Advances in potential drugs for improvement of energy metabolism in heart failure / 药学实践杂志

Heart failure (HF) is a global public health problem with high morbidity and mortality. Numerous studies have shown that HF is caused by severe disturbance of energy metabolism, resulting in insufficient cardiac energy supply. This lack of energy could lead to a failure of the heart to pump blood and a failure of energy metabolism in other organs throughout the body. Currently, therapeutics of HF work by reducing heart rate and cardiac preload and afterload, symptomatic treatment, or delaying the progression of the disease. However, drugs targeting heart energy metabolism have not been developed. the main characteristics of cardiac energy metabolism, metabolic changes during HF were summarized and drugs that improve cardiac function through energy metabolism were discussed, which could provide a new research direction for the development and application of drugs in treatment of heart failure.

Methodology and application of process analytical technology (PAT) for traditional Chinese medicine manufacturing:a review / 中国中药杂志

Owing to the advancement in pharmaceutical technology, traditional Chinese medicine industry has seen rapid development. Preferring conventional manufacturing mode, pharmaceutical enterprises of traditional Chinese medicine have no effective process detection tools and process control methods. As a result, the quality of the final products mainly depends on testing and the quality is inconsistent in the same batch. Process analytical technology(PAT) for traditional Chinese medicine manufacturing, as one of the key advanced manufacturing techniques, can break through the bottleneck in quality control of medicine manufacturing, thus improving the production efficiency and product quality and reducing the material and energy consumption. It is applicable to the process control and real-time release of advanced manufacturing modes such as intelligent manufacturing and continuous manufacturing. This paper summarized the general idea of PAT for traditional Chinese medicine manufacturing. Through the analysis of the characteristics and status quo of the technology, we summed up the methodology for the continuous application and improvement of PAT during the whole life-cycle of traditional Chinese medicine. The five key procedures(process understanding, process detection, process modeling, process control, and continuous improvement) were summarized, and the application was reviewed. Finally, we proposed suggestions for the technical and regulatory challenges in implementing PAT in traditional Chinese medicine industry. This paper aims to provide a reference for development and application of PAT in advanced manufacturing, intelligent manufacturing, and continuous manufacturing of traditional Chinese medicine industry.

Pharmacokinetic study of HMS-01 in mice / 药学实践杂志

Objective To study the pharmacokinetics of HMS-01 in mice and provide support for subsequent studies. Methods Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to establish a sensitive and specific method for the determination of the concentration of HMS-01 in plasma and other biological samples. The pharmacokinetics of HMS-01 in C57BL/6J mice were studied by the established method. To obtain the basic pharmacokinetic parameters, three doses of HMS-01 were given orally and one dose of HMS-01 was given intravenously. Results The pharmacokinetic results of mice showed that the intestinal absorption of HMS-01 was fast, the oral bioavailability of HMS-01 in mice was moderate (50% to 70%). The exposure levels (AUC and cmax) of HMS-01 in mice increased with the increase of dosage, while the AUC was linearly correlated with the increase of dosage. After intravenous administration of HMS-01, the half-life period in mice was about 1 h which was not long. The plasma clearance rate (CLtotal.p) was 2.8 L/h·kg, which was similar to the hepatic blood flow of mice. The apparent volume of distribution (VSS) was 5 L/kg, which was much larger than the total mouse fluid. There were significant differences in AUC and F (P<0.05), but no significant differences in parameters such as cmax,AUC0−∞,t1/2,CLtot,p,MRT,Vss in male and female mice which were given 30 and 60mg/kg HWS-01 orally. Conclusion The pharmacokinetic process of HMS-01 in mice showed gender differences, and the area under the curve of blood concentration time and bioavailability of female mice were higher than that of male mice. As oral bioavailability was reasonable, further in vivo studies on HMS-01 in mice with heart failure by oral administration could be considered to provide evidence.

