Objective To compare the efficacy of Graves disease animal models induced by thyroid stimulating hormone receptor (TSHR) plasmid DNA (pcDNA3.1-TSHR) and by TSHR A subunit recombinant adenovirus(Ad-TSHR289),and to investigate the influence of duration for preparing animal model induced by Ad-TSHR289 on Graves hyperthyroidism and its related indices.Methods The plasmid group and the adenovirus group were set up respectively.The plasmid group:21 female BALB/c mice were randomly divided into model group (n =12) and control group (n =9).The model group were injected intradermally with pcDNA3.1-TSHR 50 μg,once every 3 weeks,totally 3 times.Then 4 weeks after the last immunization,the mice were euthanized to obtain blood for testing TSHR antibody (TRAb),total T4,and thyroid tissue for histological examination.The controls were injected with the same dose of pcDNA3.1 in the same way.The adenovirus group:52 female BALB/c mice were divided into 10-week model group (n =8),14-week model group (n =10) and 18-week model group (n =8),and the respective controls (n =8,n =10,n =8) were set up.All model groups were injected intramuscularly with Ad-TSHR289,three times at three weekly intervals.Then the mice were euthanized at 4,8 and 12 weeks to test TRAb,total T4 level and to observe the change of thyroid histology.The controls were treated with the same dose of Ad-lacz in the same way.Another 8 mice were scheduled to test the dynamic variation of TRAb before and after the 3 times immunization.Results In the plasmid model group,only two of 12 mice developed weak antibody responses against TSHR,and no elevated total T4 level and no hyperplasia changes of thyroid were observed.In the 10-week model group,all mice had high level TRAb [(807.65 ± 136.33)U/L,Six-eighths mice had hyperthyroidism exhibited hyperplasia changes.In the 14-week model group,the TRAb level [(650.12 ± 192.88) U/L]and the incidence of hyperthyroidism (3/10) were lower than those in 10-week group.Histologically,the degree of thyroid hyperplasia lightened to a small extent,but its positive rate did not decline.In the 18-week model group,only 2 of 8 mice displayed slightly elevated TRAb level,and no mice showed increased total T4 level.Additionally,thyroid tissues of 2 mice were mildly abnormal.Compared with the model groups at different time,the change of antibody levels of the mice for TRAb dynamic observation exhibited the similar trend.Conclusions Being good at repeatability and high incidence of hyperthyroidism,the animal model of Graves disease induced by Ad-TSHR289 is still an ideal research tool presently.The duration of model ean be maintained 18 weeks,and 10 weeks is the best period to snstain characteristic of Graves disease.

Objective To understand the genetic characteristics of Enterovirus 71 ( EVT1 ) circu-lating strains of Guangdong province in 2008. Methods We isolated an EV71 strain from the fatal case of the hand, foot and mouth disease during an epidemic of Guangdong in 2008. Its complete genome was se-quenced and analyzed comparatively. Results The results showed that the full length of EV71 GDFS-3 ge-nome( not including poly A tail ) is 7405 bp. No insertion or deletion is detected in the coding region. There are several insertions and deletions in 5'and 3'UTR. Phylogenetic analysis of GDFS-3 and reference strains showed GDFS-3 strain shares the highest nueleotide homology with TW984 strain(96.0% ) but low homology with SIN5865, MS and BrCr( about 81.0% ). GDFS-3 strain also shares the highest amino acid homology with TW984 strain(99.0% ). It clustered with reference strains of CA subgenotype in the phylogenetie tree. The nucleotide identity with CA reference strains is 91.0% -95.0%. Conclusion The phylogenetic analysis based on the entire genome demonstrates that GDFS-3 strain has the nearest genetic relationship with TW984 strains ( isolated in 2004). GDFS-3 may belong to the same subgenogroup ( CA ) with Taiwan predominant strains. Otherwise,Some mutations in 5'UTR of EV71 may play the very important role in heightened viru-lence.

