An ultra-sensitive and easy-to-use assay for sensing human UGT1A1 activities in biological systems / 药物分析学报

The human UDP-glucuronosyltransferase 1A1 (UGT1A1), one of the most essential conjugative enzymes, is responsible for the metabolism and detoxification of bilirubin and other endogenous substances, as well as many different xenobiotic compounds. Deciphering UGT1A1 relevance to human diseases and characterizing the effects of small molecules on the activities of UGT1A1 requires reliable tools for probing the function of this key enzyme in complex biological matrices. Herein, an easy-to-use assay for highly-selective and sensitive monitoring of UGT1A1 activities in various biological matrices, using liquid chromatography with fluorescence detection (LC-FD), has been developed and validated. The newly developed LC-FD based assay has been confirmed in terms of sensitivity, specificity, precision, quanti-tative linear range and stability. One of its main advantages is lowering the limits of detection and quantification by about 100-fold in comparison to the previous assay that used the same probe substrate, enabling reliable quantification of lower amounts of active enzyme than any other method. The precision test demonstrated that both intra- and inter-day variations for this assay were less than 5.5％. Further-more, the newly developed assay has also been successfully used to screen and characterize the regu-latory effects of small molecules on the expression level of UGT1A1 in living cells. Overall, an easy-to-use LC-FD based assay has been developed for ultra-sensitive UGT1A1 activities measurements in various biological systems, providing an inexpensive and practical approach for exploring the role of UGT1A1 in human diseases, interactions with xenobiotics, and characterization modulatory effects of small mole-cules on this conjugative enzyme.

Pancreatic lipase inhibitory constituents from Fructus Psoraleae / 中国天然药物

Pancreatic lipase (PL), a crucial enzyme in the digestive system of mammals, has been proven as a therapeutic target to prevent and treat obesity. The purpose of this study is to evaluate and characterize the PL inhibition activities of the major constituents from Fructus Psoraleae (FP), one of the most frequently used Chinese herbs with lipid-lowering activity. To this end, a total of eleven major constituents isolated from Fructus Psoraleae have been obtained and their inhibition potentials against PL have been assayed by a fluorescence-based assay. Among all tested compounds, isobavachalcone, bavachalcone and corylifol A displayed strong inhibition on PL (IC < 10 μmol·L). Inhibition kinetic analyses demonstrated that isobavachalcone, bavachalcone and corylifol A acted as mixed inhibitors against PL-mediated 4-methylumbelliferyl oleate (4-MUO) hydrolysis, with the K values of 1.61, 3.77 and 10.16 μmol·L, respectively. Furthermore, docking simulations indicated that two chalcones (isobavachalcone and bavachalcone) could interact with the key residues located in the catalytic cavity of PL via hydrogen binding and hydrophobic interactions. Collectively, these finding provided solid evidence to support that Fructus Psoraleae contained bioactive compounds with lipid-lowering effects via targeting PL, and also suggested that the chalcones in Fructus Psoraleae could be used as ideal leading compounds to develop novel PL inhibitors.

Male external genitalia growth curves and charts for children and adolescents aged 0 to 17 years in Chongqing, China / 亚洲男科学杂志(英文版)

Genital size is a crucial index for the assessment of male sexual development, as abnormal penile or testicular size may be the earliest visible clinical manifestation of some diseases. However, there is a lack of data regarding penile and testicular size measurements for Chinese boys at all stages of childhood and puberty. This cross-sectional study aimed to develop appropriate growth curves and charts for male external genitalia among children and adolescents aged 0-17 years in Chongqing, China. A total of 2974 boys were enrolled in the present study. Penile length was measured using a rigid ruler, penile diameter was measured using a pachymeter, and testicular volume was determined using a Prader orchidometer. Age-specific percentile curves for penile length, penile diameter, and testicular volume were drawn using the generalized additive models for location, scale, and shape. Very similar growth curves were found for both penile length and penile diameter. Both of them gradually rose to 10 years of age and then sharply increased from 11 to 15 years of age. However, testicular volume changed little before the age of 10 years. This study contributes to the literature covering age-specific growth curve and charts about male external genitalia in Chinese children and adolescents. These age-related values are valuable in evaluating the growth and development status of male external genitalia and could be helpful in diagnosing genital disorders.

Research progress on role of macrophages in tumor immunotherapy / 中国免疫学杂志

Objective:Accumulating evidences prove that macrophages affect tumor initiation and prognosis. Especially,tumor-associated macrophages (TAMs) are considered to enhance tumor growth,progression,angiogenesis,invasion and metastasis. Furthermore,tumor suppressive microenvironment can " reeducate" and polarize the phenotype and function of TAM. Recently, researchers use different ways to interfere and modify the phenotype and function of macrophages,to improve the recognition, phagocytosis and antigen presentation of macrophage,and enhance its antitumor activity. We will discuss the pro-tumorigenic properties of macrophage and TAM-targeting therapy as a promising novel strategy for tumor immunotherapy.