Research on batch-to-batch quality control of compound Danshen extract based on physical characterization and multivariate statistical analysis / 中国中药杂志

Physical attributes of Chinese herbal extracts are determined by their chemical components, and the physical and chemical attributes jointly affect the preparation process performance and the final product quality. Therefore, in order to improve the quality control of Chinese herbal extracts, we should comprehensively study the batch-to-batch consistency of physical and chemical attributes as well as the correlations between them. This paper first explored the physical attributes affecting the preparation process performance of the compound Danshen extract and developed a method for characterizing the texture attributes. With such main chemical components as water, phenolic acids, saponins, and saccharides and texture, rheology, and other physical attributes taken into consideration, the batch-to-batch quality fluctuation of products from different production lines and time was analyzed by principal components analysis(PCA). Finally, the correlation and partial least squares(PLS) analysis was conducted, and the regression equation was established. The fitting result of the PLS model for dynamic viscosity was satisfying(R~2Y=0.857, Q~2=0.793), suggesting that the chemical components could be adjusted by the component transfer rate in the extraction process, the impurity removal rate in the alcohol precipitation process, and the water retention rate of the concentration process to meet the control of the extract dynamic viscosity. This study clarified the correlations between physical and chemical attributes of the compound Danshen extract and established a method for controlling its physical attributes based on process regulation, which would provide reference for improving the quality control of Chinese herbal extracts.

Research on feedforward control method of mixing process for compound Danshen Dripping Pills based on quality by design(QbD) / 中国中药杂志

The mixing process is one of the key operation units for solid preparation of traditional Chinese medicine. The physical properties such as particle size, density and viscosity of the mixture are key factors that need to be controlled, which will directly affect the performance of the preparation molding process and product quality. Subsequent dripping process performance and appearance qua-lity of dripping pills will be affected by dynamic viscosity of materials in the mixing process. Based on this, with mixing process of compound Danshen dripping pills as the object, a feedforward control method for the dripping pill mixing process was established based on the concept of quality by design(QbD). Firstly, critical quality attribute(CQA)-dynamic viscosity, critical material attributes(CMAs)-the moisture content of compound Danshen extract, average molecular weight of polyethylene glycol 6000 and critical process parameter(CPP)-mixing temperature were identified through the analysis of properties for multiple batches of the raw materials and excipients as well as technological mechanism. Then the Box-Behnken experimental design was used to establish the regression model among CMA, CPP and CMA(R■=0.972 0, RMSE =16.24) to obtain the design space. Finally, through the verification of three batches within the design space, the mixing process temperature was adjusted according to the properties of the raw materials and exci-pients to achieve accurate control of the dynamic viscosity attribute. The relative deviation between the actual dynamic viscosity value and the target value was less than 3.0 %. The feedforward control of the mixing process of compound Danshen dripping pills was rea-lized in this study, which can contribute to improving quality consistency of the mixing process intermediates, simultaneously provide a reference for the research on the process quality control of other Chinese medicine dripping pills.

Suitable soil moisture contents for water use efficiency and saponins accumulation in Panax notoginseng / 中草药·英文版

Objective: The moisture content in the soil directly affects the yield and quality of Panax notoginseng, especially at the age of three years old. However, the suitable moisture for the growth of P. notoginseng is unknown. In this study, the effects of different soil moisture on the growth of P. notoginseng were studied. Methods: Four different water treatments (0.45 field capacity (FC), 0.60 FC, 0.70 FC, and 0.85 FC) were set up in Shilin County, Yunnan Province, China. The water consumption and daily dynamic of water consumption were determined daily (from April 21 to October 18, 2012), and the daily dynamic of water consumption under different weather conditions (sunny and rainy) was determined. The transpiration coefficient and water use efficiency were calculated through dry matter accumulation and total water consumption. Accumulation of saponins of roots of P. notoginseng were analyzed by HPLC after treated, and the soil moisture content suitable for the growth of P. notoginseng was estimated by regression fitting of the active ingredient accumulation and the soil moisture content. Results: The water consumption of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.89, 3.68, 3.37 and 2.73 kg/plant per day, respectively. The water consumption of P. notoginseng from June to August was greater than other months. The daily dynamic of water consumption on sunny days and sunny days after rain showed a “double peak” feature, and it showed a “single peak” feature on rainy days. The water uses efficiency (WUE) of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.51, 3.32, 4.59, 3.39 gDW/kg H

