Objective: To study the origin, morphological structure, and functional regulation of osteoclast(OC) for further investigation on the mechanism and regulation of bone resorption. Methods: The OCs were isolated by two kinds of traditional method. Osteoclasts were isoclated from neonatal rat long bones. The cytochemistry was observed. The osteoclast-like cells (OLC) were derived from the mouse bone marrow cells in the presence of 1,25(OH)2VitD3 in vitro. Results: Both morphological and functional studies showed that the isolated cells shared some of the typical characteristics of osteoclasts, that is A. multinuclearity; B. developing spreading and pseudopodial activity when cultured on glass; C. high tartrate-resistant acid phosphatase (TRAP); D. resorption lacunae could be found when the cells were cocultured with devitalized bone slices and the number was increased as the time followed. OLC had the same histological and structural traits as the OCs by the former method. The concentration of Ca 2+ and acid phosphatase (ACP) increased gradually. Conclusion: Different kinds of method fit different experiments. The OC obtained by the first method has more activity of bone resorption. The OLC by the second method has more in quantity and can be used in the study of cell differentiation.

One hundred and sixty non-diabetic patients with cerebral ischemia of primary onset were enrolled in the study. The patients were divided into impaired glucose tolerance (IGT) group and normal glucose tolerance (NGT) group according to the oral glucose tolerance test (OGTT). Carotid atherosclerosis and carotid artery stenosis were examined with vascular Doppler ultrasonic detection. The incidence rates of carotid atherosclerosis in IGT group and NGT group were 87. 5% (70/80)and 63.8% (51/80)respectively (χ2= 12.240,P <0.001);the incidence rates of carotid stenosis in IGT group and NGT group were 63.8% (51/80) and 47. 5 % (38/80), respectively(χ2 =4. 279, P =0. 039);the degree of carotid artery stenosis in IGT group was higher than that in NGT group(χ2= 21.95, P < 0. 01). The results indicate that IGT may be related to atherosclerosis and stenosis of carotid artery in cerebral ischemic patients.

Objective To investigate the error monitoring function damages on the patients with schizo?phrenia ( SCH) . Methods A total of 32 patients with schizophrenia were compared with matched 34 health con?trols ( HC) on the error monitoring tasks which were compiled by E?Prime. Results The comparison between SCH group ((713.22±174.52)ms,( 491.14±170.29) ms,( 1060.31±130.84) ms,(8.28±12.55)time,( 8.00± 7.53)time respectively) and HC group ((560.73±156.94) ms,(395.62±188.03) ms,(989.85±104.33) ms, (2.97±4.13) times,(3.12±6.50) times) on the reaction time of choice,assessment,incongruent condition,the numbers of uncertain and the numbers of dropout were significant ( t=-3.737, P=0.000;t=-2.159, P=0.035;t=-2.426, P=0.018;t=-2.282, P=0.022;t=-2.824, P=0.006) . The SCH group and HC group did not signifi?cantly difference in Full Correct((124.72±23.74)/(131.74±21.96)times),Full Error((15.69±17.64)/(13.35± 18.63)times),Part Correct((6.83±10.40)/(4.21±7.03)times),Part Error((2.91±10.91)/(0.62±1.10)times) and Accuracy((0.831±0.161)/(0.874±0.159))(P>0.05).There was no significantly correlation among the course of disease,HAMA,HAMD and the error monitoring. Conclusion These results demonstrate that the error monitoring function damages on the patient with SCH may be involved in the dysfunction of anterior cingulate cortex.

OBJECTIVETo ascertain whether proanthocyanidins inhibit cell growth and migration by increasing let-7a expression in pancreatic cancer AsPC-1 cells.

METHODSThe proliferation rate, cell apoptosis rate and cell migration ability of AsPC-1 cells treated with proanthocyanidins were measured by MTT assay, Annexin V-FITC/PI staining, and Transwell migration assay, respectively. The expression of let-7a AsPC cells was detected by miRNA real-time RT-PCR after proanthocyanidins treatment. The changes in the biological behaviors of AsPC-1 cells were evaluated after transfection with let-7a mimics.

RESULTSCompared with the control group, proanthocyanidins treatment caused dose-dependent decrements of the proliferation rate and migration ability and increased the apoptosis rate in AsPC-1 cells. AsPC-1 cells with proanthocyanidins treatment showed increased expression of let-7a. Transfection with let-7a mimics resulted in obvious decreases in the cell growth rate and migration ability, and proanthocyanidins treatment significantly enhanced the inhibitory effect of let-7a mimics.

