Two sets of primers were designed according to the sequences of xynA and xynB from Aspergillus sulphureus, and the DNA fragments composed of 574 bp and 594 bp were amplified by polymerase chain reaction (PCR), respectively. The two PCR products respectively digested with EcoRⅠ/BamHⅠ and BglⅡ/HindⅢ were ligated into multiple cloning sites of pET-28a(+). The resulting plasmid is pET-xynAB, in which xylanase A and B are ligated by a 7-amino acid peptide (GlyGlyGlySerGlyGlyGly). E. coli BL21 transformed with pET-xynAB was induced by IPTG, and a special protein band about 50 ku was detected by SDS-PAGE. The protein purified with Ni-NTA column was denatured by 8 mol/L urea and dialyzed for refolding. The recombinant xylanase showed optimal activity at 50℃ and pH 4.4. The enzyme retained above 75% of its activity at the range of pH 2.4～5.4. The xylanase displayed about 50% retained acitivity after incubating at 80℃ for 30 min. Various metal ions have different effects on activity of the recombinant xylanase.

OBJECTIVE:To establish a method for the content determination of muscone in Aiweixin oral liquid. METHODS:After extracting sample by butanol,GC-MS was performed on the column of BD-17 MS with the volume temperature of 270 ℃;the carrier gas was helium with the sample size of 1 μl at the flow rate of 10 ml/min;the split ratio was 20∶1. EI was ion source with the electron energy of 70 eV,ion source temperature of 230 ℃ and quadrupole temperature of 150 ℃;the tuning mode was automatic tuning by quality full scanning with the threshold value of 30 in the range of 30-600 aum. RESULTS:The linear range of muscone was 12.216-61.08 ng(r=0.999 6);RSD of precision test was lower than 2% and the RSDs of reproducibility and stability tests were lower than 3%;the average recovery was 100.24%(RSD=1.58%,n=6). CONCLUSIONS:The method is sensitive, rapid and simple,and can be used for the content determination of muscone in Aiweixin oral lipid.

Objective To study the quality control methods for dragon's blood spraying film agent. Methods The pH value and viscosity of dragon's blood spraying film agent were detected. Drug dispersed homogeneous degree and particle sizes were determined with Nano Particle Size Analyzer and microscope. Content of Loureirin B was measured by Ultra Performance Liquid-Chromtography (UPLC). UPLC was performed on Waters C18 column (2. 1 mm×100 mm,1. 7 μm), the wavelength was 280 nm, the column temperature was 40 ℃ , and the mobile phase was 0. 1% formic acid aqueous solution and acetonitrile, and the flow rate was 0. 8 mL·min-1 . Results The pH value and viscosity of dragon's blood spraying film agent were stable, drug dispersion was homogeneous, and particle size of the drug was tiny. The concentration of Loureirin B had a good linear relationship in the range of 15. 51-77. 54 μg. Conclusion This method can be accurately controlled, has good stability and repeatability, and can fully control quality of dragon's blood spraying film agent.

Objective To investigate the therapeutical effect of Resina draconis. on endometriosis rat model. Methods Totally, 60 adult rats were selected and divided into blank control group( n=10) and blood stasis endometriosis model group( n=50 ) . The blood stasis endometriosis model group was randomly divided into 5 groups after modeling: the Resina draconis. high-, medium-, and low-dose group, Guizhi Fuling group and model control group, 10 rats in each group.The Resina draconis. at 0.50, 0.25 and 0.12 g.kg-1 .d-1 was administered to the rats in the Resina draconis. high-, medium-, and low-dose groups, Guizhi Fuling capsule at 0. 25 g . kg-1 . d-1 to those in Guizhi Fuling group, and the same volume of 0. 5% sodium carboxyl methyl cellulose solution to those in model control group and blank control group. All treatments lasted for 3 weeks.At the end of the treatment, the rats were sacrificed;the volume and weight of ectopic tissue were measured. The blood viscosity was detected with blood rheometer.The concentration of estradiol and progesterone was detected using enzyme-linked immune sorbent assay.The matrix metalloprotein-2 expression in ectopic tissues was determined by RT-PCR. Results High, medium and low dose Resina draconis. significantly reduced the volume and weight of the ectopic tissue, and inhibited MMP-2 expression in ectopic tissues.High-, medium-dose Resina draconis. significantly reduced plasma cutting middle and low shear viscosity of the rats.But the Resina draconis. had no obvious effect on serum estradiol and progesterone concentration. Conclusion Resina draconis. reduces blood viscosity and inhibits MMP-2 expression, which can be used for the treatment of blood stasis type of endometriosis.

