Objective To elucidates the effects of HPV18 E6 siRNA targeting at human papillomavirus(HPV)18 E6 gene on the proliferative activity of HeLa cells and chemotherapy sensitivity.Methods HPV18 E6 expression of HeLa cells was inhibited by siRNA interference,the change of P53 and P21 proteins expression level was measured by Western blot.MTT assay was used to detected proliferative activity and sensitivity to paclitaxel liposome of HeLa cells.Results After inhibition of E6 expression,P53 and P21 proteins increased and the growth of HeLa cells was decreased(P ＜0.01).The inhibition rate of HeLa was markedly increased after transfection of HPV18 E6 siRNA and paclitaxel liposome.Conclusion HPV18 E6 siRNA can effectively silence gene expression of E6 and inhibit proliferation of HeLa cells.HeLa cells are more sensitive to combine HPV18 E6 siRNA with paclitaxel liposome than that of control groups.

Objective To analyze the clinical features and factors affecting prognosis for intracerebral hemorrhage (ICH) during pregnancy and postpartum.Methods A study of ICH was performed on 61 women in Beijing Tiantan Hospital,Capital Medical University between January 1997 and December 2014,and all cases were diagnosed with cerebral hemorrhage or subarachnoid hemorrhage during pregnancy or six weeks after delivery with CT or MRI after exclusion of ICH due to craniocerebral trauma.The subjects were divided into surgery (n=26) and conservative treatment (n=35) groups according to different ways of treatment;pregnancy associated problems (n=11) and cerebrovascular diseases groups according to the aetiology of ICH;low (n=13) and high score group (n=48) according their Glasgow score at the first visit;and short clinical onset to diagnosis time (O-D time) group (≤ 24 h,n=33) and long O-D time (＞24 h) group (n=28).We compared the maternal clinical features and prognosis between different groups with t,Mann-Whitney U or Chi-square tests.A stratified logistic regression was used to assess the effect of factors affecting the prognosis.Results The average gestational age at the onset of ICH of the 61 cases was (28.8±8.3) weeks (6-40 weeks),the Glasgow score was (11.3±4.8),the median O-D time was 24.0 h,the modified Rankin scale (mRS) was 2.7,and 14 maternal deaths were reported (23.0％).Among the 61 women,three were terminated in early trimester,12 terminated in second trimester,and the rest 46 delivered in late term among which two fetal deaths,44 live births,and four neonatal deaths.Thus the perinatal infant death rate was 13.0％ (6/46).The difference of maternal clinical features and prognosis between the surgery and conservative treatment group was not significant (all P＞0.05).However,comparison between the cerebrovascular disease and pregnancy associated diseases group showed the latter had a lower Glasgow score and Apgar score [12.2(3.0) vs 7.5(12.0),(8.9±1.9) vs (7.2±2.6)],the higher mRS [2.4(2.0) vs 3.9(5.0)] and gestational age [(27.7±8.4) vs (34.9±4.1)],maternal mortality rate [14.0％ (7/50) vs 7/11] and perinatal death rate [5.4％ (2/37) vs 4/11] (t or x2=-3.09,-2.34,1.93,1.17,2.12 and 1.78,all P＜0.05).For women with low Glasgow score,the median O-D time was shorter than that of higer Glasgow score group (8.0 vs 48.0 h),the mRs and maternal mortality rate were higher 4.9(2.5) vs 3.1(2.0);9/13 vs 10.4％(5/48),t,U or x2=426.00,5.77 and 19.14,all P＜0.05].The short O-D time group showed lower Glasgow score and average Apgar score of the newborns than the long O-D time group [9.8(11.3) vs 13.2(2.0),(7.9±2.7) vs (9.2±0.9);t,U or x2=-2.91 and-2.23,both P＜0.05].The Glasgow scores was negatively associated with the mRs (OR=-0.26,95％CI:-0.16 to-0.05).In particular,O-D time (OR=0.03,95％CI:0.00-0.66) and pre-eclampsia (OR=0.33,95％CI:0.12-0.26) were both positively related to maternal mRs.However,the Glasgow scores,surgical treatment,O-D time and concomitant pre-eclampsia were irrelevant to the death ofperinatal infants (all P＞0.05).Conclusions The prognosis is poor in women with ICH during pregnancy or postnatal period whose Glasgow score was low or O-D time was long,or the ICH occurred due to pre-eclampsia.Antenatal care should be strengthened and early identification and diagnosis might improve the prognosis.

