Objective To establish a HPLC method for the determination of ferulic acid in Xuefu Zhuyu Decoction. Methods A Kromasil C18(250 mm ×4. 6 mm, 5 μm) column was adopted. The mobile phase consisted of acetonitrile-0. 085 % phos-phoric acid(17 : 83)with the flow rate of 1.0 mL/min. The column temperature was at 35 ℃ and detection wavelength was set at 316 nm. Results A good linearity of ferulic acid was in the range of 0. 0252μg to 0. 504μg and r=0. 999 9. The average recovery was 100. 15 % and RSD=1.69 %. Conclusion The method is simple and rapid, and it is suitable for the determi-nation of ferulic acid in Xuefu Zhuyu Decoction.

[Objective] Antifungal action of fat-soluble extracts from Bulbus Allii Cepae (BAC) by different extraction technology on 16 strains of dermatophytes was observed to supply evidence for the optimization of extraction technology for BAC. [Methods] Four methods were used for the extraction of fat-soluble extract from BAC and the antifungal action of the extract was tested by medium dilution method. [Results] Fat-soluble extract freshly extracted from BAC by ether had a strong in-vitro antifungal action and its minimum inhibitory concentration (MIC) was not over 6.25 mL?L-1 against 10 strains of fungi including Shanghai Epidermophytom floccosum and its MIC against other strains ranged 12.5 -50.0 mL?L-1. The minimum fungicidal concentration of fat-soluble extract freshly extracted from BAC by ether was as the same as MIC. Fat-soluble extract freshly extracted by ether had no antifungal action after water removal by heating. [Conclusion] The extraction technology of freshly extracted by ether without heating is a better method for BAC and the fat-soluble extract extracted by this method has a better antifungal action.

Aim To investigate the effects of heme oxygenase-1(HO-1) on brain damage due to cerebral ischemia/reperfusion.Methods 120 male mice were divided into four groups randomly and the cerebral ischemia/reperfusion(I/R) model was established through drawing out and reperfusing 40% of the whole blood volume in combination with clamping the carotid arteries for 20 min after anesthesia with 40 g?L-1chloral hydrate(400 mg?kg-1 ip).The mice were intracerebraventricularly(icv) injected with 3(l of150 ?mol?L-1 hemin,150 ?mol?L-1 ZnPPIX,or artificial cerebrospinal fluid(ACSF),respectively 16 h before the model established.Sham operation group only received anesthesia and operation without I/R and icv injection.The hippocampal HO activity and cerebral cortex xanthine oxidase(XO) activity,the levels of malonaldehyde(MDA) and reactive oxygen species(ROS),hippocampal neuron apoptosis,and hippocampal HO-1 expression were determined by spectrophotometer,TUNEL method,and Western blot,respectively.Results The increased hippocampal HO-1 expression and HO activity,decreased XO activity,MDA and ROS levels,and diminished hippocampal apoptosis were observed in the hemin group,compared with ACSF group(P0.05),but inhibited the activity of HO with the elevation of XO activity,MDA and ROS levels,as well as cellular apoptosis(vs ACSF,P

Peroxisome proliferator-activated receptoralpha (PPARalpha) is a ligand-activated transcription factor which plays a pivotal role in the regulations of metabolism. A cDNA encoding ligand binding domain (LBD) of PPARalpha was amplified by RT-PCR from human hepatic tissue and the product was inserted into the downstream of the malE gene in the vector pMAL-p2X,which encodes maltose-binding protein (MBP). The recombinant plasmid containing MBP-PPARalpha gene was transformed into E. coli. TB1, and then the growth conditions of the recombinant strain were studied, which remarkably influenced the final yield of protein expression. With the use of SDS-PAGE and Bio-Rad Quantity One gel image analysis, we found the best expression condition as follows: The induction was started as OD600 reached 0.5 by the adding of IPTG to a final concentration of 0.4 mmol/L, and then the incubation continued 6 hours at 30 degrees C. The maximum yield of fusion protein was 31.34% of the total mass of cytoplasm proteins. Moreover, the soluble form of the target protein is useful for further work on purification and on screening the ligands of PPARalpha.
Carrier Proteins ; biosynthesis ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Ligands ; Maltose-Binding Proteins ; PPAR alpha ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

