Objective To investigate the anticancer effect of oxymatrine on cervical cancer cell line (HeLa). Methods MTT assay was used to detect the anti-proliferative effect of oxymatrine.Transwell chamber was used to detect the anti-metastatic effect of oxymatrine.Real-time PCR was used to detect the mRNA levels of MMP-2 and MMP-9.Western blot was used to detect the protein levels of MMP-2,MMP-9,AKT,p-AKT and GADPH. Results We found that application of oxymatrine significantly inhibited the growth of HeLa cells at the concentration above 0.8 mg/mL.We also found that oxymatrine (0.1,0.2 and 0.4 mg/mL)inhibited the invasion of HeLa cells under cytotoxic dose,which was (77.07±20.43)%,(53.95±18.17)% and (20.35±11.20)% of cells that migrated through the matrigel when compared with those of non-oxymatrine treatment group (P<0 .0 5 ). Further research found that oxymatrine (0.1,0.2 and 0.4 mg/mL)could reduce the expression of MMP-2 at the mRNA level,i.e.(82.76±8.71)%,(39.51±12.79)% and (21.53±5.38)% of the expression level when compared with that of non-oxymatrine treatment group (P<0 .0 5 ).The protein expression level of MMP-2 in 0 .4 mg/mL group was (64.69 ±16.52)% of non-oxymatrine treatment group (P<0.05).The phosphorylation level of AKT in 0.4 mg/mL group was (41.27±7.13)% of non-oxymatrine treatment group (P<0.05).Conclusion Oxymatrine can inhibit the invasion of HeLa cells by reducing the expression of MMP-2 via inhibiting the activity of AKT signal pathway.All together,our findings bring new insights into the mechanism of the anticancer effects induced by oxymatrine treatment.

Objectives To observe the changes of magnetic resonance imaging(MRI) during the course of treatment in hepatolen-ticular degeneration (HLD) and to evaluate its related factors.Methods Sixty - three patients with HLD received routine SE sequence MRI of brain and liver.Twenty - two patients with abnormal MRI in 42 patients and each of them had 2-4 times of serial MRI ex-aminations at interval of 1.0- 1.5 year.Results Ten patients had lesion in both brain and liver in 22 patients. Nine patients only in brain, 3 patients in liver.The major lesions disappear or decrease after rational treatments. There were only slight or no changes of MRI abnormalities in patients receiving treatments or age of patients was over 15 year old. Abnormal MRI findings shown again after stopped treatments over 2 months. In such cases, the clinical and MRI improvements were slow.The rate of clinical and MRI improvement in patients with adequate treatment were markedly higher than patients with inadequate treatment.Conclusions Liver and brain lesion will show in MRI disappear after adequate treatment;There are only slight or no change or showing again in MRI abnormalities in pa-tients with inadequate treatments compared with poor clinical improvement. MRI is beneficial to understanding therapeutic effect and prognosis of HLD.

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

AIM: To investigate the potassium channel gene expression of myocardial sleeves of pulmonary vein and effects of amiodarone on rabbits with rapid atrial pacing.METHODS: Rabbits were divided into three groups(n=10),(1) the control group with sham operation and placebo;(2) the right atrial pacing(RAP) group at 600 beats/min with the placebo;(3) the amiodarone group treated for seven days with oral amiodarone at 100 mg ? kg-1 ? d-1.Based on RAP simultaneously,the messenger ribonucleic acid(mRNA) of specimen was measured by reverse transcription-polymerase chain reaction.RESULTS: Compared with the control group,Kv4.3(transient outward K+ current,Ito1) mRNA expression in RAP group was reduced by 51%(P