Objective: Farnesyl pyrophosphate synthase (FPS) gene of Fritillaria thunbergii was cloned and the correlation between the expression content of gene and alkaloid accumulation was analyzed to explore the role of FPS gene in the regulation of alkaloid synthesis and metabolism. Methods: In this study, the new bulbs and different tissues of Fritillaria thunbergii varieties "Narrow leaf" "Broad leaf" and "New Meiyuan" were used to clone the full-length sequence of FPS gene via RACE technology. RT-qPCR and HPLC-ELSD technology was used to determine the expression of FPS in 10 different developmental stages and the content of alkaloid (Peimine and Verticinone), respectively. Finally, the correlation was analyzed. Results: The full-length cDNA sequence of FPS gene was 1 629 bp. Open Reading Frame (ORF) was 1 056 bp and encoded 351 amino acids. Among them, the tissue specific expression trends of FPS gene were same, flowers had the highest expression level, followed by the new bulb. The results of correlation analysis showed thatthe content Peimine and Verticinone had significant positive correlation with the expression level of FPS gene in "Broad leaf" and "New Meiyuan" (The correlation coefficients were 0.289 and 0.613, 0.427 and 0.622, respectively);FPS gene expression and alkaloid content in different tissues had a positive correlation (The correlation coefficients were 0.057 and 0.476, 0.085 and 0.495, 0.375 and 0.432, respectively). Conclusion: The full-length sequence of FPS gene was successfully cloned in this study. The FPS gene involved in the regulation of alkaloid synthesis and metabolism.

Objective To detect the expression of IL-17 in renal transplant recipients by Luminex and evaluate the relationship between the level of serum IL 17 and early acute renal allograft rejection.Methods 38 kidney transplant recipients and healthy controls (HC,healthy volunteers,n =20) were selected in this study from January 2009 to October 2011.All patients were divided into two groups according to their allograft outcome as acute rejection group (ARG,n-18) and non-rejection group (NRG,n=20).The expression of serum IL-17 was detected by Luminex technique in two groups of kidney transplant recipients and HC.To evaluate the correlation between the level of serum IL-17 and early acute renal allograft rejection.Results The mean level of IL-17 in all renal transplant recipients 1.3 ± 1.9 pg/ml at the pre-transplantation was significantly lower than that in HC (6.9± 8.5 pg/ml,t=3.968,P＜0.001).The results highlighted that the level of serum IL-17 in ARG 3.4±5.8 pg/ml was significantly higher than that in NRG (0.5±0.4 pg/ml) on the 7th days post Kidney transplantation (post KTx,t=2.242,P =0.031).Kaplan-Meier survival analysis showed that the probability of rejectionfree survival in the higher levels group of IL-17 on the 7th days post-KTx was significant lower than that in the lower levels group (P＜0.001).The results of receiver operating characteristic curve showed that the sensitivity and specificity of the combined IL-17 to predict acute rejection were 66.67％ and 100％,respectively.Conclusion The serum IL-17 levels in renal transplant recipients on the 4th and 7th post-KTx days can predict early renal transplant rejection.

Objective To develop an online organ doses reporting software VirtualDose-IR, which can compute the radiation doses and provide an easy access to evaluation and control of patients ′radiation doses.Methods Monte Carlo method was applied to simulating various interventional radiology ( IR) processes , which included various parameters such as different patient models at different ages and with different weights , different projection angles and regions of interest , and other parameters .All of the dose data was acquired and then integrated into a database , and displayed with hyper text markup language (HTML), so only a web browser was necessary for users .Results A web-based software that reports organ doses for patients under IR progress was developed .The organ doses assessed with VirtualDose-IR were compared with other experiment and simulation data , and the results were basically consistent with each other .Conclusions VirtualDose-IR is a easy and efficient method to assess patients′radiation doses of IR.

