Complex genetic variation has been known to occur during the transmission of human enterovirus (HEV), and the HEV virulence and pathogenicity enhanced by genetic recombination also pose a serious threat to human health. In recent years, the interest in recombination mechanism of genetic plasticity has been renewed with the emergence of pathogenic recombinant circulating vaccine-derived polioviruses, which were implicated in poliomyelitis outbreaks in several regions of the world with insufficient vaccination coverage. This paper reviews recent research progress in HEV genome, including evolutionary characteristics, recombination types, and in vitro recombinant construction.
Animals ; Biomedical Research ; trends ; Enterovirus ; classification ; genetics ; Enterovirus Infections ; virology ; Humans ; Recombination, Genetic ; Viral Proteins ; genetics

Objective: To observe the effects of electroacupuncture (EA) on the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment (CI), and to explore the mechanism of EA in improving the learning and memory abilities. Methods: A total of 100 Sprague-Dawley (SD) rats were divided into a normal group (n=10) and a model group (n=90) by random number table method. Rats in the model group were intraperitoneally injected with a small dose of streptozotocin (STZ) to establish the type 2 diabetic models, after being fed with high-fat and high-sugar diet for 1 month. Twenty CI rats were selected from the 50 successful model rats by the Morris water maze (MWM) test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data (n=10). Rats in the EA group received acupuncture at Zusanli (ST 36), Neiting (ST 44) and Yishu (Extra), of which Zusanli (ST 36) and Neiting (ST 44) were stimulated by EA apparatus, 20 min/time, once a day for 6 d a week and 4 consecutive weeks. The rats in the model and the normal groups were fixed without treatment. After 4-week treatment, the random blood glucose level of the rats was measured; the learning and memory abilities of rats were measured by MWM; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptotic cells; Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex. Results: After modeling, the random blood glucose level and the escape latency tested by MWM were significantly increased, and the number of crossing the platform tested by the MWM was decreased in the EA and model groups, and were significantly different from those in the normal group (P<0.05 or P<0.01), while the differences between the model group and the EA group were not statistically significant (all P>0.05). After 4-week treatment, the random glucose level and the escape latency tested by MWM were significantly increased (both P<0.05), and the number of crossing the original platform tested by the MWM was significantly reduced (P<0.01), the protein and gene expressions of Bax and Caspase-3 were significantly increased (all P<0.001), the protein and gene expressions of Bcl-2 were significantly reduced (both P<0.001), and the number of neuron apoptosis was significantly increased (P<0.001) in the model group than in the normal group; the random blood glucose level was significantly reduced (P<0.05), the escape latency tested by MWM was significantly shortened (P<0.05), and the number of crossing the original platform tested by MWM was significantly increased (P<0.05), the protein and gene expressions of Bax and Caspase-3 were significantly reduced (all P<0.001), the protein and gene expressions of Bcl-2 were significantly increased (both P<0.001), and the number of neuron apoptosis was significantly reduced (P<0.001) in the EA group than in the model group. Conclusion: EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI; the action mechanism may be achieved via anti-apoptosis.

OBJECTIVETo study the proliferation effect of the AM supernatant incubated activation of p38 mitogen activated protein kinases (p38MAPK) signal transduction pathway in human embryonic lung fibroblasts, and to participate in the development of fibrosis in silicosis.

METHODSThe silicotic alveolar macrophages were collected by bronchoalveolar lavage and incubated in vitro in the DMEM medium containing SiO₂ (50 µg/ml) and DMEM medium without SiO₂ for 18 h. Then the AM supernatant incubated for 18 h was collected. HELFs were isolated by organize paste block method, and incubated with AM supernatants. HELFs were divided into four groups: blank control groups, AM groups, SiO₂ + AM groups, SB203580 + SiO₂ + AM groups. The proliferation in the HELF was detected with MTT method and Flow cytometry.

RESULTSThe proliferation in the HELF acted with the conditioned AM supernatant fluid were more than blank control groups, AM groups and SB203580 + SiO₂ + AM groups [average optical density: (0.48 ± 0.03) vs (0.29 ± 0.01), (0.38 ± 0.02), (0.33 ± 0.03)], the values with MTT method were statistically different (P < 0.05); Proliferous index with flow cytometry in SiO₂ + AM groups (18.12 ± 0.82) was bigger than blank control groups (9.24 ± 0.48), AM groups (14.76 ± 0.43) and SB203580 + SiO₂ + AM groups (11.71 ± 0.70) and the values were statistically different(P < 0.05).

CONCLUSIONSThe AM supernatant stimulated by silicon dioxide can accelerate the proliferation in the HELF by activation of p38MAPK signal transduction pathway.

