The aim of this study was to study the synthesis of two folate conjugates and their application in the preparation of folate targeted liposome, and to investigate their targeting effect in hepatocellular carcinoma HepG2 cell line in vitro. In this study, Folate-PEG-Cholesteryl hemisuccinate(Folate-PEG2000-CHEMS and Folate-PEG4000-CHEMS)were synthesized by linking folate and cholesterol succinate with two kinds of PEG materials. Structures of Folate-PEG2000-CHEMS and Folate-PEG4000-CHEMS were characterized by 1H NMR and ultra-high resolution mass spectrometry. Calcein was selected as the model drug, and calcein liposomes FA-PEG2000-L and FA-PEG4000-L were prepared by film dispersion method using Folate-PEG2000-CHEMS and Folate-PEG4000-CHEMS, respectively. The particle size and Zeta potential of FA-PEG2000-L and FA-PEG4000-L were measured by laser particle size analyzer. The drug delivery effect of FA-PEG2000-L and FA-PEG4000-L was evaluated by cellular uptake experiment in HepG2 cell line in vitro. Flow cytometry and laser confocal scanning microscope were used to determine fluorescence in HepG2 cells in vitro. The results showed that the average particle size of calcein liposome was (205.8 ± 10.2) nm, and the Zeta potential of calcein liposome was -(1.19 ± 0.31) mV.There was no significant difference in particle size and Zeta potential between FA-PEG2000-L and FA-PEG4000-L. The fluorescence intensity of FA-PEG4000-L liposome group was about 3.6 and 3.1 times higher than that of non-targeted liposome group and FA-PEG2000-L liposome group, with statistically significant difference (P < 0.01). The drug delivery efficiency of FA-PEG4000-L group in HepG2 cells was higher than that in FA-PEG2000-L and non-targeted groups, and the results indicated that Folate-PEG4000-CHEMS can promote the uptake of liposome by HepG2 cells in vitro. All in all, Folate-PEG4000-CHEMS could be applied in the preparation of folate targeted liposome, which could promote the uptake of liposome by HepG2 cells.

AIM:To explore the effects of miR-21 on biological behavior of colon cancer cells and their sensi-tivity to epidermal growth factor receptor monoclonal antibody cetuximab .METHODS:Lentiviral vectors were constructed to generate up-and down-regulations of miR-21 lentiviruses (LV-miR-21 and LV-anti-miR-21, respectively), and the cor-responding negative control viruses (LV-miR-21 NC and LV-anti-miR-21 NC, respectively) were also constructed.The vi-ruses were used to infect human colon cancer RKO cells .The changes of the miR-21 expression level , the cell prolifera-tion, the colony-forming ability, the cell apoptosis and the sensitivity of the cells to cetuximab were detected by real -time PCR, MTT assay, soft agar colony assay , flow cytometry and CCK-8 assay.RESULTS: The lentivirus titers of LV-miR-21, LV-miR-2 NC, LV-anti-miR-21 and LV-anti-miR-21 NC were 3.0 ×1012 TU/L, 6.0 ×1011 TU/L, 2.0 ×1012 TU/L and 8.0 ×1011 TU/L, respectively.The infection efficiency was over 80% by the observation of green fluorescence .The miR-21 expression level , the cell proliferation , and the colony-forming ability in LV-miR-21 group were significantly higher than those in LV-anti-miR-21 group.The early apoptotic rate and the inhibitory rate of cetuximab for the cells in LV-anti-miR-21 group were higher than those in LV-miR-21 group.CONCLUSION: miR-21 promotes the proliferation of colon cancer cells.Down-regulation of miR-21 enhances the sensitivity of the colon cancer cells to the targeted therapy drug cetuximab.