Objective: To investigate the chemical constituents in the whole plant of Lycopodium japonicum, native to Suizhou city, Hubei province for the first time. Methods: The compounds were isolated and purified by a combination of silica gel, ODS, and Sephadex LH-20 column chromatography and prep-HPLC. Their chemical structures were established by their physicochemical properties and spectral data. Results: Eight pentacyclic triterpenoids have been isolated and characterized from the methanol extract of the whole plant, they are 21-episerratenediol (1), serratenediol (2), α-onocerin (3), serrat-14-en-3β, 21β, 24-triol (4), 3β, 21α- dihydroxyserrat-14-en-16-one (5), lycoclavanol (6), lyclaninol (7), and 3α, 21β, 24-trihydroxyserrat-14-en-16-one (8). Conclusion: Compounds 1-8 are all serretene triterpenoids, the characteristic chemical constituents in the plants of genus Lycopodium L. Compound 5 is isolated from this plant for the first time.

Objective: To investigate the anti-inflammatory chemical constituents from the roots of Clerodendrum trichotomum. Methods: The compounds were isolated and purified by silica gel, Sephadex LH-20 column chromatographies, as well as prep-HPLC; Their structures were characterized on the basis of physicochemical properties and their spectral analysis; An in vitro anti-inflammatory method was applied to determine their activities against interleukin-8 (IL-8) production. Results: Four 24-ethylcholestane derivatives have been identified as 24-ethyl-7-oxocholesta-5, 22(E), 25-trien-3β-ol (1), decortinone (2), 22-dehydroclerosterol (3), and clerosterol (4). Conclusion: Coumpounds 1-3 are isolated from the roots of C. trichotomum for the first time. The absolute configuration of C-24 of compound 1 is determined to be S. In an in vitro anti-inflammatory assay, compound 1 is found to show a remarkable inhibition on IL-8 production induced by interleukin-1β (IL-1β) in HT-29 cells.

BACKGROUND:There is a lack of the research concerning the biocompatibility of nano-hydroxyapatite/polyphenylene sulfide (nHA/PPS) composites.OBJECTIVE:To evaluate the in vivo biocompatibility of nHA/PPS composites based on the completed research in vitro.METHODS:Systemic toxicity test:Sprague-Dawley rats were given the intraperitoneal injection of nHA/PPS extract or normal saline.The general situation,body mass and the histological changes of the liver and kidney were observed at 72 hours after injection.Delayed type hypersensitivity test:nHA/PPS extract or normal saline was injected subcutaneously into the back of the rats.Afterwards,skin irritation symptoms were observed at 72 hours.Local reaction experiment:nHA/PPS composites and polyethylene were respectively implanted into the back of the rats.The pathological changes of the implanted materials and their surrounding tissues were observed at 15 and 30 days after implantation.RESULTS AND CONCLUSION:(1) The rats were in good situation after nHA/PPS injection;the body mass increased steadily,which showed no significant difference from the control group (P ＜ 0.05);the morphology and color of the liver and kidney were normal,and the systemic toxicity of the composite materials was normal according to the degree of toxicity classification.(2) There were no obvious skin irritation symptoms after the subcutaneous injection of nHA/PPS composites,and the primary irritation index was less than 0.4,suggesting a low hypersensitivity.After implantation of nHA/PPS composites,there was no obvious degradation,absorption and rejection,and both the degree of inflammatory reaction (15 days ≤ level Ⅲ,30 days ≤ level Ⅱ) and the thickness of fibrous capsule (15 days ≤ level Ⅲ,30 days ≤ level Ⅱ) revealed the good biocompatibility of the composites.These results suggest that the nHA/PPS composites hold an excellent biocompatibility in vivo.

OBJECTIVETo compare the effect of different solvents such as water, ethanol, methanol, ethyl acetate and petroleum ether on extraction of 10 effective components from Ligusticum chuanxiong and component characteristics of corresponding extracts.

METHODUltrasonic assisted solvent extraction and high performance liquid chromatography quantitative analysis were adopted to determine effective components. CAPCELL PAK C18 column (4.6 mm x 150 mm, 5 microm) was adopted. The mobile phase was methanol-0. 5% HAc for gradient elute. The detection wavelength was 280 nm. The column temperature was 30 degrees C. The flow rate was 0. 7 mL x min(-1). The sample size was 10 microL.

RESULTMethanol or ethanol showed no significant difference in extraction of ferulic acid, hydroxyphthalide, alkylphthalide and diligustilide. Ethyl acetate displayed relatively low extraction ratios in hydroxyphthalide and ferulic acid. Water and petroleum ether showed relatively low extraction ratios in all four effective components, and water extracts showed different component characteristics.

