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.

In large prospective studies, plasma fibrinogen levels have been shown to be an independent risk factor of vascular disease, including ischemic stroke. Elevated plasma fibrinogen in an individual could be due to the presence of predisposing genetic and/or environmental factors, such as smoking. Of the polymorphisms studies to date, the beta-fibrinogen-455 (beta-Fg-455) G-->A substitution in the 5' flanking region is associated with the most consistent difference in plasma fibrinogen levels in both case-control studies and in selected groups of healthy individuals. In order to further elucidate the role of the beta-Fg-455 G-->A substitution in determining fibrinogen levels and susceptibility to ischemic stroke in case-control population, including 104 individuals with verified ischemic stroke and 156 healthy individuals. Turbidimetriy assays were used to measure plasma fibrinogen levels of all samples. The beta-Fg-455 G-->A mutation was identified by the polymerase chain reaction followed by restriction enzyme digestion of the amplified DNA with HaeIII. The plasma fibrinogen level in patients with ischemic stroke [(3.51 +/- 1.09) g/L] was significantly higher than that in the control [(3.08 +/- 0.71) g/L] (P < 0.01). The A-allele is associated with elevated fibrinogen levels in both patients and controls. The plasma fibrinogen levels in controls with A-allele in elder people were higher than in younger people (P < 0.05). Those with A allele in males of ischemic stroke had significantly higher plasma fibrinogen levels in smokers than in non-smokers and ex-smokers (P < 0.05), but it was not significantly difference in subjects of GG genotype (P > 0.05). Our data demonstrates an association of the beta-Fg promoter A-455 allele with higher fibrinogen levels in the general population, and suggests that the A-allele may be a susceptible predictor of ischemic stroke, particularly in aging and smoking.

AIMTo investigate the effects of glycyrrhiza decoction on migrating myoelectric complex (MMC) and gastrointestinal hormone in small intestine in rats.

METHODSWe observed MMC cycle,phase Ill duration,fast wave numbers of phase III of MMC in one minute, fast wave numbers of one cluster in phase III of MMC of small intestine of glycyrrhiza group and control group rats with electrophysiology method, and immunohistochemistry to examine relative content of serotonin (5-HT), substance p(SP) and vasoactive intestinal polypeptide (VIP) in small intestinal chromophil (EC) and myenteric nerve plexus in small intestine of control group and glycyrrhiza group rats.

RESULTSCompared glycyrrhiza group with control group,we found that glycyrrhiza was able to decrease fast wave numbers in one minute and fast wave numbers in one cluster in phase III of MMC of small intestine (P < 0.05), and evidently extend small intestinal cycle of MMC (P < 0.05), it also shortened the phase III III duration (P < 0.05) or made the phase III of MMC absent. Compared glycyrrhiza group with control group it was indicated that content of 5-HT in small intestinal mucous membrane and myenteric nerve plexus was evidently decreased (P < 0.05), and content of SP in myenteric nerve plexus of small intestine of rats was evidently decreased (P < 0.05), and content of VIP in small intestine of rats was evidently increased (P < 0.05).

CONCLUSIONGlycyrrhiza is able to inhibit small intestinal motility, this inhibition is related with the amount of 5-HT, SP, VIP secreted by small intestinal mucous membrane of rats.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Electromyography ; Gastrointestinal Motility ; drug effects ; physiology ; Glycyrrhiza ; Intestine, Small ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Serotonin ; analysis ; Substance P ; analysis ; Vasoactive Intestinal Peptide ; analysis

OBJECTIVETo detect the effect of connective tissue growth factor (CTGF) on podocalyxin expression in mouse podocytes exposed to high glucose in vitro and explore the possible pathway involved.

METHODSThe expression vector carrying a small interfering RNA (siRNA) targeting CTGF was transfected into mouse podocytes cultured in the presence of 1 g/L glucose (normal control), 4.5 g/L glucose (high glucose group), 1 g/L glucose + 3.5 g/L mannitol (iso-osmolar control group). The changes in the protein expression levels of podocalyxin, CTGF and ERK1/2 in the cells in response to the treatments were investigated using Western blotting.

RESULTSHigh glucose exposure for 24 and 48 h resulted in significantly decreased expression of podocalyxin and increased CTGF in the podocytes (P<0.05). Phosphorylation of ERK1/2 occurred as early as 30 min after the exposure, and the activation was maintained till 24 h. Transfection of the cells with siRNA targeting CTGF significantly inhibited these changes.

CONCLUSIONCTGF is an important mediator of high glucose-induced podocyte damage and decreases the protein level of podocalyxin by the ERK1/2 pathway. CTGF-specific siRNA can alleviate high glucose-induced podocyte injury, suggesting its potential value in treatment of diabetic nephropathy.

Animals ; Cells, Cultured ; Connective Tissue Growth Factor ; metabolism ; Diabetic Nephropathies ; Glucose ; adverse effects ; MAP Kinase Signaling System ; drug effects ; Mice ; Podocytes ; cytology ; drug effects ; metabolism ; RNA, Small Interfering ; genetics ; Sialoglycoproteins ; metabolism