Objective To evaluate the efficacy and safety of colistimethate sodium (CMS) for the treatment of critical patients infected by pan-drug resistantAcinetobacter baumannii (PDR-AB) or pan-drug resistant Pseudomonas aeruginosa (PDR-PA).Methods 321 isolates of PDR-AB and 204 isolates of PDR-PA from critical patients admitted to 35 intensive care units (ICUs) of grade two or above were collected from the Anhui Antimicrobial Resistance Investigation Net (AHARIN) program from September 2012 to September 2015, while the minimal inhibitory concentrations (MIC) of colistin were determined by the E-test. A series of Monte Carlo simulations was performed for CMS regimens (1 MU q8h, 2 MU q8h, and 3 MU q8h, and MU meant a million of unit), and the probability of achieving a 24-hour area under the drug concentration time curve (AUC24)/MIC ratio > 60 and risk of nephrotoxicity for each dosing regimen was calculated. Each simulation was run over three CLCr ranges: < 60, ≥ 60-90, ≥ 90-120 mL/min. The probability of target attainment (PTA)for the AUC24/MIC ratio was calculated using the partial MIC value, while the cumulative fraction of response (CFR) was determined by integrating each PTA with the MIC distributions, the value greater than or equal to 90% or more than 80% was set as the optimal dosing regimen or suboptimal dosing regimen respectively. The probability of average 24-hour serum concentrations up to 4 mg/L for three dosage regimens was used to predict the risks of nephrotoxicity.Results All 321 isolates of PDR-AB and 204 isolates of PDR-PA were susceptible to colistin, the MIC50/90 against PDR-AB were 0.5mg/L and 1.0 mg/L, and those against PDR-PA were 0.5 mg/L and 1.5 mg/L, respectively. When recommended dose (1 MU q8h) was used for patients with CLCr of < 60 mL/min, high CFR value (89.78% for PDR-AB, 81.06% for PDR-PA) were obtained, but with a high risks of nephrotoxicity (> 32.51%). Moreover, low value of PTA (< 66.56%) was yielded for isolates with MIC of ≥ 1 mg/L. Recommended dose also yielded a low CFR value (56.97%-69.31% for PDR-AB, 44.76%-56.94% for PDR-PA) in patients with CLCr of ≥ 60-120 mL/min. When dose was increased to 2 MU q8h, CFR (77.45%-92.87%) and the risks of nephrotoxicity (< 0.15%) was optimal for patients with CLCr ≥ 60-120 mL/min, but low value of PTA (< 75.36%) was also yielded for isolates with MIC of ≥ 1 mg/L. The most aggressive dose of 3 MU q8h provided high CFR (> 89.24%) even in patients with CLCr ≥ 90-120 mL/min, and PTA was < 76.20% only for isolates with MIC of ≥ 1.5 mg/L, but this dosing scheme was associated with unacceptable risks of nephrotoxicity (> 33.68%).Conclusion Measurement of MIC, individualized CMS therapy and therapeutic drug-level monitoring should be considered to achieve the optimal drug exposure and ensure the safety of CMS.

Objective To establish the methods for quantitating the circulating tumor DNA with PicoGreen fluorescent nucleic acid stain and investigate the role of the quantitative analysis in diagnosis of breast cancer. Methods Circulating tumor DNA was isolated from serum with QIAmp blood kit and was quantitated by spectrofluorometry with PicoGreen fluorescent stain. Receiver operator characteristic ( ROC) curve and area under the curve were used to estimate the role of DNA quantification in diagnosis of breast cancer. Results The efficiency of QIAamp blood kit isolating DNA from serum was 37. 8%-46. 2%, average 43. 4%. Circulating tumor DNA concentration as low as 1 ng could be detected by PicoGreen spectrofluorometry. and the detected range was 1-500 ng/0. 2 ml. The median concentration of serum DNA in breast cancer group was (169. 70+ 124. 10) μg/L, and that of healthy control and breast benign group were (54. 30±36. 84) μg/L and (51. 70±29. 04) μg/L, respectively (P<0. 01). The area under the ROC curve was 0. 899 (95% CI: 0.848-0.951), and the sensitivity was 78. 2%, the specificity was 90% by using the cutoff value of 96. 0 ng/ml. Conclusions The concentration of circulating tumor DNA can be efficiently quantitated by PicoGreen spectrofluorometry, which indicates the potential of clinical applicability in breast cancer diagnosis.