Mechanisms of Retinoic Acid-AP-1 Signaling Pathway Modulating Secretion of TGF-β2 by Retinal Pigment Epithelial Cells / 中山大学学报(医学科学版)

@#【Objective】To investigate the effects of up-regulating RA signal and inhibiting AP-1 transcriptional activity on TGF-β2 secretion by RPEs and its possible pathways.【Methods】① To investigate the effects of ATRA treat⁃ ment，human retinal pigment epithelial cell line ARPE-19 cells were divided into 5 groups：control group and 4 interven⁃ tion groups（6 h，12 h，24 h and 48 h after RA treatment）. Western blot，RT-qPCR and immunofluorescence staining were carried out to analyze RARβ and c-Fos expression. ②To investigate the effects of RARβ inhibitor LE540 treatment on expression of RARβ and c-Fos that were induced by ATRA，ARPE-19 cells were divided into 4 groups：control group，ATRA group，LE540 group and ATRA+LE540 group. RARβ and c-Fos expression was assessed by western blot and RT-qPCR. ③ To investigate the effects of AP-1 inhibitor T-5224 treatment，ARPE-19 cells were divided into 4 groups：control group and treatment groups（12 h，24 h and 48 h after T-5224 treatment）. EMSA was carried out to ana⁃ lyze the AP-1 DNA binding activity. ④To investigate the effects of LE540 and T-5224 administration on ATRA- induced TGF-β2 secretion，ARPE-19 cells were divided into 4 groups：control group，ATRA group，ATRA+LE540 group and ATRA+LE540 group. Western blot and ELISA were carried out to analyze TGF-β2 secretion in ARPE-19 cells.【Results】 RARβ level in ARPE-19 cells was significantly higher in treatment group than in control group after being treated with ATRA for 24 and 48 hours（P＜0.05）. C-Fos level was first up-regulated and then decreased. After treatment with ATRA for 6 and 12 hours，c-Fos expression were significantly upregulated（P＜0.01），but at 48 h after treatment，their expression were significantly decreased to the level which had no statistical difference compared with the control group （P＞0.05）. The AP-1 DNA binding activity was significantly decreased in ARPE-19 cells after being treated with T-5224 for 24 and 48 hours（P＜0.01）. Compared with ATRA group，TGF-β2 secretion was statistically down-regulated after being treated with LE540 and T-5224 for 48 hours（P＜0.05）.【Conclusion】ATRA can induce TGF-β2 secretion in RPE cells through affecting RARβ expression and AP-1 transcriptional activity.

Investigation of awareness of menopausal hormone therapy among medical staff in Zhujiang Hospital of Southern Medical University / 解放军医学杂志

Objective To investigate the awareness and attitude towards menopause and menopausal hormone therapy (MHT) among the medical staff of Zhujiang Hospital of Southern Medical University.Methods A self-designed questionnaire related to menopause and MHT was conducted among 1143 medical staffs in Zhujiang Hospital of Southern Medical University.Results The best-known symptoms,in sequences,were dysphoria and depression (90.6％),sleep disorders (81.5％),hot flashes and night sweating (69.4％),dizziness and palpitation (59.3％),and paresthesia (50.3％).Of 1143 respondents,42.1％(481) knew about MHT,and 62％(709) considered that MHT is necessary for symptomatic menopausal women.Significant differences existed in attitudes towards MHT between different titles and departments (P=0.027,P=0.000).Fifty-seven percent (651) of medical staff expressed concern about the side effects of MHT and had scruples about its use,73.1％(836) believed that MHT can improve menopausal symptoms,while 54.5％(623) believed MHT can prevent and treat osteoporosis.Conclusions The awareness rate on menopause and MHT is relatively low among the medical personnel of Zhujiang Hospital of Southern Medical University.There exist differences in attitudes towards menopause and MHT among different departments,doctors and nurses,and different titles.

