Objective: To explore the expression of KIF20A (kinesin family member 20A) in colorectal cancer (CRC) tissues and adjacent normal tissues, and to analyze the relationship between KIF20A expression level and clinicopathological factors in CRC patients. Meth-ods: Data from The Cancer Genome Atlas (TCGA) database were used to analyze KIF20A mRNA expression in CRC tissues and adjacent normal tissues. A total of 105 paraffin samples were obtained from CRC patients who had undergone surgery at Huai He Hospital of Henan University, from January 2011 to December 2012. Immunohistochemical staining (IHC) was performed to examine KIF20A pro-tein expression in tumor samples for which complete clinical and pathological data were available. Statistical analyses were applied to analyze the association between KIF20A expression and the clinical data, as well as with survival outcomes. Results: Bioinformatics analysis showed that the mRNA expression level of KIF20A was upregulated in CRC tissues and normal tissues (P<0.001). IHC revealed significantly higher expression of KIF20A in CRC tissues from 67 patients (64%) and lower or undetectable expression in 38 patients (36%). The difference was statistically significant (P<0.05). Overexpression of KIF20A in CRC tissues was significantly associated with depth of invasion, lymphatic node metastasis, distant metastasis, and TNM stage (all P<0.05). Kaplan-Meier survival analysis showed that patients with high levels of KIF20A expression had poor prognosis compared to patients with low levels of KIF20A expression. Cox proportional hazard regression analysis revealed that KIF20A was an independent prognostic factor in patients with CRC. Conclusions:KIF20A is upregulated in CRC tissues and could serve as a novel prognostic biomarker for CRC patients.

Chinese medicine processing is the main feature that distinguishes traditional Chinese medicine from natural medicine and plant medicine, and is the main feature in clinical medication of traditional Chinese medicine. The research of Chinese medicine processing technology is an important link to realize standardization and standardization of Chinese herbal pieces, with urgent need to attract high attention. At present, there are still many problems in the research of processing technology of Chinese herbal pieces, mainly including inconsistent processing technology, large differences in process technology parameters, and unstable production technology of Chinese herbal pieces, resulting in uncontrollable quality of Chinese herbal pieces and affecting the clinical efficacy of Chinese medicine. This paper focused on the establishment of a unified standard processing technology, and put forward the countermeasures for the processing technology of Chinese medicine based on a comprehensive analysis of the current situations of the processing technology of Chinese herbal pieces, with significance for guiding the establishment of a standardized processing technology of Chinese medicine.

Objective To analyze the clinical and electrophysiological features in a family with spinal muscular atrophy (SMA), and assess the probable causative gene mutations for the family. Methods To identify the nosogenesis of the proband with weakness and atrophy in the double lower proximal limbs, clinical data of his 12 family members were collected, and the proband and his mother were selected for clinical examinations, including laboratory tests, electromyogram (EMG), F-wave, H-reflex, X-ray of the spine and double lower limbs, brain and spinal cord magnetic resonance imaging, etc. Moreover, human whole exome sequencing was performed on blood sample from the proband, then its deleterious effects were assessed according to the Standards and guidelines for the interpretation of sequence variants, a joint consensus recommendation of the American College of Medical Genomics (ACMG) and the Association for Molecular Pathology (AMP). Subsequently, the strong pathogenic mutation was validated by Sanger sequencing. Results Familial investigation showed seven of 12 family members presented with weakness in the double lower proximal limbs. Among them, three had the main manifestation of atrophy in the double lower proximal limbs, one had high arched foot as the main presentation, and the others had weakness in the double lower proximal limbs. EMG studies showed the abnormal results in the anterior horn of the spinal cord. The strong pathogenic mutation in DYNC1H1 gene (exon8, c.2327C＞T, p.P776L) was identified from the proband according to ACMG and AMP guidelines. Sanger sequencing revealed six patients had this variant and it was passed mainly from his maternal grandmother. Conclusions A pathogenic mutation of the DYNC1H1 p.P776L in six Chinese pedigrees which cosegregated with SMA was identified. There existed individual differences in clinical presentations. This finding may have important implications for the study of SMA in Chinese patients.

