavirus was higher in males than in females and in younger age group than in older age group. G9P [8] was the major genotype of group A rotavirus, there were two seasonal infection peaks in autumn and spring. Norovirus Ⅱ was the predominant type of calicivirus and the infection peak was in spring. Viral diarrhea cases were distributed in different age groups, mainly in age groups 0-12 months (rotavirus) and 3-5 years (calicivirus). The main clinical symptoms included fever, diarrhea and vomiting. The etiological characteristics differed with gender and area. Conclusions: The infection rate of diarrheal viruses was higher in young children <5 years old in disease surveillance areas. The epidemiological and clinical features varied with the type of pathogen.

IL-17 rs2275913 genetic variation contributes to the development of gastric cancer in a Chinese population.

The purpose of this hospital-based case-control study was to assess whether the interleukin (IL)-17 rs2275913 genetic variation can influence susceptibility to gastric cancer. Samples from a total of 202 gastric cancer patients and 237 controls were collected from the Linyi People's Hospital between March 2013 and March 2015. The IL-17 rs2275913 gene polymorphism was identified by polymerase chain reaction and restriction fragment length polymorphism. When compared with control subjects, gastric cancer patients were older in age (OR = 3.89, 95%CI = 2.55-5.95), male (OR = 2.08, 95%CI = 1.39-3.10), had a habit of alcohol consumption (OR = 1.71, 95%CI = 1.15-2.55), and were more likely to be infected with Helicobacter pylori (OR = 2.76, 95%CI = 1.83-4.16). We observed that the AA genotype of the IL-17 rs2275913 polymorphism resulted in a 2.32-fold risk of gastric cancer compared to the GG genotype (OR = 2.32, 95%CI = 1.20-4.54; P = 0.01). The AG combined with AA genotype of the IL-17 rs2275913 polymorphism had more risk of developing gastric cancer than the GG genotype (OR = 1.50, 95%CI = 1.01-2.23; P = 0.04). Moreover, the AA genotype of the IL-17 rs2275913 polymorphism was correlated with a higher risk of developing gastric cancer than the GG and AG genotypes combined (OR = 2.01, 95%CI = 1.08-3.79; P = 0.02). In conclusion, the results of our study suggest that the IL-17 rs2275913 polymorphism could contribute to the risk of gastric cancer.

Predictive potential role of glutathione S-transferases polymorphisms in response to chemotherapy and breast cancer prognosis.

The aim of this study was to evaluate the role of GSTM1 null/present, GSTT1 null/present, and GSTP1 polymorphisms in the clinical response to chemotherapy and treatment outcome of breast cancer. The GSTM1, GSTT1, and GSTP1 IIe105Val polymorphism genotypes were analyzed using polymerase chain reaction coupled with restriction fragment length polymorphism. Conditional logistic regression analysis revealed that breast cancer patients carrying the GG genotype of GSTP1 IIe105Val showed a significantly better response to chemotherapy compared to those expressing the AA genotype [odds ratio = 2.66, 95% confidence interval (CI) = 1.24-5.91, P = 0.007]. The Cox proportional hazards model indicated that the GG genotype of GSTP1 IIe105Val in breast cancer patients was correlated with a lower risk of death from all causes than those with AA genotype. The adjusted hazard ratio (95%CI) for the GG genotype of GSTP1 IIe105Val was 0.44 (0.18-0.99; P = 0.03). In conclusion, the results of our study indicated that the GG genotype of GSTP1 IIe105Val was significantly associated with better response to chemotherapy and longer overall survival, compared to the wide-type genotype.

Effect of estradiol and bisphenol A on human hepatoblastoma cell viability and telomerase activity

Sex hormones from environmental and physiological sources might play a major role in the pathogenesis of hepatoblastoma in children. This study investigated the effects of estradiol and bisphenol A on the proliferation and telomerase activity of human hepatoblastoma HepG2 cells. The cells were divided into 6 treatment groups: control, bisphenol A, estradiol, anti-estrogen ICI 182,780 (hereinafter ICI), bisphenol A+ICI, and estradiol+ICI. Cell proliferation was measured based on average absorbance using the Cell Counting-8 assay. The cell cycle distribution and apoptotic index were determined by flow cytometry. Telomerase activity was detected by polymerase chain reaction and a telomeric repeat amplification protocol assay. A higher cell density was observed in bisphenol A (P<0.01) and estradiol (P<0.05) groups compared with the control group. Cell numbers in S and G2/M phases after treatment for 48 h were higher (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h (P<0.05) were higher in these groups than in the control group. The cell density was also higher in bisphenol A+ICI (P<0.01) and estradiol+ICI (P<0.05) groups compared with the ICI group. Furthermore, cell numbers were increased in S and G2/M phases (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h were higher (P<0.05) in these groups than in the ICI group. Therefore, bisphenol A and estradiol promote HepG2 cell proliferation in vitro by inhibition of apoptosis and stimulation of telomerase activity via an estrogen receptor-dependent pathway.

Effect of estradiol and bisphenol A on human hepatoblastoma cell viability and telomerase activity.