Effect of allogeneic adipose derived mesenchymal stem cells on regulatory T cells in peripheral blood of patients with premature ovarian failure / 解放军医学杂志

Objective To investigate the effect of allogeneic adipose derived mesenchymal stem cells (hADSCs) on regulatory T cells (Tregs) in peripheral blood of patients with premature ovarian failure (POF).Methods hADSCs were isolated from adipose tissue using 0.1％ type Ⅰ collagenase digestion.Adipogenic,osteogenic differentiation and surface molecular characterization were also performed.Peripheral blood mononuclear cells (PBMCs) of POF patients were isolated by density gradient centrifugation.PBMCs were co-cultured with 1 × 104,2 × 104 and 1 × 105 hADSCs for 72 hours under the stimulation of phytohemagglutinin (PHA).The proliferation rate of lymphocytes was measured by CCK-8 method,the proportion of CD4+ CD25+ Foxp3+ Treg cells was measured by flow cytometry.Real-time fluorescent quantitative PCR was used to detect the mRNA level of Foxp3+.Results hADSCs were positive for CD90,CD105 and negative for CD34,CD45,and has adipogenic and osteogenic differentiation ability.The results of CCK-8 showed that hADSCs could significantly inhibit the proliferation of lymphocytes compared with that in control group (P＜0.001).Flow cytometry showed that hADSCs could promote the proliferation of CD4+ CD25+ Foxp3+ Treg cells (P＜0.05).The results of qPCR showed that Foxp3+ mRNA expression was obviously up-regulated in all experimental groups compared with that in control group (P＜0.001).Conclusion hADSCs can play an immune regulatory role and promote the proliferation of Tregs in peripheral blood of patients with premature ovarian failure.

Research on the protective effect of glycyrrhizin against renal ischemia-reperfusion injury in mice and its mechanisms / 中华器官移植杂志

Objective To investigate the protective effect of glycyrrhizin against renal ischemiareperfusion injury in mice and its mechanisms.Methods Male C57BL/6 mice were divided into three groups of six.Bilateral flank incisions were made,the right kidney was removed and the left kidney was subjected to ischemia using a microvascular clamp,which was removed after 30 min.In the shamoperated group,the mice underwent anesthesia,bilateral flank incisions and a right nephrectomy.In the glycyrrhizin-treated group,the mice were injected with 60 mg/ kg glycyrrhizin 1 h prior to ischemia.In the saline-treated group,the mice were administered with 60 mg/ kg saline.The mice were sacrificed 6 h after reperfusion and the blood and kidney samples were immediately collected for kidney function,inflammatory response and signal pathway test.Results As compared with those in the saline-treated group,the mice in glycyrrhizin0-treated group exhibited notably decreased serum levels of creatine and blood urea nitrogen at 6 h following reperfusion (P＜0.01),the SOA level was significantly reduced (P＜0.01) and the SOD activity was increased.The activity of MPO (P＜0.01)in the glycyrrhizin-treated group was significantly reduced as compared with the saline-treated group,also the serum level of pro-inflammatory TNF-α (P＜0.05),IFN-γ (P＜0.05),IL-1β (P＜0.01) and IL-6 (P＜0.01).Furthermore,the phosphorylated-p38 protein level in the glycyrrhizin-treated group was notably as reduced compared with that in the saline-treated group.Conclusion Pretreatment with glycyrrhizin attenuates renal ischemia-reperfusion injury via inhibition of tissue inflammation by downregulating p38 mitogen-activated protein kinase signaling.