Penis growth and development in children and adolescents: a study based on GAMLSS / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate penis development in children and adolescents aged 0-16 years, and to plot the percentile curve for penis development in different age groups.</p><p><b>METHODS</b>A total of 3 024 normal male neonates, children, and adolescents aged 0-16 years in Chongqing, China were selected by simple random sampling and stratified cluster sampling. The length and diameter of the penis were measured for all subjects. A descriptive statistical analysis was used to investigate the data characteristics of the penis, and the GAMLSS fitting model was used to plot the percentile curves of P, P, P, P, P, P, and P97 and obtain percentile reference values.</p><p><b>RESULTS</b>The length and diameter of the penis grew rapidly before the age of 1 year, grew relatively slowly from 1 to 11 years old, and entered a rapid growth period from 11 years old. The length of the penis was positively correlated with its diameter (r=0.961, P<0.01). The percentile reference values of penis length and diameter were obtained and the percentile curve was plotted.</p><p><b>CONCLUSIONS</b>The growth and development of penis length is consistent with that of penis diameter in male children and adolescents in Chongqing, and 0-1 year and 11-16 years are rapid growth periods of penis length and diameter. The percentile curve of penis length and diameter in children and adolescents aged 0-16 years in Chongqing which has been established will provide a reference for further studies on sexual development in children and adolescents.</p>

Diagnostic analysis for 52 cases of gastrointestinal neuroendocrine tumor / 中南大学学报(医学版)

OBJECTIVE@#To investigate the clinical features, diagnosis, treatments and prognosis for gastrointestinal neuroendocrine tumors (GI-NETs).
@*METHODS@#Clincal data of 52 patients, who were diagnosed as GI-NETs between January 2004 and October 2014, were reviewed. The patients were divided into a local excision group (n=21) and a transabdominal excision group (n=30), and the major clinical features, treatment modalities and outcomes were analyzed.
@*RESULTS@#The clinical features of GI-NETs were nonspecific, and most of the clinical manifestation were local invasiveness. CT scan was lack of specific findings. GI-NETs greater than 1 cm often showed local incrassation, upheaval and soft tissue shadow. In the case of lager GI-NETs, necrosis and moderate enhancement could be seen. Positive ratio for expression of chromogranin A (CgA) and synaptophysin (Syn) in the 52 cases of specimen were 63.5% and 88.5%, respectively. Except 1 patient, whose surgery was canceled because of poor health, other 51 patients were treated with surgery through different approaches. Among them, 30 cases were transabdominal resection (57.7%) and 21 were local resection (40.4%). Chemotherapy and/or radiotherapy was only applied for 7 patients. After a follow-up of 40 (3-132) months, 7 patients died, the rest were alive. The median survival in the local resection group and the transabdominal resection group was 43.0 and 39.5 months, respectively (P>0.05).
@*CONCLUSION@#Under the condition of fully understanding the biological characteristics of GI-NETs, early diagnosis and timely personalized treatment is hopeful to reach the relative good prognosis and survival.

Effect of concurrent chemoradiotherapy for serum matrix metalloproteinase 2 and transforming growth factor β1 in patients with locally advanced non-small cell lung cancer and its significance / 肿瘤研究与临床