This study aimed to investigate whether β-elemene could improve the dysfunction of vascular endothelial cells induced by low shear force (LSS), and the proliferation and migration of vascular smooth muscle cells induced by oxidized low-density lipoprotein (ox-LDL). Parallel plate flow chambers and ox-LDL were used to establish vascular endothelial cells (ECs) dysfunction model and vascular smooth muscle cell (VSMCs) proliferation and migration model, respectively, and the effects of β-elemene on ECs dysfunction and VSMCs proliferation and migration were examined. The activity of ROS in ECs was measured by DHE and the activity of NO in ECs was tested by DAF-FM DA. The protein phosphorylation of Akt and ERK in ECs were detected by Western blot. The proliferation of VSMCs was measured by MTT. The migration of VSMCs was examined by cell scratch test and Transwell assay. The gene expression of MMP-2 and MMP-9 in VSMCs was measured by RT-qPCR. In ECs, β-elemene could significantly reduce the LSS-induced increase in ROS, significantly increase the LSS-induced decrease in NO, decrease the phosphorylation of ERK, and increase the phosphorylation of Akt. In VSMCs, β-elemene could significantly reduce the proliferation and migration of VSMCs induced by ox-LDL, and reduce the gene expression of MMP-2 and MMP-9. To conclude, β-elemene can improve the LSS-induced ECs dysfunction and ox-LDL-induced VSMCs proliferation and migration.

In this study, the conjugate of eicosapentaenoic acid (EPA) and hyaluronic acid (HA) was synthesized and the anti-hepatoma activities in vitro were evaluated.The hyaluronic acid-eicosapentaenoic acid (HA-EPA)nanoparticle was synthesized by linking eicosapentaenoic acid with hyaluronic acid with cystamine.The structure of HA-EPA was characterized by nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FT-IR).Laser particle sizer and Zeta potential analyzer were used to detect the size and potential of HA-EPA.MTT assay was used to detect the anti-proliferative effect of HA-EPA on HepG2, Huh-7 and LX-2 cells in vitro.The effects of HA-EPA nanoparticles on the proliferation and apoptosis of HepG2 cells in vitro were investigated by EdU staining and TUNEL staining. The apoptosis was further confirmed by flow cytometry.The effect of HA-EPA nanoparticles on the migration and invasion of HepG2 cells was demonstrated by transwell and invasion experiments.The results of 1H NMR showed that HA-EPA was successfully synthesized, and the grafting rate of EPA on HA was (40 ± 5) %. The structure of HA-EPA was further confirmed by FT-IR.The particle size was (162.5 ± 10.2) nm, and the potential was -(4.47 ± 0.31) mV.MTT results showed that, with the prolongation of drug treatment time, HA-EPA showed a better inhibitory effect on the activity of HepG2 and Huh-7 cells than EPA under the same EPA content.After treated for 48 hours, the toxicity of HA-EPA to LX-2 cells was less than that of EPA.The results of 24-hour proliferation, apoptosis, migration and invasion of HepG2 showed that, the graft of hyaluronic acid improved the ability of EPA to inhibit proliferation, promote apoptosis, migration and invasion of HepG2 cells (P < 0.001), indicating that grafting of HA can significantly enhance the inhibitory effect of EPA on liver cancer with some role in reducing toxicity.

Radiation-induced bystander effect (RIBE) refers to that irradiated cells release signaling factors and induce responses in nonirradiated cells.In other words, it is the communication between irradiated and nonirradiated cells by intracellular signals. RIBE could influence the efficacy of tumor radiotherapy, but also has potential risk to the normal tissues outside of radiation field. Studies have found that ionizing radiation can induce the alteration of miRNA expression not only in the irradiated cells but also in adjacent nonirradiated tissues, and miRNAs may play an important role in the regulation of signaling pathways between irradiated and nonirradiated bystander cells. This article reviewed the roles of miRNAs in RIBE.