The safflower floret is a traditional Chinese medicine used to promote blood circulation and remove obstruction in the channels. The spines on its bracts are considered a handicap when manual harvest is involved. In this study, cDNA-SRAP was used to systematically investigate which genes are associated with the spines. Sixty pairs of possible primer combinations were used on two cDNA pools representing spininess and spinelessness. Six transcript-derived fragments were identified, of which two with low recombination were sequenced successfully and named as GPY-1 and GPY-2. By using the RACE method, the full-length cDNA of GPY-2 is cloned and named as CTL-spn. The full-length cDNA of CTL-spn was 1 679 bp long with a 1 524 bp ORF encoding a 508 aminoacid protein. The deduced amino acid sequence of the CTL-spn gene shared a high homology (97%) with other known ATP synthase CF1 alpha subunits. Semiquantitative RT-PCR analysis revealed that the mRNA of GPY-1 and GPY-2 accumulated in only spiny lines. Considering the important role of ATP synthase CF1 alpha subunit in plants, it may directly take part in the formation process of spininess and enhancing resistance reaction of spiny safflower. Also, our results provide the important insights for breeding spineless cultivars of safflower.
Adenosine Triphosphate ; Amino Acid Sequence ; Carthamus tinctorius ; enzymology ; genetics ; Chloroplast Proton-Translocating ATPases ; genetics ; DNA Primers ; DNA, Complementary ; Plant Proteins ; genetics

Effect of p-nitrophenol shock on microbial populations and sludge activity in UASB reactor was studied by DGGE-PCR of 16S rDNA fragments and detection of COD removing and biogas yield.The results showed that p-nitrophenol seriously inhibited the sludge activity,resulting in the drop of biogas and COD removing rate.The 40mg/L p-nitrophenol had more inhibition than 20mg/L p-nitrophenol.It would take 27 and 16 days respectively for reactor to recover after 40mg/L and 20mg/L p-nitrophenol shock.The diversity of eubacteria and methanogens were also effected by the p-nitrophenol shock.The variation of eubacteria was more than that of methanogens after p-nitrophenol shock.The drop of biogas was mainly related to the variation of Methanosaeta sp.and Methanomicrobia sp.after p-nitrophenol shock.Among the eubacteria the population of Chloroflexi sp.、Bacteroide sp.and Anaerovibrio sp.decreased greatly after p-nitrophenol shock.And more,the Rheinheimera sp disappeared after 40mg/L p-NP treatment.But the Flavobacteria sp.appeared after p-nitrophenol shock,which was probably related to the degradation of p-NP.

ObjectiveTo investigate the changes of the morphology, growth and cycle of CHO(dhfr-) cells after space flight.MethodsCHO(dhfr-) cells were carried in the No.18 recoverable satellite and monocloned harvesting cells before multiplying. 4 cell lines were selected randomly,and the growth characteristics of the most slowly growing one at the fifth passage was observed by methods of MTT and FCM as well as the cells' shape.Results159 cell strains were obtained after monocloning and multiplying. The cells' morphology changes, growth speed decrease and the number of G1 phase increased markedly.ConclusionSpace flight induced morphological changes of cells and it is impossible to screen out finer bioengineering cells.

AIM: To study caspase-9 expression on rat retina in the process of chronic elevation of IOP and the changes with the application of amino guanidine (AG), thus to investigate potential protective function of AG to rat retina with chronic elevation of IOP.METHODS: Immunohistochemistry, RT-PCR and Western blot were used to observe retinal morphology and expression of caspase-9 at different time points of rat with chronic IOP elevation, both affected or not affected by the application of AG.RESULTS: Compared with control group, as time passed retina of experimental group gradually had detectable morphological changes. On 21st day of chronic IOP elevation, retinas became thinner and the quantity of retinal ganglion cells (RGCs) decreased; caspase-9 expression increased, consistent with the morphological changes. The group using AG presented relatively smaller morphology changes and less expression of caspase-9.CONCLUSION: Apoptosis-related gene caspase-9 played a part in the process of chronic IOP elevation; AG protects retina by down-regulating expression of caspase-9.