Characterization, Quantification and Pharmacokinetic Analysis of Bevacizumab and Its Glycosylation by Mass Spectrometry / 分析化学

The bevacizumab and its glycoforms were analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-MS), sodium dodecyl sulfate polyacrylamide gel electrophoresis ( SDS-PAGE) and short-gun strategy, with the sequence of unique peptide and seventeen glycoforms being characterized. The bevacizumab and its glycopeptides concentrations in mice plasma with different intravenous injection doses of bevacizumab were detected and the concentration-time curves were obtained by parallel reaction monitoring ( PRM) method based on liquid chromatography-tandem mass spectrometry (LC-MS / MS) technique. First, standard curves were created for quantification of mAb in mice plasma, which showed good linearity, with the correlation coefficient (R2 ) value of 0. 998 and the lower limit of quantification of 66 fmol. Detection results of high and low doses of the drug in the mice plasma samples showed that the drug concentration-time curve trend was consistent, e. g. the concentration was decreasing. However, the results of quantitation of seventeen glycoforms demonstrated that the metabolism of different glycoforms was different. The concentrations of most glycoforms increased first, whereas the metabolism afterwards differed by different glycoforms.

Neuroprotective effect of Nogo-66 receptor silencing in preterm rats with brain injury caused by intrauterine infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of Nogo-66 receptor (NgR) silencing with specific small interfering RNA (siRNA) on brain injury repair in preterm rats with brain injury caused by intrauterine infection and related mechanism of action.</p><p><b>METHODS</b>The pregnant Sprague-Dawley rats (with a gestational age of 15 days) were selected, and premature delivery was induced by RU486 or lipopolysaccharide (LPS). The preterm rats delivered by those treated with RU486 were selected as the control group. The preterm rats with brain injury caused by intrauterine infection induced by LPS were divided into model, empty vector, and NgR-siRNA groups, with 36 rats in each group. The rats in the control and model groups were given routine feeding only, and those in the empty vector and NgR-siRNA groups were given an injection of lentiviral empty vector or NgR-siRNA lentivirus via the lateral ventricle on postnatal day 1 (P1) and then fed routinely. On P3, P7, and P14, 8 rats in each group were randomly selected and sacrificed to harvest the brain tissue. RT-PCR was used to measure the mRNA expression of NgR. Western blot was used to to measure the protein expression of active RhoA. The immunofluorescence histochemistry was used to determine the degree of activation of microglial cells and the morphology of oligodendrocyte precursor cells (OPCs). Hematoxylin and eosin staining was used to observe the pathological changes in brain tissue. The behavioral score was evaluated on P30.</p><p><b>RESULTS</b>On P3, the NgR-siRNA group had significantly lower mRNA expression of NgR and protein expression of active RhoA in brain tissue than the model and empty vector groups (P<0.05). In each group, the mRNA expression of NgR was positively correlated with the protein expression of active RhoA (P<0.05). The results of immunofluorescence histochemistry showed that on P3, the NgR-siRNA group had a significantly reduced fluorescence intensity of the microglial cells labeled with CD11b compared with the model and empty vector groups (P<0.05). The OPCs labeled with O4 antibody in the four groups were mainly presented with tripolar cell morphology. The results of pathological examination showed a normal structure of white matter with clear staining in the periventriclar area in the control group, a loose structure of white matter with disorganized fibers and softening lesions in the model and empty vector groups, and a loose structure of white matter with slightly disorganized fibers, slight gliocyte proliferation, and no significant necrotic lesions in the NgR-siRNA group. As for the behavioral score, compared with the model and empty vector groups, the NgR-siRNA group had a higher score in the suspension test, a longer total activity distance, and greater mean velocity and number of squares crossed, as well as a shorter time of slope test and a shorter time and distance of activity in the central area (P<0.05), while there were no significant differences in these parameters between the NgR-siRNA and control groups (P>0.05).</p><p><b>CONCLUSIONS</b>NgR silencing with specific siRNA can effectively silence the expression of NgR in pertem rats with brain injury caused by interauterine infection and has a significant neuroprotective effect in brain injury repair.</p>