OBJECTIVE: To detect VHL gene mutation in a pedigree affected with von Hippel-Lindau syndrome (VHL). METHODS: Clinical data of the pedigree was reviewed. Patients were subjected to Sanger sequencing to detect mutation of the VHL gene. Structure of pVHL was predicted by 3D modeling using the swiss-model. RESULTS: A novel c.426delT(p.V142fs) [NM_000551] mutation was found in exon 2 of the VHL gene. 3D modeling suggested that the alpha-structure of pVHL is completely absent. CONCLUSION The novel c.426delT(p.V142fs) mutation probably underlies the VHL in this pedigree.
DNA Mutational Analysis ; Exons ; Humans ; Mutation ; Pedigree ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics ; von Hippel-Lindau Disease ; genetics

Objective: To investigate the prevalence of calf muscle cramps and possible influencing factors for pregnant women in China. Methods: Using a multi-stage stratified probability proportional to size cluster randomization sampling method during 2010-2012. A total of 3 582 pregnant women were investigated at 150 counties from 31 provinces in China mainland. Information on calf muscle cramps, demographic socio-economic status, pregnancy information, and the physical activities was collected through questionnaires. The semi-quantitative food frequency questionnaire (FFQ) was used to collect food intake of pregnant women. Dynamic cluster analysis was used to assess dietary pattern. Multiple logistic regression was used to investigate the possible influencing factors for calf muscle cramps. Results: The prevalence of calf muscle cramps was 32.9% (1 180/3 582) in Chinese pregnant women, which was 11.6% (87/748), 28.2% (420/1 492), and 50.2% (673/1 342), respectively, during the first, second and third trimester. There were significant differences between them (χ²=349.16, P<0.001). Dietary patterns of the pregnant women were classified into three groups, which called relatively balanced pattern, high vegetables and fruits pattern, and high dairy pattern. Among the three groups, the prevalence of calf muscle cramps was 32.0% (952/2 971), 37.2% (186/500), and 37.8% (42/111), with significant differences (χ²=6.39, P=0.041). The OR (95%CI) values of calf muscle cramps in the second and third trimester was 2.96 (2.28-3.83), and 8.02 (6.16-10.44), respectively, comparing with the first trimester. The OR (95%CI) values of calf muscle cramps in the women taking calcium before pregnant was 1.45 (1.19-1.76), comparing with the one who was not taken. The OR (95%CI) values of calf muscle cramps in the women who had been diagnosed by pregnancy-induced hypertension was 5.76 (2.06-16.12), comparing with the one who had not been diagnosed. The OR (95%CI) values of calf muscle cramps in the high vegetables and fruits pattern and high dairy pattern was 1.13 (0.91-1.41), and 1.18 (0.76-1.81), respectively, comparing with the relatively balanced pattern. Conclusion The prevalence of calf muscle cramps was relatively high in Chinese pregnant women, which was significantly different among three trimesters. The residential areas, occupation, and pregnancy-induced hypertension might be related to the prevalence of calf muscle cramps. However, there was no significant difference among different dietary patterns.

Objective: To investigate the prevalence of anemia and related risk factors for pregnant women in China. Method: Based on Chinese National Nutrition and Health Surveillance 2010-2012, a total of 3 501 pregnant women were investigated from 150 counties of 31 provinces, municipalities and autonomous regions in China, using a multi-stage stratified cluster randomization sampling method. General information of pregnant women, health status, and food intake during the gestation, was collected through a questionnaire investigation. 6 ml fasting venous blood was collected for the determination of hemoglobin concentration. Data were analyzed using multiple logistic regression to investigate the prevalence of anemia and the related influencing factors. Results: The 605 of 3 501 pregnant women had anemia. The prevalence of anemia was 17.2%; and mild anemia accounted for about 61.0% (369/605). Compared with the pregnant women living in the large cities, the OR (95%CI) of those living in the poor rural areas was 1.46 (1.08-1.98). Compared with the pregnant women living in the south area of China, the OR (95%CI) of those living in the north area of China was 1.39 (1.15-1.68); Compared with the pregnant women in the first trimester, the OR (95%CI) of those in the second trimester and the third trimester were 1.79 (1.33-2.43) and 2.11 (1.56-2.85), respectively. The OR (95%CI) of pregnant women who had used folic acid supplementation within the 6 months prior to gestation was 0.76 (0.63-0.93) compared with those who had not used. Conclusion From 2010 to 2012, the epidemic characteristics of anemia was mild for the pregnant women in China, and pregnant women residential areas, periods of pregnancy and whether to take folic acid were related to anemia.