Objective To explore the preventive effects of different extracts from Aspidopterys obcordata on renal tubu-lar epithelial cells injury induced by sodium oxalate in vitro, and initially identify the effective part for treating urolithiasis. Methods The injury model of HK-2 cells induced by sodium oxalate was established to screen the active parts of Aspidopterys obcordata by testing the protective effects of different polarity extracts on HK-2 injury cells through MTT method. Results Dif-ferent extracts from Aspidopterys obcordata improved the activity of HK-2 injury cells, which were elevated to 86.17% and 95.42%by 0. 5 mg?mL-1 and 1 mg?mL-1 aqueous extract, respectively. And the activity reached to 93. 59% and 84. 77% by 0.5 mg?mL-1 and 1 mg?mL-1 50% alcohol extracts, reached above 81.56% by 95% alcohol extracts,all of which showed sig-nificant difference compared with the model group. The HK-2 cells viability were elevated to 82. 53% and 91. 58% by 0.5 mg?mL-1 and 1 mg?mL-1 95% alcohol extracted ethyl acetate parts,and increased to 77.24% and 87.22% by 0.5 mg?mL-1and 1 mg?mL-1 of 95% alcohol isolated n-butanol extracts, approached to 95.46% and 81.36% by 0.5 mg?mL-1 and 1 mg?mL-1 water extracts, all of which showed significant difference compared with the model control group. Conclusion The aqueous extracts and alcohol extracts from Aspidopterys obcordata have obvious preventive effects on HK-2 cells injury, among which the ethyl acetate extracts, n-butanol extracts and water extracts present the remarkable effects, which are supposed to be the active parts for inhibiting calcium oxalate stone formation in vitro.

Objective To study the film￣forming property of film￣forming materials in dragon blood ethanol solution, and provide the reference for screening of film￣forming materials for dragon blood plastics. Methods PVA124,PVA1788, PVP￣K30, EC, MC, PVB were chosen to study the film￣forming property of film￣forming materials based on solubility, formation time, and adhesion. Results PVA1788 and PVP￣K30 mixed in dragon blood ethanol solution in proportion of 4:1 showed good film￣forming property, and the membrane was smooth, uniform and easy to tear off.However, PVA124 and PVP￣K30 mixed in proportion of 4:1 precipitated in dragon blood ethanol solution.EC, MC and PVB were not easy to tear off the membrane. Conclusion PVA1788 and PVP￣K30 mixed in proportion of 4:1 and added with propylene glycol is the optimal choice of film￣forming material for dragon blood spray film.

OBJECTIVETo observe dynamic changes of blood lead concentration in rats with long-term toxicity test with Goupi Gao by the flame atomic absorption spectrometry, in order to provide reference for safe administration of Goupi Gao.

METHODThe rats were administered with Goupi Gao by high-dose (7 g x kg(-1)), medium-dose (3.5 g x kg(-1)), low-dose (1.75 g x kg(-1)) by external use for consecutively 90 days. Then, the blood samples were collected from the rats before the administration and at 10, 30, 45, 52, 60, 90 d after the administration respectively, as well as 16 d and 28 d after the drug withdrawal. The samples were dispelled with microwave digestion system and then were determined by graphite furnace atomic absorption spectrometry for blood lead levels.

RESULTAccording to methodological study, the standard curve regression equation in this experiment was A = 0.004 9X + 0. 017, r = 0.999 5, with the detection limit up to 0. 380 microg x L(-1). The RSD was 1.4% by precision checks. Blood lead level of mixed blood samples was 175.77 microg x L(-1), whose RSD was 6. 0%. Blood lead concentration gradually increased after low-dose and medium-dose administration to rats and became stable at the 10th day and the 30th day by high-dose. Dose is directly related to blood lead concentration. Meanwhile, the blood lead concentration decreases after the drug withdrawal.

CONCLUSIONThe method of determination in this experiment is so accurate and reliable that it can be used for the determination of blood lead. Long-term and high-dose external use of Goupi Gao can increase blood lead.

Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Female ; Lead ; blood ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry, Atomic ; methods ; Time Factors ; Toxicity Tests

Trigeminal neuralgia is a manifestation of orofacial neuropathic pain disorder,always deemed to be an insurmountable peak in the field of pain research and treatment.The pain is recurrent,abrupt in onset and termination similar to an electric shock or described as shooting.A poor quality of life has been attributed to trigeminal neuralgia,as the paroxysms of pain may be triggered by innocuous stimuli on the face or inside the oral cavity,such as talking,washing face,chewing and brushing teeth in daily life.The pathogenesis of trigeminal neuralgia has not been fully elucidated,although the microvascular compression in the trigeminal root entry zone is generally considered to be involved in the emergence and progression of the pain disorder.In addition,orofacial neuropathic pain restricted to one or more divisions of the trigeminal nerve might be secondary to peripheral nerve injury.Based on current hypotheses regarding the potential causes,a variety of animal models have been designed to simulate the pathogenesis of trigeminal neuralgia,including models of compression applied to the trigeminal nerve root or trigeminal ganglion,chronic peripheral nerve injury,peripheral inflammatory pain and center-induced pain.However,it has not yet been possible to determine which model can be perfectly employed to explain the mechanisms.The selection of appropriate animal models is of great significance for the study of trigeminal neuralgia.Therefore,it is necessary to discuss the characteristics of the animal models in terms of animal strains,materials,operation methods and behavior observation,in order to gain insight into the research progress in animal models of trigeminal neuralgia.In the future,animal models that closely resemble the features of human trigeminal neuralgia pathogenesis need to be developed,with the aim of making valuable contributions to the relevant basic and translational medical research.