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

Objective To observe the effect of Yiqi-Yangyin-Quyu decoction combined with acupuncture on serum homocystine level and blood coagulation function of patients with cerebral infarction in convalescent period. Methods A total of 80 patients with cerebral infarction convalescence were divided into the observation group and the control group according to the random number table method, with 40 cases in each group. The control group was given routine treatment of western medicine. Based on this, the observation group was treated with Yiqi-Yangyin-Quyu decoction combined with acupuncture. After 4 weeks of treatment, the Barthel index (BI), neurological impairment National Institutes of Health Stroke Scale (NIHSS), C-reactive protein (CRP), and the level of homocystine (HCY) and blood coagulation function index of plasma prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (FIB)] of the two groups were compared and analyzed. Results The total effective rate of the observation group was 90.0％ (36/40), while the total effective rate of the control group was 70.0％ (28/40). The total effective rate of the 2 groups was statistically significant (Z=-3.184, P=0.004). After treatment, the NIHSS score of the observation group were lower than those of the control group,and the BI score of the observation group were higher than those of the control group(t value were 4.528, 4.337, P<0.05). After treatment, the content of CRP (9.25 ± 1.21 mg/L vs. 15.24 ± 1.74 mg/L, t=4.905) and HCY (12.59 ± 2.05 μmol/L vs. 16.52 ± 2.05 μmol/L, t=4.821) of the observation group were significantly lower than those of the control group (P<0.05). After treatment, the level of PT (15.20 ± 1.17 s vs. 12.95 ± 1.02 s, t=4.891), APTT (36.72 ± 1.98 s vs. 33.02 ± 1.68 s, t=4.553), and TT (14.67 ± 0.95 s vs. 12.21 ± 1.10 s, t=4.210) of the observation group were significantly higher than those of the control group (P<0.05), while the level of FIB (2.55 ± 0.25 g/L vs. 3.03 ± 0.51 g/L, t=4.027) of the observation group was significantly lower than this of the control group (P<0.05). Conclusions The application of Yiqi-Yangyin-Quyu decoction combined with acupuncture can improve the blood coagulation function, reduce the injury of brain tissue, promote the recovery of cerebral nerve function, and improve the quality of life of the patients.

Objective To investigate the effect of compound Fufangteng mixture-containing serum on the proliferation of bone marrow mesenchymal stem cell (BMSC) and its mechanism. Methods Rat BMSC were isolated, cultured and purified in vitro by direct adherence method. Cell morphology was observed. Surface markers were identified by flow cytometry. The rats were treated with compound Fufangteng mixture at a dose of 3 mL/(kg·d) by gavage for 14 d, and then the drug-containing serum was collected. BMSC were divided into the blank control group, drug-containing serum group, Notch1 small interfering ribonucleic acid (siRNA) group and Notch1 siRNA+drug-containing serum group. The proliferation rate of BMSC was detected and the relative expression levels of Notch1 signaling pathway-associated messenger ribonucleic acid (mRNA) and proteins were measured in each group. Results Microscopic observation showed that the first generation BMSC were seen in the long spindle shape, and grown in the parallel or spiral pattern. The third generation BMSC positively expressed CD90 and CD44, whereas were negative for CD45. Compared with the blank control group, the proliferation rate of BMSC in the drug-containing serum group and Notch1 siRNA+ drug-containing serum group was significantly increased, whereas that of BMSC was significantly decreased in the Notch1 siRNA group (all P < 0.05). Compared with the Notch1 siRNA group, the proliferation rate of BMSC was significantly increased in the Notch1 siRNA+drug-containing serum group (P < 0.05). Compared with the blank control group, the relative expression levels of Hey1 and Delta-like ligand (DLL)1 mRNA and proteins were significantly up-regulated in the drug-containing serum group, whereas those were significantly down-regulated in the Notch1 siRNA group and Notch1 siRNA+drug-containing serum group (all P < 0.05). Compared with the Notch1 siRNA group, the relative expression levels of Hey1 and DLL1 mRNA and proteins were significantly up-regulated in the Notch1 siRNA+drug-containing serum group (all P < 0.05). Conclusions Compound Fufangteng mixture-containing serum may promote the proliferation of rat BMSC, and its mechanism is probably associated with the activation of Notch1 signaling pathway.