Objective: To examine the status and related factors for gestational weight gain of Chinese pregnant women at different trimesters in 2010-2012. Methods: Participants were from Chinese National Nutrition and Health Surveillance in 2010-2012. Using a multi-stage stratified cluster random sampling method, we recruited 2 805 singleton pregnant women with gestational age 13 weeks or more from 31 provinces of China. A standard questionnaire was used to collect general information and pre-pregnancy weight; body weight and height of pregnant women were measured using a unified weighing scale and stadiometer, dietary intake during the previous year was collected using a food frequency questionnaire. A multiple logistic regression was used to analyze potential factors associated with appropriate gestational weight gain. Results: Among 2 805 pregnant women, 1 441 were in the second (13-27 weeks) and 1 364 in the third trimesters (≥28 weeks) . In the 2^nd trimester, 229 cases (15.9%), 440 cases (30.5%) and 772 cases (53.6%) were insufficient gestational weight gain, appropriate gestational weight gain and excessive gestational weight gain respectively. So were 256 cases (18.8%), 474 cases (34.8%), 634 cases (46.5%) in the 3^rd trimester respectively. In the multivariate unconditional logistic model, less fruit intake was associated with insufficient weight gain for women in the 2^nd trimester (OR (95%CI): 1.58 (1.06-2.34)). Pregnant women with Han ethnicity who live in the small/medium city had lower risk of insufficient weight gain in the 3^rd trimester (OR (95%CI): 0.58 (0.34-0.98)). Those pregnant women with physical activity <1 hour/day had a higher risk of excessive weight gain (OR (95%CI): 1.33 (1.02-1.73)). Conclusion The prevalence of appropriate gestational weight gain was low in China. Our study suggests that pregnant weight gain is associated with fruit intake, types of residential area and physical activity.

Objective: To analyze the status of gestational weight gain (GWG) among Chinese mothers who gave singleton and full-term births, and to look at optimal GWG ranges. Methods: In 2013, using the multi-stage stratified and population proportional cluster sampling method, we investigated 8 323 mother-child pairs at their 0-24 months postpartum from 55 counties (cities/districts) of 30 provinces (except Tibet) in mainland China. Questionnaire was used to collect data on body weight before pregnancy and delivery, diseases during gestation, hemorrhage or not at postpartum, child birth weight and length, and other information about pregnant outcomes. We measured mother's body weight and height, and child's body weight and length. Based on 'Chinese Adult Body Weight Standard’, we divided mothers into four groups according to their body weight before pregnancy: low weight (BMI<18.5 kg/m²), normal weight (BMI 18.5-23.9 kg/m²), overweight (BMI 24.0-27.9 kg/m²) and obesity (BMI≥28.0 kg/m²). The status of GWG was assessed by IOM optimal GWG guidelines. Chinese optimal GWG ranges were calculated according to the association of GWG with pregnant outcomes and anthropometry of mothers and children, and according to P25-P75 of GWG among mothers who had good pregnant outcomes and good anthropometry, and whose children had good anthropometry. The status of GWG was assessed by the new optimal ranges. Results: P50 (P25-P75) of GWG among the 8 323 mothers was 15.0 (10.0-19.0) kg. According to the proposed optimal GWG ranges of IOM, the proportions of inadequate, optimal and excessive GWG accounted for 27.2% (2 263 mothers), 36.2% (3 016 mothers) and 36.6% (3 044 mothers). The optimal GWG ranges for low weight, normal weight, overweight and obesity were 11.5-18.0, 10.0-15.0, 8.0-14.0 and 5.0-11.5 kg. Based on these optimal GWG ranges established in this study, the rates of inadequate, optimal and excessive GWG were 15.7% (1 303 mothers), 45.0% (3 744 mothers) and 39.3% (3 276 mothers), and these rates were significantly different from that defined by the IOM standards (χ2=345.36, P<0.001). Conclusion The median of GWG among Chinese mothers is 15.0 kg, which is at a relatively higher level. This study suggests the optimal GWG ranges for Chinese women who give singleton and full-term babies, which appears lower than IOM's.