CONCLUSIONEthanol and methanol are the most suitable solvents to extract four effective components from L. chuanxiong.

Drugs, Chinese Herbal ; chemistry ; Solvents ; chemistry

Objective:To study the value of ultrasound-guided endoscopic retrograde appendicitis therapy in children with uncomplicated appendicitis.Methods:This study was a single center, retrospective study, including all electronic cases of appendicitis diagnosed clinically in Department of Pediatrics, the Second Affiliated Hospital of Air Force Military Medical University from October 2018 to October 2020 and received ultrasound-guided endoscopic retrograde appendicitis therapy.The clinical features, treatment and prognosis of the children were retrospectively analyzed.Results:A total of 152 electronic cases were included, there were 77 males and 75 females, aged(6.84±3.09) years.All the 152 children were treated with ultrasound-guided endoscopic retrograde appendicitis therapy.Intubation success rate and clinical success rate was 98.03%(149/152 cases)and 97.99%(146/149 cases), respectively.The median time of endoscopic therapy was 42.50 (31.00, 56.00) minutes.Mean postoperative hospital stay was (2.81 ±1.41) days, and the mean total hospital stay was (4.19 ±1.71) days.A total of 139 patients were followed up with a median follow-up time of 5 (1, 26) months.During the follow-up, the recurrence rate was 7.19%(10/139 cases), and the median time of recurrence was 2 (1, 3) months.Conclusions:Ultrasound-guided endoscopic retrograde appendicitis therapy had high effective rate and low recurrence rate in children with uncomplicated appendicitis, preserved the physiological function of appendix and avoided radiation damage.It can be used as a safe and effective treatment for acute and chronic uncomplicated appendicitis in children.

An about 700 bp DNA fragment was amplified from genome DNA of S. aureus TSTw by PCR. This fragment was cloned into pGEM-7Zf(+) and the recombinant plasmid was transformed into E. coli DH5 alpha. The sequencing result of the recombinant plasmid demonstrated that it contains seb gene with 717 bp (without signal encoding region of 81 bp) which has the same nucleotide sequence as described in literature. The seb gene was cloned into expression vector 7ZTS and was transformed into E. coli JM109 (DE3). The expression level of SEB was as high as 33.3% of the cell total proteins.
Cloning, Molecular ; Enterotoxins ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Engineering ; Recombinant Proteins ; biosynthesis

The rare codons of a fragment in staphylococcal enterotoxin A gene were turned into the most high usage frequency codons in E. coli by overlap PCR technique. Genes of sea and seam were cloned into 7ZTS expression vector and transformed into JM109(DE3), respectively. The result shows that expression level of sea gene was very low, but the expression level of seam was as high as 15% of total cell proteins. The expression product shows activity of antitumor in vivo.
Animals ; Base Sequence ; Codon ; genetics ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; Gene Expression Regulation, Bacterial ; Male ; Mice ; Molecular Sequence Data ; Neoplasms, Experimental ; drug therapy ; pathology ; Point Mutation ; Recombinant Proteins ; genetics ; metabolism ; pharmacology

OBJECTIVETo study the effect of arctiin on mouse podocyte epithelial-mesenchymal transition (EMT) induced by advanced oxidation protein products (AOPP).

METHODSMouse podocytes were stimulated by 200 µg/ml AOPP for 24 h in the presence of 50, 100, 200, and 400 µmol/L arctiin. The expressions of α-smooth muscle actin, Grp78 and CHOP were detected using Western blotting.

RESULTThe expressions of α-SMA, Grp78 and CHOP were inhibited by arctiin, showing a dose-dependent effect within a given range of arctiin concentration.

CONCLUSIONAOPP causes endoplasmic reticulum stress to induce EMT of mouse podocytes, and arctiin can decrease EMT by alleviating the stress. This finding sheds light on a new scope of research of renal fibrosis.

Actins ; metabolism ; Advanced Oxidation Protein Products ; adverse effects ; Animals ; Cell Line ; Endoplasmic Reticulum Stress ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Furans ; pharmacology ; Glucosides ; pharmacology ; Heat-Shock Proteins ; metabolism ; Mice ; Podocytes ; metabolism ; pathology ; Transcription Factor CHOP ; metabolism