The present study was designed to optimize the processing of Fructus Arctii by response surface methodology (RSM). Based on single factor studies, a three-variable, three-level Box-Behnken design (BBD) was used to monitor the effects of independent variables, including processing temperature and time, on the dependent variables. Response surfaces and contour plots of the contents of total lignans, chlorogenic acid, arctiin, and arctigenin were obtained through ultraviolet and visible (UV-Vis) monitoring and high performance liquid chromatography (HPLC). Fructus Arctii should be processed under heating in a pot at 311 °C, medicine at 119 °C for 123s with flipping frequently. The experimental values under the optimized processing technology were consistent with the predicted values. In conclusion, RSM is an effective method to optimize the processing of traditional Chinese medicine (TCM).
Arctium ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Furans ; analysis ; Glucosides ; analysis ; Hot Temperature ; Lignans ; analysis ; Surface Properties ; Technology, Pharmaceutical ; methods

AIM: To develop a high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) and ultraviolet (UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang (DFT). METHOD: The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL·min(-1) and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV. RESULTS: All calibration curves showed good linear regression (r(2) ≥ 0.9990). The limits of detection and limits of quantification were 0.021-0.155 -g·mL(-1) and 0.076-0.520 -g·mL(-1), respectively. Overall precision RSD (intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%-101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.
Aconitum ; chemistry ; Asarum ; chemistry ; Calibration ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Limit of Detection ; Quality Control ; Reproducibility of Results ; Rheum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spectrophotometry, Ultraviolet ; methods

Objective To establish and evaluate unstable atherosclerotic plaque model in abdominal aorta induced by cold stress. Methods Sixty male New Zealand white rabbits were randomly divided into three groups:cold stress group fed with high fat diet and followed by balloon induced arterial wall injury of abdominal aorta at week 2 and exposed to cold (4℃) for 1 h per day except for the first postoperative week,balloon-injury group treated by high fat diet plus balloon-injury, control group fed a normal chow without any treatment. Pathological changes of atherosclerotic plaques among these groups were evaluated at 20 weeks. Meanwhile, serum concentrations of blood lipid,oxidized low density protein(ox-LDL),hypersensitive C-reaction protein (hs-CRP)and interleukin (IL)-8 were determined. Results There was no difference in blood lipid level between cold-stress and balloon-injury groups.Serum concentrations of ox-LDL[(56.1 +14.3)mg/L vs.(42.9± 13.8)mg/L],hs-CRP[(149.1+78.3)mg/L vs. (94.5±57.3)mg/L],IL-8 [(97.6±17.9)μg/L vs.(57.5±18.3)μg/L]and macrophage infiltration[(30.9±5.6)％ vs,(18.7±4.8) ％] were significantly higher in cold stress group than in balloon-injury group (t =2.78,6.91,14.94,6.88,all P＜0.05). Higher angiogenesis rate of atherosclerotic plaque in cold-stress group (23/31,74.1％) was observed in comparison with group balloon-injury(5/25,20,0％)(x2=16.26,P＜0.05). Conclusions Establishment of rabbit unstable atherosclerotic plaque model induced by cold stress in synergy with high fat diet and balloon-injury is feasible, which is superior to conventional method through high fat diet plus balloon-injury surgery.

To explore new microbial resources in deep subsurface oil reservoirs, strain DL-7 was isolated with Hungate technology from oil reservoir water sampled from Dagang oilfield, China. Physiological and biochemical examinations showed that H2/CO2 is the unique substrate of the strain, which cannot metabolize formate, methanol, trimethylamine, acetate and other secondary alcohols. The optimum growth conditions were further identified to be 60 degrees C, pH 7.0-7.5 and 0.25% NaCl. Moreover, the strain cannot grow without yeast extract. Analysis of its 16S rRNA sequence indicated that a similarity of 99.7% presents between the strain and the model species M. marburgensis DSM2133T (X15364).
Hot Temperature ; Methanobacteriaceae ; classification ; genetics ; isolation & purification ; metabolism ; Methanol ; metabolism ; Methanomicrobiaceae ; genetics ; isolation & purification ; Petroleum ; microbiology ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Water Microbiology