CONCLUSIONProanthocyanidins-induced cell growth and migration inhibition are partially mediated by up-regulation of let-7a expression in AsPC-1 cells.

Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Humans ; MicroRNAs ; metabolism ; Pancreatic Neoplasms ; pathology ; Proanthocyanidins ; chemistry ; Transfection ; Up-Regulation

OBJECTIVE: To explore the eff ect of grape seed proanthocyanidins extract (GSPE) on the growth of pancreatic cancer cells and the underlying mechanisms. METHODS: The pancreatic cancer AsPC-1 cells were cultured in vitro. The effects of GSPE on cell proliferation, apoptosis and migration were analyzed by MTT, Annexin V-FITC/PI and Transwell migration assay, respectively. The expression of miR-27a and FOXO1 in AsPC-1 cells was determined by real-time RT-PCR and Western blot, respectively. The miR-27a inhibitors were applied to verify the role of miR-27a in mediation of GSPE effects. RESULTS: GSPE inhibited cell growth in a dose-dependent manner. This inhibitory effect was significant when the dosage of GSPE was more than 50 μg/mL (P<0.05 vs control). GSPE also could induce apoptosis and inhibit cell migration. MiR-27a expression was notably down-regulated when the dosage of GSPE was 75 μg/mL (P<0.01 vs control). Compared with the control group, cell proliferation inhibition was significantly increased in the miR-27a inhibitor group, the GSPE group and the miR-27a inhibitor plus GSPE group (P<0.01), while cell migration was significantly decreased (P<0.01). Compared with the GSPE or the miR-27a inhibitor group, the growth and migration inhibitory effects in the miR-27a inhibitor plus GSPE group were more obviously (P<0.01). Both GSPE and miR-27a inhibitor alone could up-regulate FOXO1 expression. But these effects were more apparent when they are applied in combination. CONCLUSION GSPE inhibites AsPC-1 cells' growth and migration partly through down-regulation of miR-27a expression.
Apoptosis ; Cell Line, Tumor ; drug effects ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Grape Seed Extract ; pharmacology ; Humans ; MicroRNAs ; genetics ; metabolism ; Pancreatic Neoplasms ; pathology ; Proanthocyanidins ; pharmacology ; Up-Regulation

Objective To investigate the anti-inflammatory action of Celastrol in the ocular tissues of mice with ex-perimental autoimmune uveitis(EAU)and its effect on microglia polarization.Methods A total of 36 healthy B10.RⅢmice at 6-8 weeks of age were selected and randomly divided into the normal control group,EAU solvent control group and Celastrol intervention group,with 12 mice in each group.The interphotoreceptor retinoid-binding protein(IRBP)161-180 and Freund's complete adjuvant were mixed by thorough emulsification and injected subcutaneously into the bilateral thighs and tails of mice in the EAU solvent control group and the Celastrol intervention group with a total volume of 200 μL and 50 μg IRBP 161-180 in each mouse.On 7-14 days after immunization,mice in the Celastrol intervention group received a daily intraperitoneal injection of 0.5 mg·kg-1 Celastrol,and mice in the EAU solvent control group were injected with an equivalent dose of sterile Phosphate Buffered Saline solution.On the 14th day after immunization,the anterior segment of mice in each group was observed by slit-lamp microscope and Hematoxylin and Eosin(HE)staining of tissue sections was performed;the clinical and histopathological scores of mice in each group were obtained by reference to the Caspi grading standards;immunofluorescence staining was used to observe the activation of microglia in the eyes of mice;Western blot was used to detect the protein expression levels of inducible nitric oxide synthase(iNOS)and arginase-1(Argl)in the reti-na;quantitative real-time PCR was used to detect the relative mRNA expression of inflammatory factors in the retina,such as tumor necrosis factor(TNF)-α,interleukin(IL)-1β and IL-6.GraphPad Prism 9.0 was used for data analysis.Results On the 14th day after immunization,it was observed by the slit-lamp microscope that the anterior segment of mice in the EAU solvent control group was markedly congested with dilated iris blood vessels,corneal edema,and anterior chamber exudation;the inflammation in the anterior segment of mice in the Celastrol intervention group was markedly at-tenuated,and the iris blood vessels were seen to be mildly congested.Compared with the normal control group,the clini-cal scores of mice in the EAU solvent control group and the Celastrol intervention group were significantly elevated(both P＜0.05);the clinical scores of mice in the Celastrol intervention group were lower than those in the EAU solvent control group(P＜0.05).HE staining results showed that on the 14th day after immunization,mice in the EAU solvent control group showed severe retinal folds and detachment with diffuse infiltration of inflammatory cells,while mice in the Celastrol intervention group showed slight structural damage to the retina and a small amount of inflammatory cell infiltration.Com-pared with the normal control group,the histopathological scores of mice in the EAU solvent control group and the Celas-trol intervention group were significantly elevated(both P＜0.05);the histopathological scores of mice in the Celastrol in-tervention group were lower than those in the EAU solvent control group(P＜0.05).The intraocular Iba1+cell densities of mice in the normal control,EAU solvent control and Celastrol intervention groups were(1.00±0.12)％,(36.07± 4.57)％,and(1.83±0.36)％,respectively.Compared with the normal control group,the number of Iba1+cells in the eyes of mice in the EAU solvent control group and the Celastrol intervention group significantly increased(both P＜0.05);compared with the EAU solvent control group,the number of Iba1+cells in the eyes of mice in the Celastrol intervention group was significantly reduced(P＜0.05).Compared with the normal control group,the expression levels of iNOS and Arg1 proteins in the retinas of mice in the EAU solvent control group were significantly elevated(both P＜0.01);compared with the EAU solvent control group,the expression of iNOS protein in the retinas of mice in the Celastrol intervention group was significantly reduced(P＜0.01).Compared with the normal control group,the relative mRNA expressions of TNF-α.IL-1β,and IL-6 in the retinas of mice in the EAU solvent control group was significantly elevated(all P＜0.05);compared with the EAU solvent control group,the relative mRNA expressions of TNF-α,IL-1 β,and IL-6 in the retinas of mice in the Celastrol intervention group significantly decreased(all P＜0.05).Conclusion Celastrol inhibits Ml microglia activation and reduces the production of retinal inflammatory factors TNF-α,IL-1 β and IL-6 in EAU mice,thereby attenuating the in-flammatory reaction.