Sex hormones from environmental and physiological sources might play a major role in the pathogenesis of hepatoblastoma in children. This study investigated the effects of estradiol and bisphenol A on the proliferation and telomerase activity of human hepatoblastoma HepG2 cells. The cells were divided into 6 treatment groups: control, bisphenol A, estradiol, anti-estrogen ICI 182,780 (hereinafter ICI), bisphenol A+ICI, and estradiol+ICI. Cell proliferation was measured based on average absorbance using the Cell Counting-8 assay. The cell cycle distribution and apoptotic index were determined by flow cytometry. Telomerase activity was detected by polymerase chain reaction and a telomeric repeat amplification protocol assay. A higher cell density was observed in bisphenol A (P<0.01) and estradiol (P<0.05) groups compared with the control group. Cell numbers in S and G2/M phases after treatment for 48 h were higher (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h (P<0.05) were higher in these groups than in the control group. The cell density was also higher in bisphenol A+ICI (P<0.01) and estradiol+ICI (P<0.05) groups compared with the ICI group. Furthermore, cell numbers were increased in S and G2/M phases (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h were higher (P<0.05) in these groups than in the ICI group. Therefore, bisphenol A and estradiol promote HepG2 cell proliferation in vitro by inhibition of apoptosis and stimulation of telomerase activity via an estrogen receptor-dependent pathway.

Impact of caffeine on ß cell proliferation and apoptosis under the influence of palmitic acid.

We examined the influence of caffeine on the proliferation and apoptosis of ß cells cultured in vitro in the presence of the free fatty acid palmitic acid (PA). Different concentrations of caffeine (1-100 mM) and free fatty PA were added to cultured ß cells. The MTT assay was used to analyze cell proliferative activity; flow cytometry was used to measure apoptosis and calculate the apoptosis rate. Compared with the blank control group, cells cultured with 500 mM PA for 24, 48, 72, and 96 h showed inhibition of pancreatic ß cell proliferative activity. In the 10 and 25 mM caffeine groups cultured for 48, 72, and 96 h, ß cell proliferative activity was much higher than that in the 500 mM PA group. The apoptosis rate in the 500 mM PA group was 40.55 ± 20.33%, which was higher than that in the blank control group. The apoptosis rates in the 10 and 25 mM caffeine group and the PA group were 19.12 ± 10.56 and 20.97 ± 9.75%, respectively, which was lower than that in the 500 mM PA group. At some concentrations, caffeine can improve free fatty PA levels and guide pancreatic ß cell proliferation inhibition and cell apoptosis.

Evaluating the clinical significance of serum HE4 levels in lung cancer and pulmonary tuberculosis.

OBJECTIVE: To evaluate whether serum human epididymis protein 4 (HE4) levels could be used as a marker to differentiate lung cancer from pulmonary tuberculosis (PTB) and as a prognostic indicator in patients with non-small-cell lung cancer (NSCLC). DESIGN: HE4 levels were measured in serum samples from 106 healthy controls, 190 lung cancer patients, 114 patients with PTB and 24 patients with pneumonia using enzyme-linked immunosorbent assay. RESULTS: Serum HE4 levels in lung cancer patients were significantly higher than those in patients with PTB and healthy controls (P < 0.001). Using the optimal cut-off value of 94.01 pmol/l, HE4 levels distinguished lung cancer from PTB with a sensitivity of 61.6% and a specificity of 93.0%. After adjusting for age and smoking status, a binary unconditional logistic regression model provided a sensitivity of 67.4% and a specificity of 86.0% for differentiating between these two diseases. In the NSCLC group, serum HE4 levels were significantly higher in patients at advanced stages (Stage I-II vs. Stage III-IV, P = 0.02). Higher levels of serum HE4 (≥ 83.90 pmol/l) were significantly correlated with a worse 3-year survival rate for patients with NSCLC (P < 0.05). CONCLUSION: Serum HE4 may be used as a potential marker to differentiate lung cancer from PTB and healthy controls. In addition, higher levels of HE4 predict poor prognosis in NSCLC patients.

Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth.

Conventional plasmids for gene therapy produce low-level and short-term gene expression. Here, we first created minicircle carrying endostatin (mc-hES) for measurement of transfection efficiency. Compared with pcDNA-hES, MC-mediated endostatin gene transfer in vitro resulted in seven-fold greater endostatin expression levels in transfected cells and inhibited the growth of Human umbilical vein endothelial cells (HUVEC) more efficiently. HUVEC cell migration and tube-formation assays suggested that MC-mediated endostatin gene has significant anti-migration and anti-tube-formation capacity than that in pcDNA-hES. In vivo experiments showed that after transfection, mc-hES inhibited the growth of nasopharyngeal carcinoma xenografts. The tumor inhibition rates of mc-hES and pcDNA-hES were 60.8% and 26.9%, respectively (P<0.05). MC-mediated intratumoral endostatin expression in vivo was 2.2-17.9 times higher than pcDNA-hES in xenografted mice and lasted for 20 days. Our results suggest that minicircle DNA vectors might be a promising vector for biotherapy and should be further investigated.

[Synthesis of triphenylethylene with aliphatic cyclic moiety and its antagonism on estrogen receptor].

AIM: In order to improve the biological activity and reduce the side effects and toxicity, a series of novel estrogen receptor antagonists were designed. METHODS: The key triphenylethylene intermediates were obtained by the McMurry reaction. The target compounds were prepared by etherification. The binding affinities of the target compounds for the estrogen receptor in rat uterine cytosol were measured by a competitive binding assay and their estrogen agonistic/antagonistic properties were investigated in the 3-day uterine weight assay in the immature rats. RESULTS: Thirty-five new compounds have been synthesized and their geometric configuration were determined by X-ray crystallography and 1HNMR spectral data. CONCLUSION: All of the test compounds showed affinity for the estrogen receptor (IC50 < 10(-6) mol.L-1), especially compound 35 with IC50 1.07 x 10(-8) mol.L-1. Some compounds are antagonists, inhibiting uterus growth; others are agonists, promoting uterus growth. Compounds 14 and 27 are superior antagonists to tamoxifen.