Objective To evaluate the effect of mesenchymal stem cell (MSC) coated-islets on instant blood-mediated inflammatory reaction (IBMIR) after islet transplantation. Methods MSC labeled with tracer and human islets were placed into an ultra-low adsorption cell culture dish, shaken and mixed twice at an interval of 0.5 h, and then incubated at 37 ℃ and 5% CO₂ for 24 h to obtain MSC-coated islets. The coating effect of MSC and in vitro function of the islets were assessed. A blood circulation tube-shaped model was established in vitro. In the blank control group, 0.2 mL of islet culture solution was added. In the islet group, 800 islet equivalent quantity (IEQ) of uncoated islets were supplemented. In the MSC-coated islets group, 800 IEQ of MSC-coated islets were added, and circulated for 60 min at 37 ℃. A portion of 0.5 mL blood sample was taken for routine blood test at 0, 30 and 60 min, respectively. After 60 min circulation, the blood sample was filtered with a 70 μm filter to collect plasma, blood clots and islets. Blood clots and islets were subject to hematoxylin-eosin (HE) staining and immunohistochemical staining. Morphological changes and the aggregation of CD11b-positive cells surrounding the islets were observed. The contents of plasma thrombin-antithrombin complex (TAT), tissue factor (TF), C3a, C5b-9, interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1 and IL-8 were determined by enzyme-linked immune absorbent assay. Results After 24 h co-incubation, the islets were coated by MSC, with a coating degree of approximately 80%. In the islet and MSC-coated islet group, a large quantity of neutrophils and monocytes were observed surrounding the blood clots and islets, and the quantity of CD11b-positive cells in the MSC-coated islet group was less compared with that in the islet group. After co-incubation with the whole blood for 0, 30 and 60 min, the quantity of platelets, neutrophils and monocytes was declined in the MSC-coated and islet groups, and gradually decreased over time. Compared with the blank control group, the quantity of platelets, monocytes and neutrophils was lower, whereas the TF content was higher in the MSC-coated islet group. Compared with the islet group, the quantity of platelets, monocytes and neutrophils was higher, whereas the TAT and TF contents were less in the MSC-coated islet group, the differences were statistically significant (all P < 0.05). Compared with the blank control group, the expression levels of C3a, C5b-9, IL-6, TNF-α and IL-8 were up-regulated in the MSC-coated islet group. Compared with the islet group, the expression levels of C3a, C5b-9, IL-1β, IL-6, TNF-α, IL-8 and MCP-1 were down-regulated in the MSC-coated islet group, and the differences were statistically significant (all P < 0.05). Conclusions MSC-coated islets may reduce the exposure of islet TF in the blood and prevent the incidence of IBMIR during the coagulation response stage, thereby mitigating the injury and loss of islet allograft in the early stage of islet transplantation.

Islet transplantation is one of the effective therapies for diabetes mellitus. Nevertheless, multiple issues still exist, such as shortage of donors and adverse reactions caused by long-term use of immunosuppressants, which limit the islet survival post-transplantation. Microencapsulated islet transplantation may overcome these difficulties to certain extent, whereas many factors, such as the destruction of immune isolation microenvironment within the microcapsules and insufficient supply of oxygen and nutrients, constrain the application of microencapsulated islet transplantation in clinical practice. In recent years, how to enhance the effect of microencapsulated islet transplantation has been gradually studied. The application of stem cells in microencapsulated islet transplantation has steadily become a research hot spot. Therefore, the optimizing strategies for microencapsulated islet transplantation and the application of stem cells in microencapsulated islet transplantation were reviewed, and the potential improvement techniques of microencapsulated islet transplantation were investigated in this article, aiming to provide reference for further clinical application of microencapsulated islet transplantation.