Relationship between clinical features and cognitive function in patients with childhood and adolescence-onset schizophrenia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia.</p><p><b>METHODS</b>The clinical data of 78 patients with childhood and adolescence-onset schizophrenia who met with the criteria of ICD-10 for schizophrenia were retrospectively reviewed. The cognitive functions were evaluated by the Chinese Wechsler Intelligence Scale for Children (C-WISC), the Wisconsin Card Sorting Test (WCST), digit span backward and P300. The clinical symptoms were evaluated by the Positive and Negative Syndrome Scale (PANSS).</p><p><b>RESULTS</b>The patients with a lower education level or earlier onset of age had a longer P3 latency at the P300Fz area. The patients with a higher parental education level had higher scores of full intelligence quotient (FIQ), verbal intelligence quotient (VIQ), performance intelligence quotient (PIQ), conceptual level and completed categories of WCST and backward numeric order reciting. The patients with higher PANSS negative subscale scores had lower scores of FIQ, VIQ, PIQ, completed categories and conceptual level of WCST and backward numeric order reciting. The patients with a longer stabilization time had higher backward numeric order reciting scores.</p><p><b>CONCLUSIONS</b>The severity of negative symptoms of the patients and the educational level of their parents are major factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia.</p>

Effect of parent training in combination with methylphenidate treatment on family relationships for children with attention deficit/hyperactivity disorder / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of parent training combined with methylphenidate treatment on family relationships in children with attention deficit/hyperactivity disorder (ADHD).</p><p><b>METHODS</b>Fifty-nine parents of children with ADHD under methylphenidate treatment participated in a modified 5-week training program. The intervention effect was evaluated using the Conners Parent Symptom Questionnaire, ADHD Rating Scale-IV Home Version (ADHD-RS-IV Home Version), Caregiver Strain Questionnaire, Parent-Child Relationship Self-rating Scale and Piers-Harris Children's Self-Concept Scale. Parents also completed the training satisfaction survey before and after the intervention.</p><p><b>RESULTS</b>After the 5-week parent training, compared with the baseline values, total scores of Conners Parent Symptom Questionnaire and scores of conduct problems and anxiety significantly decreased, and scores of attention deficit, hyperactivity, impulsivity and oppositional defiant behaviors of ADHD-RS-IV Home Version, and Caregiver Strain Questionnaire total scores were all significantly decreased (P<0.05), while total scores of the Parent-Child Relationship Self-Rating Scale and Piers-Harris Children's Self-Concept Scale were significantly increased (P<0.05).</p><p><b>CONCLUSIONS</b>Modified 5-week parent training program may improve parent-child relationship and reduce parenting stress in ADHD families.</p>

Study on attention-deficit hyperactivity disorder in children based on resting-state functional magnetic resonance imaging / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To compare resting-state functional magnetic resonance imaging (fMRI) findings of children with attention-deficit hyperactivity disorder (ADHD) and normal children, and to investigate the possible mechanism of brain dysfunction in children with ADHD.</p><p><b>METHODS</b>Resting-state fMRI was performed on 18 children who met the DSM-IV diagnostic criteria for ADHD (ADHD group) and 18 normal children (control group) matched for age, sex, IQ, degree of education and handedness. The two groups were compared in terms of amplitude of low frequency fluctuation (ALFF) and regional homogeneity (ReHo).</p><p><b>RESULTS</b>Compared with the control group, the ADHD group had decreased ALFF in the bilateral posterior lobes of the cerebellum and the left side of the pons, increased ALFF in the right precentral gyrus, decreased ReHo in the left medial frontal gyrus, right superior frontal gyrus, and left precuneus, and increased ReHo in the left anterior lobe of the cerebellum, left caudate nucleus, right parahippocampal gyrus, left precentral gyrus, and right middle frontal gyrus.</p><p><b>CONCLUSIONS</b>In resting state, children with ADHD have decreased brain activity in some regions, including the cerebellum and frontal cortex, compared with normal children, which supports the hypothesis of dysfunctional fronto-cerebellar circuits in ADHD.</p>