Objective Retrospectively analysis on death cases related with aortic dissection provides support for further forensic investigation on such cases. Methods 44 Cases on aortic dissection are collected from the Forensic Medical Center in Kunming Medical University from 2002to2017.Statistic anaylisis was applied to each case's general information,clinical stage,clinical manifestation,risk factors,pathological type,rupture state,and direct case of death. Results The results showed that aortic dissection occurs moslty in middle age male who ranges from 40 to 49 years old. Most cases were aute aortic dissection, and the death rate was the highest in winter and the lowest in spring. Furthermore, pain is shown to be the most common symptom. Atherosclerosis is a critical factor that contributes to the occurance of aortic dissection. In general, type A aortic dissection occurs more frequently than type B. The rupture, located in ascending aorta, often is in a transverse shape. Cardiac tamponade is the primary cause of death.Conclusion Definite initial symptoms arises before aortic dissection rupture and always show many high-risk factors. It is beneficial to forensic identification of the relation between aortic dissection and injury or medical behavior should be considered carefully.

Objective:To analyze the characteristics of eyes with congenital optic disc pits (ODPs) through multimodal imaging.Methods:A cross-sectional study was conducted.Thirty-eight patients (38 eyes) diagnosed with congenital ODPs in the Second Hospital of Hebei Medical University from January 2009 to January 2020 were enrolled.A comprehensive summary analysis of the imaging results including fundus photography, spectral domain-optical coherence tomography (SD-OCT), infrared imaging, fundus autofluorescence (FAF), fluorescein fundus angiography (FFA) and indocyanine green angiography (ICGA) was performed.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The Second Hospital of Hebei Medical University (No.2021-P011). Written informed consent was obtained from each subject prior to any medical examination.Results:Among the 38 eyes, there were 32 eyes with ODPs located in or below the temporal side of optic disc, 4 eyes with ODPs located above the temporal side of optic disc, and 2 eyes with ODPs located at the center of optic disc, which were round or quasi-circular pale depression, and dark red eminences with clear or unclear boundaries between milk spots were found in 29 eyes with optical-disc macular degeneration (ODP-M) by fundus photography.SD-OCT examination showed that the structure of lamina cribrosa in the lesion area in all ODPs patients was incomplete, which presented a dark area with no tissue reflection, and the fissure led to the deep optic nerve.Fluid was found in the outer nuclear layer in all ODP-M patients, and there were 27 eyes with fluid in the inner nuclear layer, 13 eyes in the ganglion cell layer, and 4 eyes under the inner limiting membrane.Among the 29 eyes with ODP-M, there were 21 eyes with retinoschisis in outer layer, 27 eyes with neuroepithelial detachment.In the 27 eyes with neuroepithelial detachment, spot-like high reflection and reduced or disappeared ellipsoid band reflectance were seen above the neuroepithelium in 18 eyes.In infrared images, there were circular or quasi-circular low-reflection areas in the temporal side of the optic disc, and the lesion of ODP-M eyes presented low-reflection areas.FAF examination showed that in 27 eyes with ODP-M, there was a hypofluorescent region at the posterior pole consistent with the lesion range, among which, there was a granular or sheet-like hyperfluorescence at the center of the hypofluorescent region in 18 eyes.FFA showed that the optic disc depression in the arterial phase of patients was in a localized hypofluorescence state.During the venous phase, fluorescein dye extravasation along the temporal side of the optic disc could be found.A strong fluorescent arc with unclear boundaries at the temporal edge of the optic disc was formed in the late stage of angiography.Among the 29 eyes with ODP-M, the area of the macular lesion showed hyperfluorescence during the late stage of angiography in 27 eyes with neuroepithelial detachment, and no extension of dye toward the macula was found.ICGA showed that the optic disc depression of ODPs patients presented a localized hypofluorescence, and the lesion showed hyperfluorescence in 27 of the 29 ODP-M eyes with neuroepithelial detachment.Conclusions:Multimodal imaging can be helpful to realize the early diagnosis, etiology analysis of ODPs and make treatment plan.