Objective To compare concurrent chemoradiotherapy and sequential therapy effect on serum MMP-2 and TGF-β1 in local advanced non-small cell lung cancer (NSCLC).Methods From 2010 January to 2012 December,64 Ⅱ B and Ⅲ B stage patients with pathologically confirmed NSCLC were randomly divided into concurrent chemoradiotherapy group (group A) and sequential therapy group (group B).Each group had 32 patients.Group A was treated with three-dimensional conformal radiotherapy and concurrent chemotherapy with TC or EP.Group B received TC or EP regimen chemotherapy after three-dimensional conformal radiotherapy.Serum MMP-2 and TGF-β1 on those patients from preradiotherapy,radiotherapy in one month to post-treatment were measured by enzyme-linked immunoabsorbent assay.The dynamic changes of MMP-2 and TGF-β1 were compared.Results The remission rates in groups A and B were 90.6 ％ and 68.8 ％,the effective rate of treatment in group A was better than that of group B (x2 =4.730 0,P =0.029 6).The long-term effect analyzed with Kaplan-Meier method,the median time to tumor progression (TTP) were 9.1 months and 8.2 months,there was no statistically significant difference (P =0.100 3).The overall survival rates between two groups after the Log-rank test had significant difference (P =0.048),the median survival time (MST) were 17.8 and 15.9 months,1 year OS rates were 65.05 ％ and 60.24 ％,2 years OS rates were 49.45 ％ and 43.07 ％.The MMP-2 level of A group and B were (276.5±98.2) μg/ml and (263.9±103.5) μg/ml,there was no significant difference (t =0.499 6,P =0.619 1) before radiotherapy,they were (242.1±53.2) μg/ml and (298.7±68.4) μg/ml after radiotherapy,there was significant difference (t =3.694 9,P =0.005) and after treatment were (60.5 ±24.4) μg/ml and (75.2±30.7) μg/ml,there was significant difference (t =2.120 5,P =0.038 0).The TGF-β1 level of A group and B were (1 624.3±454.2) ng/ml and (1 564.9±517.8) ng/ml,there was no significant difference (t =0.208 6,P =0.835 4) before radiotherapy,they were (1 383.5±469.3) ng/ml and (1 785.3±412.6) mg/ml after radiotherapy,there was significant difference (t =3.637 3,P =0.006 0) and after treatment were (610.5±215.4) ng/ml and (750.3±263.7) ng/ml,there was significant difference (t =2.322 6,P =0.023 5).Conclusions Concurrent chemoradiotherapy could effectively antagonize radiation-induced MMP-2 and TGF-β1 expression increased in locally advanced NSCLC.This study suggests that the concurrent chemoradiotherapy can inhibit abilities of tumor invasion and metastasis through decreasing the MMP-2 and TGF-β1 levels.

RNA interference targeting c-Met inhibits proliferation of human laryngeal carcinoma Hep-2 cell line in vitro and in its xenografts in nude mice / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>The proto-oncogene c-Met was found to express on human laryngeal carcinoma Hep-2 cell line in previous research. In the present study, the author further examined whether inhibition of c-Met by RNA interference (RNAi) might inhibit biologic activity of Hep-2 cell line in vitro and proliferation using a murine laryngeal carcinoma model.</p><p><b>METHODS</b>RNAi plasmid that can express small interfering RNA targeting c-Met or siRNA that did not match any known human coding mRNA(control siRNA plasmid)was designed, constructed, and transfected into Hep-2 cell line by using cationic liposome Lipofectamine2000 as transfecting agent. In vitro, the transfection efficacy was tested by RT-PCR and Western Blot method, then elected the most inhibitive c-Met-siRNA sequence. Cell proliferation, movement and invasion were studied using MTT, cell migration assay and cell invasion assay, respectively. The Hep-2 cells were transplanted into nude mice, then the time of tumor formation and growth were observed. After tumor formation, c-Met-siRNA was given as the anti-tumor therapy. Expression of c-Met, MMP-9 and VEGF were detected by Western Blot method.</p><p><b>RESULTS</b>After the pSilencer2.0/c-Met-shRNA recombinant plasmid transfection into laryngeal carcinoma Hep-2 cells, the expression of mRNA and protein of c-Met decreased significantly in Hep-2 cells. On the 35th day after tumor vaccination, the tumor volume was (138 ± 27) mm³ in c-Met-siRNA transfection group, Which was diminished significantly in contrast with control group (P < 0.01). The expression of c-Met, MMP-9 and VEGF in the tumor of experiment group was decreased significantly, respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>The results indicated that c-Met-siRNA can down-regulate the expression of c-Met and markedly inhibit laryngeal carcinoma Hep-2 cell proliferation, movement and invasion and the growth of transplantation tumor of nude mice. The siRNA expressing plasmid mediated gene therapy might be a new strategy in targeting molecular therapy of cancer of larynx.</p>

Effects of c-Met-siRNA on the biological behaviour of laryngeal carcinoma Hep-2 cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the effects of c-Met-siRNA on the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells in vitro.@*METHOD@#Firstly, the pSilencer 2.0/c-Met-shRNA recombinant plasmid was transfected into laryngeal carcinoma Hep-2 cells with transfecting agent of cationic liposome Lipofectamine 2000. Secondly,the transfection efficacy was tested by RT-PCR and Western-Blot, then the most inhibitive c-Met-siRNA sequence was elected. Cell proliferation, movement and invasion were detected with MTT, cell migration assay and cell invasion assay, respectively.@*RESULT@#After the transfection of pSilencer 2.0/c-Met-shRNA recombinant plasmid into laryngeal carcinoma Hep-2 cells, the expression of mRNA and protein of c-Met decreased significantly in Hep-2 cells, and ability of the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells were also inhibited.@*CONCLUSION@#The results indicated that c-Met-siRNA can down-regulated the expression of c-Met and markedly inhibited laryngeal carcinoma Hep-2 cell proliferation, movement and invasion. It may have the potential as a therapeutic modality to treat human laryngeal carcinoma.