ObjectiveTo understand the prevalence of hookworm infection and its relevant behavioral factors in rural areas of Tiantai County， Zhejiang Province， and to provide scientific evidence for prevention and control of hookworm disease. MethodsBy using a stratified cluster random sampling strategy， local residents aged ≥3 years was divided into 5 districts according to geographical location； furthermore， those in one administrative village （surveillance site） were investigated in each district. Species of hookworm were identified by filter paper culture in vitro， and enterobius vermicularis eggs were detected by cellophane anal swab in children aged 3‒9 year. Risk factors were determined by questionnaire. ResultsA total of 1 013 residents were investigated in 5 surveillance sites. Thirty nine cases with hookworm infection were detected， with the total infection rate of 3.85% . All species detected were determined to be Necator americanus. The infection rate significantly differed across the towns （χ²=48.32， P<0.05）， with the highest rate in Nanping Town （10.95%） . It significantly differed by age groups （χ²=65.65， P<0.05）， with the highest rate in those aged >70 years （9.75%）. Furthermore， it decreased with educational background. It was significantly associated with fertilize with fresh manure （χ²=6.87， P<0.05） and barefoot labor （χ²=157.69， P<0.05）. ConclusionThe overall infection rate of hookworm in Tiantai County remains low. Dominant species of hookworm is hookworm Necator americanus. It is necessary to strengthen the advocacy of hookworm prevention and control knowledge， improve hygiene in work and life style， and increase self-protection awareness.

Objective To observe the effect of Zhuang Medicine Jib on spasm in hemiplegics after stroke. Methods From January, 2014 to December, 2015, 55 patients with spastic hemiplegia after stroke were divided randomly into treatment group (n=27) and control group (n=28). Both groups accepted routine rehabilitation and acupuncture, while the treatment group accepted Zhuang Medicine Jib in addition. They were assessed with modified Ashworth Scale (MAS), Barthel index (BI), Simpling Fugl-Meyer Assessment (FMA) before and four weeks after treatment. The side-effects within three months after treatment were recorded. Results The scores of MAS, BI and FMA improved in both groups after treatment (t>3.32, P<0.01), and improved more in the treatment group than in the control group (t>2.286, P<0.05). No side-effect was found in the follow-up. Conclusion Zhuang Medicine Jib can further relieve spasm after stroke, and improve the motor function and activities of daily living.