BACKGROUND:Cervical cancer is one of the most frequent malignancies in women worldwide, and its occurrence and development is closely related to cyclooxygenase-2 (COX-2).OBJECTIVE: To examine the expression of COX-2 alternative splicing variants in human cervical carcinoma tissue and understand its possible implications.DESIGN: Non-randomized controlled experiment.SETTING: Key Laboratory of Biochemistry and Molecular Pharmacology,Department of Obstetrics and Gynecology, First Affiliated Hospital,Chongqing Medical University.PARTICIPANTS: Carcinoma tissue and normal tissue were obtained from 13 cervical carcinoma patients admitted during March 2002 to April 2002in the Department of Obstetrics and Gynecology, First Affiliated Hospital,Chongqing Medical University.METHODS: A pair of specific primers were designed for reverse transcription-polymerase chain reaction (RT-PCR) to obtain the mRNA of COX-2 in human cervical carcinoma tissues. The resultant band on electrophoresis was cloned, sequenced and analyzed.MAIN OUTCOME MEASURES: ① Agarose gel electrophoresis result of the PCR product of carcinoma and normal tissues; ② Sequencing result of the electrophoresis band from carcinoma and normal tissues.RESULTS: No COX-2 band (252 bp) was found in electrophoresis for normal tissues, while 2 bands appeared for cervical carcinoma tissues, including a new electrophoresis band of 534bp besides the COX-2 band. Cloning and sequencing revealed that this new band contained not only exons 7and 8 of COX-2 gene but also a reserved intron of 282 bp intron between exons 7 and 8. Analysis of the predicted amino acid sequence indicated that an in-frame stop codon occurred in the 48-50 bp of the intron retained in the mRNA.CONCLUSION: The presence of COX-2 alternative splicing mRNA variant (Genbank accession number:BU493602)is confirmed in human cervical carcinoma tissue, which codes for a protein possibly smaller than COX-2.

Objective To discuss of the therapeutic effects of treating three TCM types of Chronic superficial gastritis (CSG)disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach with chinese medicine, and to discuss its impact on the changes of three gastrointestinal hormones-- motilin (MTL), gastrin (GAS), and somatostatin (SS). Methods The data of 150 CSG patients in Shenzhen Bao’an Hospital from Feb, 2011 to Mar, 2013 were selected. These patients belonged to three kinds of syndrome, which was disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach, each syndrome had 50 cases. Different Chinese medicines were applied to different syndrome of patients. After one month treatment, the therapeutic effects and the changes of MTL, GAS, SS in these patients were detected by radioimmunoassay. Results After treatment, the total effective rate of the damp heat in the spleen and the stomach syndrome was 94%, the weakness of the spleen and the stomach type was 88%, disharmony of liver and stomach type was 90%, the difference was not statistically significant among the three types(χ2=3.84, P>0.05);after treatment, the serum MTL, SS level of disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach type were [damp heat in the spleen and the stomach type were (240.25±7.29)pg/ml, (68.23± 5.97)pg/ml;the weakness of the spleen and the stomach type were (260.16±8.84)pg/ml, (75.35±4.65)pg/ml;disharmony of liver and stomach type were(258.32±15.23)pg/ml, (80.12±7.88)pg/ml] higher than before treatment [damp heat in the spleen and the stomach type were (87.26±7.58)pg/ml, (49.34±4.93)pg/ml;the weakness of the spleen and the stomach type were (67.11±7.81)pg/ml, (57.15±5.11)pg/ml; disharmony of liver and stomach type were (89.26±7.60)pg/ml, (40.63±7.26)pg/ml], and the levels of serum GAS [(156.38±17.36)pg/ml, (146.15±15.56)pg/ml, (140.26±13.37)pg/ml] was lower than before treatment [(274.59 ± 17.82)pg/ml, (320.83 ± 15.02)pg/ml, (216.15 ± 13.77)pg/ml], the difference was statistically significant (P<0.05 or 0.01). Conclusions The changes of three hormones-MTL, GAS, SS can be used as an objective basis for the diagnosis and the judgment of healing efficacy of CSG treated by traditional Chinese medicine.