Adult ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; Fibroblasts ; cytology ; drug effects ; pathology ; Humans ; Lung ; cytology ; MAP Kinase Signaling System ; Macrophages, Alveolar ; cytology ; Male ; Signal Transduction ; Silicon Dioxide ; pharmacology ; Silicosis ; metabolism ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To investigate the relationship of pathological response of breast cancer after neoadjuvant chemotherapy with the imaging findings in dynamic contrast-enhanced MRI.Methods Forty- five patients with pathologically confirmed breast carcinoma who finished courses of neoadjuvant chemotherapy had breast MRI prior to operation.Dynamic contrast-enhanced MRI scans were performed on a 1.5 T scanner using 3D SPGR sequence before and repeated 6 times after administration of Gd-DTPA. Pathological response was assessed by a pathologist according to Miller & Payne five points classification blinded to breast MRI results.Grade 5 was defined as pCR(pathological complete response).Grade 4 and 5 were defined as major histopathological response(MHR).The type of time signal intensity curve(TIC) (three types),pattern of residual enhancement of each breast cancer were recorded and correlated with pathological findings.Fisher exact test was used for statistical analysis.Results Grade 5 responses were achieved in seven patients;grade 4 in sixteen patients;grade 3 in sixteen patients and grade 1—2 in six patients.70.0%(14/20)of type Ⅰ time signal intensity curve correlated with MHR,while all 6 type Ⅲ curves showed non-MHR response.The type of time signal intensity curve and pathological response grades had statistically significant correlation(P=0.001).18 of the 23 cases with MHR exhibited residual enhancement,while the remaining 5 cases showed no enhancement.Of the 18 MHR cases with residual enhancement,11 showed non-mass-like enhancement and 7 showed mass-like enhancement.The mass(non- mass)morphological pattern in dynamic contrast enhanced-MRI had statistically significant differences in pathological response(P=0.012).Conclusions Pathological response of breast carcinoma after neoadjuvant chemotherapy could be characterized using dynamic contrast-enhanced MRI by identifying patterns of residual contrast enhancement and kinetic curve.Favorable pathological responses correlated with Type Ⅰ TIC,non-enhancement,and non-mass-like residual enhancement.

Growth year is one of the important factors for the quality of Polygala tenufolia. In this study, primary metabolites and secondary metabolites were compared in 1, 2 and 3 years old P. tenufolia cultivated in Shaanxi Heyang. The samples were subjected to ultra-high performance liquid chromatography (UPLC)-quadrupole time-of-flight mass spectrometry (Q-TOF MS) and nuclear magnetic resonance (NMR) analysis, and the obtained data were analyzed using principal component analysis (PCA) and other statistical analysis methods. In addition, content and correlation of different metabolites were also calculated. The results showed no significance between main component contents in 2 year-old and 3 year-old P. Tenufolia, but 1 year-old was statistically different. The contents of primary metabolites, such as fructose, sucrose, and choline increased as time goes on, while glycine and raffinose decreased. The contents of secondary metabolites, such as onjisaponin Fg, polygalasaponin XXVIII, polygalasaponin XXXII increased, while polygalaxanthone III and parts of oligosaccharide multi-ester including tenuifoliose A, tenuifoliose C, tenuifoliose C2 and tenuifoliose H decreased with the extension of the growth years. Growth years has important impact on the quality of P. tenuifolia and the existing growing years of commodity P. tenuifolia have its scientific evidence. This study supplied a new method for the quality evaluation of Chinese medicinal materials.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Metabolomics ; Oligosaccharides ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Quality Control

Objective To explore the biological exposure limit of liver dysfunction induced by arsenic-coal burning, and screen sensitive biornarkers for its' liver dysfunction monitoring. Methods One hundred and eighteen subjects from the exposed area and 50 control from non-pollution area were studied. Their urinary and hair contents of arsenic were tested as exposure biomarkers by Ag-DDC assay. Total bile acid(TBA, detected by enzymatic cycling method), glutathione S-transferase (GSTs, detected by chemical colorimetry) and γ-glutamyl transpeptidase (γ-GT, detected by colorimetry of diazotization reagent) were used as biomarkers indicating liver cell damage. were used as liver fibrosis biomarkers. The benchmark dose (BMD) and the lower confidence limit of benchmark dose(BMDL) of urinary and hair arsenic were calculated. Sensitivity of each biomarker was estimated according to the BMD and BMDL value. Results The geometric mean of urinary and hair arsenic(98.50 mg/kg Cr, 7.42 mg/kg) μg/L) in the exposed group were significantly higher than urinary and hair arsenic (22.98 mg/kg Cr, 1.28 mg/kg) and each biomarker in the control group(4.63 μmol/L, 13.76 U/L,36.45 U/L,54.62 μg/L,74.45 μg/L,54.81 μg/L, P<0.01). Significant dose-effect relationship existed between urinary and hair arsenic contents and each biomarker. BMD and BMDL value of urinary arsenic was 49.53-101.96 mg/kg Cr and 39.02-70.15 mg/kg Cr, respectively. Those of hair arsenic were 3.04-5.02 mg/kg and 2.36-3.25 mg/kg, respectively. According to BMD and BMDL value of urinary and hair arsenic, the sensitivity of biomarkers decreased in the order of GSTs, TBA and Conclusions According to the lowest BMD and BMDL of urinary and hair arsenic, averaged reference value of urinary and hair arsenic in the local normal population, we suggest urinary 35.0 mg/kg Cr and hair 2.5 mg/kg as their biological exposure limits for those with liver dysfunction induced by arsenic-coal burning. GSTs, TBA, γ-GT and HA, Ⅳ. C, PC-Ⅲ can respectively reflect liver cell damage and liver fibrosis caused by arsenic-coal burning in different degrees, among which, GSTs and HA are the most sensitive biomarkers respectively for liver cell damage and liver fibrosis.