Action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells in vitro.</p><p><b>METHODS</b>We cultured the Sertoli cells isolated from rat testicular tissues for 2 days in vitro, divided them into a control group incubated with DMSO and 3 case groups exposed to p,p'-DDE and / or beta-BHC at the final concentration of 10, 30, 50 micromol/L for 24 hours, and then detected the expression levels of JNK and c-jun mRNA by two-step RT-PCR.</p><p><b>RESULTS</b>Twenty-four hours after p,p'-DDE treatment, the grayscale values of JNK mRNA were 0.068 +/- 0.001, 0.164 +/- 0.002, 0.207 +/- 0.006 and 0.499 +/- 0.017, and those of c-jun mRNA were 0.122 +/- 0.002, 0.157 +/- 0.006, 0.218 +/- 0.007 and 0.289 +/- 0.004 respectively in the DMSO control and the 10, 30 and 50 micromol/L groups. The expressions of JNK and c-jun mRNA were elevated with increased concentration of p,p'-DDE, with significant differences between the control and the case groups (P < 0.05), and they were also significantly upregulated in the beta-BHC and p,p'-DDE + beta-BHC groups in a dose-dependent manner. The grayscale values of JNK mRNA in the p,p'-DDE, beta-BHC and p,p'-DDE + beta-BHC groups at the concentration of 10 micromol/L were 0.164 +/- 0.002, 0.149 +/- 0.003 and 0.178 +/- 0.004, and those of c-jun mRNA were 0.157 +/- 0.006, 0.131 +/- 0.004 and 0.172 +/- 0.002, respectively, both significantly higher in the combination group than in the former two (P < 0.05). And the same was the case with the 30 and 50 micromol/L concentrations.</p><p><b>CONCLUSION</b>Both p,p'-DDE and beta-BHC can enhance the expressions of JNK and c-jun in Sertoli cells, and their combination can produce even more obvious effect.</p>

Reduced Incidence and Severity of Collagen-induced Arthritis in Mice Lacking LFA-1 / 中国生物工程杂志

Lymphocyte function associated antigen-1 (LFA-1) is a member of integrin family, that plays an important role in the adhesion of lymphocytes with other cells and matrix. To investigate the role of LFA-1 in collagen-induced arthritis (CIA), the incidence of CIA, histological and radiological assessments in the LFA-1 deficient (LFA-1~ -/- ) mice and control mice were examined. LFA-1~ -/- mice and control mice were immunized with 100?g collagen type II(CII) emulsified with an equal volume of Freund’s complete adjuvant (CFA), followed by the booster injection of the same amount of CII in CFA on day 21. Then, clinical, histological and radiological assessments were done. It showed that 57% control mice developed arthritis and apparently changed in the histological and radiological assessment, whereas the all of LFA-1~ -/- mice had the normal histological and radiographic response and none developed arthritis. These results suggeste that LFA-1 is indispensable for the onset of CIA.

The inhibitory action of the antisense oligodeoxynucleotide to the expression of vascular endothelial growth factor by radiotherapy in a prostate cancer cell line / 中华男科学杂志

<p><b>OBJECTIVES</b>To investigate mechanism for the increasing level of serum vascular endothelial growth factor(VEGF) in tumour patients during radiotherapy and the inhibitory action of the antisense oligodeoxynucleotide (AS-ODN) to the expression of VEGF protein by radiotherapy in the prostate cancer cell line (PC3M).</p><p><b>METHODS</b>To observe the changes of serum VEGF in the prostate cancer patients during radiotherapy dynamically and the inhibitory action of the antisense oligodeoxynucleotide to the expression of VEGF by radiotherapy in PC3M.</p><p><b>RESULTS</b>The changes of serum VEGF in three patients receiving radiotherapy had been observed continuously. The levels of serum VEGF began to increase when the patients received radiotherapy and rised up to peak value after fifteen days, then declined to the range of pre-radiotherapy. Irradiating the PC3M cells with X-rays significantly increased the VEGF expression and secretion. The expression of VEGF protein in the group treated by VEGF AS-ODNs and X-ray irradiation decreased significantly than the group treated only by X-ray irradiation.</p><p><b>CONCLUSIONS</b>The induction of VEGF protein expression by X-ray irradiation in tumor cells may result in the increasing of the VEGF in the prostate cancer patients during radiotherapy and the induction can be blocked by VEGF AS-ODNs.</p>