Influential factors for methylphenidate treatment compliance in children with attention deficit hyperactivity disorder / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate influential factors for the tendency to medicate and medication compliance in children with attention deficit hyperactivity disorder (ADHD).</p><p><b>METHODS</b>A total of 188 children aged from 5 to 16 years, who were initially diagnosed with ADHD according to DSM-IV criteria, were included in the study. They underwent symptom assessment and cognitive function test. The compliance of methylphenidate treatment was evaluated.</p><p><b>RESULTS</b>Patients with better emotional state, and fewer oppositional and hyperactive behaviors and those who had a family history of psychiatric diseases and who obtained lower scores in the number cancellation test (NCT), were more prone to medication and/or exhibited better medication compliance. Logistic regression analysis showed that fewer oppositional and hyperactive behaviors and lower NCT scores were the predictive factors for a higher tendency to medicate, and a better emotional state was the predictive factor for better medication compliance. Patients of predominantly inattentive type were more prone to medication and showed better medication compliance, as compared with those of combined type. Gender, age and symptom severity were not associated with the tendency to medicate and/or medication compliance.</p><p><b>CONCLUSIONS</b>There is a need to enhance medication compliance in children with ADHD who have hyperactive, impulsive and oppositional behaviors, and to improve their long-term social functions.</p>

Effects of hypoxia-indudble factor 1α on the growth of subcutaneously implanted hepatocellular carcinoma in nude mice / 中华普通外科杂志

Objective To study the effects of HIF-1α expression on the growth of subcutaneously implanted human hepatocellular carcinoma (HCC) cell lines in nude mice. Methods A nude mouse model of subcutaneously implanted HCC cell line HepG2Tet-on-HIF-1α was established. This cell lines characterizes the inducible expression of HIF-1α by doxycycline (Dos). The impact of HIF-1α expression induced by Dox on the growth of subcutaneously implanted HCC cell lines in the nude mouse model was observed. Results The mRNA and protein expression of HIF-1α were significantly up-regulated in the nude mouse model by oral administration of Dox. Compared with that model in which Dox was not administrated ie Dos( - ) group, the tumor volume(513.545 ± 276. 229) mm3 vs. ( 166. 506 ± 110. 142) mm3 ( P < 0. 05 ), tumor weight ( 1.251 ± 0. 438 ) g vs. (0. 640 ± 0. 296) g ( P < 0. 05 ), and tumor growth velocity were significantly enhanced in Dox ( + ) group, while tumor necrosis was inhibited ( 31. 360% ±2. 728% vs. 36. 640% ± 3. 804% ) (P<0. 05). The weight loss of nude mice was larger in Dox( + )group( P < 0. 01 ). There was no liver or lung metastasis in either group. Conclusion The expression of HIF-1αin subcutaneously implanted HCC in a nude mouse model is up-regulated by oral Dox. High grade expression of HIF-1α promotes the growth of implanted HCC.

Expressions of ATP binding cassette transporter genes in rat hepatic oval cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Liver regeneration occurs through hepatocytes after acute liver injury. However, severe liver injury activates bipotential oval cells from canals of Hering which can differentiate into hepatocytes and biliary epithelial cells. Most models of oval cell activation have employed potential carcinogens to inhibit hepatocyte replication in the face of a regenerative stimulus. Oval cells must be able to withstand the toxic milieu of the damaged liver. ATP binding cassette transporters are cytoprotective efflux pumps that may contribute to the protection of these cells. The aim of this study was to determine the ABC transporter expressions in hepatic oval cells.</p><p><b>METHODS</b>A rat model was established by feeding 2-acetylaminofluorene combined with partial hepatectomy to activate hepatic oval cells. Oval cells were isolated and purified using selective enzymatic digestion and density gradient centrifugation from the heterogeneous hepatic cell population. The expressions of ABC transporter gene, including MDR1, MRP1 and Bcrp1, in isolated hepatic oval cells and hepatocytes were measured by quantitative real-time reverse transcription-polymerase chain reaction and those in rat liver tissues were measured by immunohistochemistry.</p><p><b>RESULTS</b>Compared to those in the rat hepatocytes, mRNA expressions of the genes encoding MDR1, MRP1 and Bcrp1 were increased up to 9-, 1.5- and 13.8-folds in hepatic oval cells. Immunohistochemical staining of rat liver slides demonstrated that the expression of MDR1 proteins was found around periportal areas, and Bcrp1 protein was found located on cell membranes.</p><p><b>CONCLUSION</b>Hepatic oval cells express high levels of the ABC transporter gene that may have cytoprotective functions during severe hepatotoxicity.</p>