Objective To evaluate the prevention of glucocorticoid -induced avascular necrosis of the femoral head(ANFH)with Tongluo Shenggu Capsule(TSC).Methods One hundred and twenty adult SD rats were randomized into six groups:normal control group,model group,clofibratum group and three -dose TSC groups(high -,moderate -and low -dose respectively).ANFH rat models were established by injection of glucocorticoid once per week and the med-ication groups were given drugs by ga stric infusion one time a day.The rats were killed in the fourth,eighth and twelfth week respectively.Then the macrosc opic and microscopic feature of the f emoral head were observed,and ink -p erfused specimens of the femoral head were detected and analyzed by auto -photograph software of Metamoph to evaluate the pre-ventive effect of TSC.Results In TSC groups,bone trabecula grew we ll,similar to the normal group;lacu nar bone per-centage was lower than that in the mod el group;fatty degeneration in marrow cavity of the femoral head was redu ced,andthe number of subcartilaginous bloo d vessels and their diameter were increased as compared with those in the model group(P

Objective To compare and analyze the dosimetric discrepancy of combind intracavitary/interstitial brachytherapy using three different kinds of optimization method in locally advanced cervical cancer.Methods Totally 20 cases of locally advanced cervical cancer were selected and divided into three groups according to different optimization method which include manual optimization group (MO) based on graphical optimization,inverse planning simulated annealing (IPSA 1)based on simulated annealing optimization algorithm,IPSA 2 based on IPSA 1 with limitation on maximum dose of target.The dose volume histogram parameters of the targets (V200,V150,V100,D100,D90,HI) and the OARs(D0.1 cm3,D1 cm3 and D2 cm3) were analyzed.Results For CTV,compared with MO,there was no significantly statistical difference in D100between IPSA 1 and IPSA 2(P ＞ 0.05).However,V200,V150,V100 and HI for ISPA1 were better than for ISPA2 (t =-3.422-9.910,P ＜ 0.05).In addition,V100 and D100 in ISPA1 were better than in ISPA2 (t =7.238,5.032,P ＜0.05).For OARs,D0.1 cm3,D1 cm3 and D2 cm3 in rectum,bladder,sigmoid colon of both ISPA 1 and ISPA 2 were dramatically lower than those of MO (t =2.235 5.819,P ＜ 0.05),without significantly statistical difference found between ISPA1 and ISPA2.Conclusions For combined intracavitary/interstitial brachytherapy in locally advanced cervical cancer,all treatment plans based on three different kinds of optimization methods can meet the clinical need.Moreover,inverse optimization can ensure dose coverage over target and reduce maximum dose of rectum,bladder and sigmoid colon.

Objective:To analyze the incidence and epidemiological characteristics of traumatic spinal cord injury in China in 2018.Methods:Multi-stage stratified cluster sampling was used to randomly select hospitals capable of treating patients with spinal cord injury from 3 regions，9 provinces and 27 cities in China to retrospectively investigate eligible patients with traumatic spinal cord injury admitted in 2018. National and regional incidence rates were calculated. The data of cause of injury，injury level，severity of injury，segment and type of fracture，complications，death and other data were collected by medical record questionnaire，and analyzed according to geographical region，age and gender.Results:Medical records of 4，134 patients were included in this study，with a male-to-female ratio of 2.99∶1. The incidence of traumatic spinal cord injury in China in 2018 was 50.484 / 1 million （95% CI 50.122-50.846）. The highest incidence in the Eastern region was 53.791 / 1 million （95% CI 53.217-54.365）. In the whole country，the main causes of injury were high falls （29.58%），as well as in the Western region （40.68%），while the main causes of injury in the Eastern and Central regions were traffic injuries （31.22%，30.10%）. The main injury level was cervical spinal cord in the whole country （64.49%），and the proportion of cervical spinal cord injury in the Central region was the highest （74.68%），and the proportion of lumbosacral spinal cord injury in the Western region was the highest （32.30%）. The highest proportion of degree of injury was incomplete quadriplegia （55.20%），and the distribution pattern was the same in each region. A total of 65.87% of the patients were complicated with fracture or dislocation，77.95% in the Western region and only 54.77% in the Central region. In the whole country，the head was the main combined injury （37.87%），as well as in the Eastern and Central regions，while the proportion of chest combined injury in the Western region was the highest （38.57%）. A total of 32.90% of the patients were complicated with respiratory complications. There were 23 patients （0.56%） died in hospital，of which 17（73.91%） died of respiratory dysfunction. Conclusions:The Eastern region of China has a high incidence of traumatic spinal cord injury. Other epidemiological features include high fall as the main cause of injury cervical spinal cord injury as the main injury level，incomplete quadriplegia as the main degree of injury，head as the main combined injury，and respiratory complications as the main complication.