Objective To investigate the effects of low frequency pulsed magnetic fields (LF-PMFs) on proliferation, the cell cycle, apoptosis, migration, cytoskeleton formation and nitric oxide (NO) secretion in cardiac microvascular endothelial cells (CMECs). Methods LF-PMFs with rectangular and triangular waveforms at 15 Hz were tested. The CMECs were randomly divided into a control group, 0.6 mT group, 1.2 mT group, 1.8 mT group and 2.4 mT group. Except for the control group, the CMECs were exposed to LF-PMFs 4 h/day for 5 days. Results After 5 days of intervention with rectangular wave LF-PMFs the proliferation of CMECs accelerated and NO secretion was enhanced significantly. The percentages of cells at ( S+G2 ) phase increased significantly, whereas apoptosis rates were significantly lower than in the control group. The migration of CMECs was facilitated, stress fibers increased and cytoskeleton components were reorganized. After 5 days of intervention with triangular wave LF-PMFs proliferation of CMECs accelerated and NO secretion was significantly enhanced. The percentages of cells at(S+G2)phase and migration increased, while apoptosis was inhibited. Cytoskeleton components were reorganized after exposure to 0.6 mT and 2.4 mT triangular waveform LF-PMFs. No significant change was detected with 1.2 mT and 1.8 mT triangular waveform LF-PMFs with regard to these variables. Conclusions The effects of LF-PMFs on proliferation, the cell cycle, apoptosis, migration, cytoskeleton formation and NO secretion function of CMECs were infiuenced by the waveform in addition to it's intensity and frequency.

Objective To determine which parameters of reverse remodeling of left ventricular could become the effective indexes in evaluating the short-term therapeutic effect of cardiac resynchronization therapy(CRT)in heart failure(HF)patients.Methods CRT was performed in 26 HF patients with dysfunctions of wall motion.Serial echocardiography was practiced at baseline,one and three months after CRT.The parameters including left atrial end-diastolic diameter(LADD),left ventrieular end-diastolic diameter(LVDD),end-diastolic volume (LVEDV),end-systolic volume(LVESV),ejection fraction(LVEF)were measured and compared before and after CRT therapy.Results At one and three months after CRT,respectively,CRT was associated with reduced LADD,LVDD,LVEDV,LVESV and improved LVEF and filling time compared with those at baseline.Moreover,all these parameters had better correlations with the activities of these patients.Conclusions LADD,LVDD,LVEDV,LVESV and LVEF could become the effective indexes in evaluating the short-term effect of CRT in HF patients.The reduced diameters and volumes of left atria and left ventricle are more sensitive parameters than the improved LVEF.

BACKGROUND: Angiotensin Ⅱ has been found to induce atrial electrical remodeling, which can be blocked or inhibited by allicin.OBJECTIVE: To study the effects of allicin on angiotensin Ⅱ-induced calcium channel current and intracellular free calcium concentration in human atrial myocytes.DESIGN: A randomized controlled study based on human atrial myocytes freshly isolated.SETTING: Cardiology department of a military medical university of Chinese PLA.METHODS: This study was carried out from June 2003 to June 2004 in the Laboratory of Cardiology Department, Xijing Hospital, the Fourth Military Medical University of Chinese PLA. Ten patients with congenital heart disease who underwent extracorporeal circulation surgery were included in the study. Among them, there were 6 males and 4 females with the average age of 15 ± 6 years. Tissue samples were taken from their right auricle and sent to the lab, where the atrial myocytes were freshly isolated. There were four co-administration of angiotensin Ⅱ (0. 1 μmol/L)and allicin(50 μmol/L).The conventional whole-cell configuration of the patch-clamp technique was used to detect membrane electric current of Ca2 + in L type. Confocal microscope was used with Fluo-3/AM as calcium indicator to detect changes of intracellular free calcium concentration immediately and 15 minutes after drug intervention, respectively.MAIN OUTCOME MEASURES: The peak density of electric current of Ca2 + in L type and alteration of fluoresence intensity of intracellular free calcium concentration.electric current of Ca2 + in L type in human atrial myocytes was significantly increased by angiotensin Ⅱ of 0. 1 μmol/L[( - 12. 77 ± 1. 61) vs ( -5.78affect electric current of Ca2+ in L type in human atrial myocytes group, the peak density of electric current of Ca2 + in L type was significantly lower than that in angiotensin Ⅱ group[ ( - 8.75 ± 0.97) pA/pF, P ＜ 0. 05 ].in angiotensin Ⅱ group was significantly higher than that in control and allicin groups[(2 610.1±112.6, (299.2±27.3)%; 653.9±42.5, 0;simultaneously with angiotensin Ⅱ, the alteration of intracellular fluoresence intensity was much lower than that in angiotensin Ⅱ group[ ( 1284.9 ± 85.2,(96.5±8.4)%;P ＜0.05].CONCLUSION: Allicin antagonizes angiotensin Ⅱ-induced increase in the peak density of electric current of Ca2+ in L type and intracellular calcium overload, which may relieve atrial electrical remodeling.