Study on sensitivity to chemotherapy by combination therapy with tumor necrosis factor-alpha and bromocriptine in the multidrug resistant subcell line of HepG2 / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the change of chemosensitivity of hepatocarcinoma cell line (HepG(2)/ADM) after treated by bromocriptine (BCT) combination with human tumor necrosis factor-alpha (TNF-alpha).</p><p><b>METHODS</b>Firstly, TNF-alpha gene was transfected into HepG(2)/ADM cell line by liposome to establish a cell model expressing the TNF-alpha protein stably. All experiments were divided into four groups and named blank control group (group A), drug resistant group HepG(2)/ADM (group B), TNF-alpha gene group HepG(2)/ADM/TNF (group C) and BCT group (group D) respectively. And group D came from group C treated with BCT simultaneously. MTT assay was tested to detect the sensitivity to ADM of each group and Rhodamine 123 (Rh123) was applied to test the function of P-gp by flow cytometric analysis (FCM). MDR associated genes and proteins and PKC-alpha protein were detected by immunohistochemistry (IHC), Western blot and reverse transcriptase polymerase chain reaction (RT-PCR) methods, respectively. The expression and the apoptosis rate of Bcl-2 in the hepatocarcinoma cells were detected by FCM.</p><p><b>RESULTS</b>There was significant difference between group C and D in the rate of reversing resistance and the intracellular Rho123 accumulation (P < 0.01). MDR1 mRNA and P-gp protein expression in group C and D were low similar to that in group A, but no difference could be found among them (P > 0.05). As we found that PKC-alpha protein expression was downregulated in group D but Bcl-2 protein expression was downregulated in group C, and there was significant difference compared to other groups. The apoptosis rate of hepatocarcinoma cells was much higher in group D than that in group C (P < 0.01) with FCM, but similar to group A (P > 0.05).</p><p><b>CONCLUSIONS</b>Synergistic effect of BCT and TNF-alpha on reversing hepatocellular carcinoma multidrug resistance could enhance the susceptibility of HepG(2)/ADM cells to cytotoxic drugs.</p>

Inhibition of the VEGF Expression and Cell Growth in Hepatocellular Carcinoma by Blocking HIF-1α and Smad3 Binding Site in VEGF Promoter / 华中科技大学学报（医学）（英德文版）

In order to investigate the inhibitory effects on the vascular endothelial growth factor (VEGF) expression and cell growth in hapatocellular carcinoma (HCC) by blocking HIF-1α and Smad3 binding site in the VEGF promoter, antisense oligodeoxynucleotides (ASODN) were designed to block HIF-1α and Smad3 binding site in the VEGF promoter. Different concentrations of ASODN and ODN were transfected into HCC cells respectively. The expression of VEGF mRNA and protein was detected by SABC, Western blot and RT-PCR techniques and the inhibitory effects on the expression of VEGF and cell growth of the HCC cells stimulated by the supernatants were determined by using MTT method. Immunohistochestry revealed that after co-inoculation of hepatocellular carcinoma cells with different concentrations of ODN and ASODN for 48 h, there was no significant difference in the expression of VEGF protein between ODN group and control group (P＞0.05), but there was significant difference between ASODN group and control group (P＜0.05). At a concentration of 10 μmol/L ASODN, the difference was very significant (P＜0.01).Western blot and RT-PCR revealed that, after treatment for 48 h at a concentration of 10 μmol/L,the integral gray levels and RNA odds were 59743.2±10412.5 and 0. 783±0. 032 in ODN group,and 38694.5±10925. 1 and 0. 468±0. 015 in ASODN group, respectively, with the difference being very significant (P＜0.01). Antisense ODN could inhibit the growth of HCC cells in a concentration-dependent manner. It was concluded that anti-gene technique of aiming at HIF-1α action site in the VEGF promoter could suppress the VEGF expression and inhibit HCC cell growth, and it is promising that anti-gene technique works as a new gene therapeutic tool for anti-angiogenesis of HCC.