OBJECTIVE: To verify the clinical effect of acupuncture at "experienced ten acupoints" for functional dyspepsia (FD) of liver stagnation and spleen deficiency. METHODS: A total of 96 patients with FD of liver stagnation and spleen deficiency were randomly divided into an observation group and a control group, 48 cases in each group. In the observation group, "experienced ten acupoint" prescription was applied，acupoints included Shangwan (CV 13), Zhongwan (CV 12), Xiawan (CV 10), Qihai (CV 6), Tianshu (ST 25), Zusanli (ST 36) and Neiguan (PC 6). In the control group, irrelevant acupoint prescription was applied, acupoints included Feiyang (BL 58), Heding (EX RESULTS: Compared before treatment, the symptom score after treatment was decreased in the observation group ( CONCLUSION Acupuncture at "experienced ten acupoints" can improve symptoms in patients with FD of liver stagnation and spleen deficiency, has the better clinical effect.
Acupuncture Points ; Acupuncture Therapy ; Dyspepsia/therapy* ; Humans ; Liver ; Spleen

OBJECTIVETo establish a primary culture method of rat testis Leydig cell.

METHODSThe primary rat Leydig cells were treated with or without 4 U/ml human chorionic gonadotropin (hCG), and testosterone in culture medium was detected by radioimmunoassay. The morphology and biological characteristics of Leydig cell were observed.

RESULTSThe culture cells were highly homogeneous, proliferative and had a high differentiation rate. The high purified Leydig cells were verified by their dynamic morphological changes and 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD) histochemical staining. The testosterone secretion induced by hCG significantly increased (P < 0.05) 24 hours after inoculation than that induced without hCG in the control.

CONCLUSIONIt suggests that the Leydig cell cultured in vitro may secrete high concentration of testosterone, and this study laid the basis of androgen replacement therapy for partial androgen deficiency in aging male.

3-Hydroxysteroid Dehydrogenases ; Animals ; Cell Culture Techniques ; Cells, Cultured ; Chorionic Gonadotropin ; pharmacology ; Humans ; Leydig Cells ; cytology ; drug effects ; secretion ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; cytology ; drug effects ; Testosterone ; metabolism

OBJECTIVETo study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.

METHODSTo construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.

RESULTSAfter transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.

CONCLUSIONGrowth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Blotting, Western ; Brain Neoplasms ; pathology ; therapy ; Cell Line, Tumor ; Cell Proliferation ; DNA, Antisense ; genetics ; metabolism ; Flow Cytometry ; Genetic Therapy ; methods ; Glioma ; pathology ; therapy ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Telomerase ; genetics ; metabolism ; Transfection ; Tumor Burden ; Xenograft Model Antitumor Assays

OBJECTIVETo explore the effects of Huoxiang Zhengqi liquid (HXZQ) on enteric mucosal immune responses in mice with Bacillus dysenteriae and Salmonella typhimurium induced diarrhea (BSD).

METHODMice were randomly divided into four groups with 10 mice in each group: control group (control), BSD group, Huoxiang Zhengqi liquid treated BSD groups at high dosage and low dosage (HXZQ high, HXZQ low). HXZQ was administrated from the day of diarrhea induction at dosage of 5.21 g kg(-1) and 0.52 g kg (-1) respectively. Peyer's patch and periphery lymphocytes were prepared for flow cytometry, and level of TNF-alpha in periphery and enteric tissue homogenate were determined with ELISA. Student's t-test was used for statistics.

RESULTMice in BSD group started showing continuous diarrhea at the day of induction till the fourth day when the mice were sacrificed. Diarrhea in the mice of HXZQ high and low groups lasted for 36 and 54 h respectively. There were more CD4+ and CD8+ cells in periphery, less CD4+ cells in peyer's patch in BSD mice comparing to normal mice. In peyer's patch, there were more CD8+ cells in mice in HXZQ high and low groups and more CD4+ in mice in HXZQ high group. Higher level TNF-alpha in periphery and intestinal tissue homogenate in BSD group were observed. Mice in HXZQ high group showed the decreased level TNF-alpha in periphery and enteric tissue homogenate.

CONCLUSIONThe immune regulation on peyer's patch CD4+ and CD8+ cells and suppression on TNF-alpha level in enteric homogenate might partially explain the effect of HXZQ on improvement of BSD.

Animals ; CD4-CD8 Ratio ; Colon ; immunology ; metabolism ; pathology ; Diarrhea ; immunology ; metabolism ; microbiology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Dysentery, Bacillary ; immunology ; metabolism ; microbiology ; Immunity, Mucosal ; drug effects ; Intestinal Mucosa ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peyer's Patches ; drug effects ; immunology ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Salmonella Infections ; immunology ; metabolism ; microbiology ; Salmonella typhimurium ; immunology ; Shigella dysenteriae ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; blood ; metabolism