Background The pathogenic mechanism of thyroid associated ophthalmopathy (TAO) is still unclear,which is considered to be an autoimmune disease.It is confirmed that interleukin-17A (IL-17A) plays an important role in the occurrence and development of many autoimmune diseases.It is unclear that whether IL-17A participates in the pathogenesis of TAO.Objective This study was to explore whether IL-17A secreted by coculture system of peripheral blood mononuclear cells (PBMCs) and orbital fibroblasts (OFs) participates in the pathogenesis of TAO and its possible mechanism.Methods Periphery blood and orbital connective tissue were obtained from 12 patients with TAO and 8 patients who received prosthesis implantation for eyeball atrophy in Xiangya Hospital during April to December 2014.PBMCs were isolated by density gradient centrifugation,and OFs were cultured by explant culture method.The purity of T leukomonocyte in PBMCs was tested by flow cytometry,and OFs were identified by Giemsa staining and immunochemistry.OFs and PMBCs were incubated into 96-well plate in a 1:20 proportion to establish co-culture system.Different concentrations of phytagglutinin (PHA) (0,1.0,2.5,5.0,10.0 μg/ml) was added for 72 hours,and IL-6,IL-17A levels in the co-culture system supernatant and IL-17A receptor (IL-17RA) of the total cell membranes in the co-culture system were assayed by ELISA.The differences of IL-6,IL-17A,IL-17RA levels in co-culture system were compared between the TAO group and control group.Results The mean purity of T leukomonocyte in PBMCs was (81.10±0.21)％ in the TAO group and (80.05 ±0.38)％ in the control group respectively,with no significant difference between them(t =0.923,P＞0.05).Cultured OFs showed the positive response for Vimentin expression and Giemsa staining.After stimulated by 1.0 μg/ml PHA,the proliferation of both PBMCs and OFs were increased in the co-culture system.Apoptosis exist in PBMCs and the number of OFs decreased when PHA was higher than 1.0 μg/ml.The growth of PBMCs and OFs was faster in the TAO group than that in the control group in the same concentration of PHA.The contents of IL-6,IL-17A and IL-17RA in co-culture system were significantly different among various concentrations of PHA subgroups (IL-6:Fgroup =12.561,P=0.000;F ion =23.356,P =0.001.IL-17A:Fgroup =12.037,P =0.000;Fconcentration =19.206,P=0.000.IL-17RA:Fgroup =16.216,P=0.000;Fconcentraction =4.627,P=0.018).The production of IL-6,IL-17A and IL-17RA reached peak in both TAO group and the control group after 1.0 μg/ml PHA stimulated.However,the concentrations of IL-6,IL-17A and IL-17RA reduced with the increase of PHA concentration.The concentrations of IL-6,IL-17A and IL-17RA in co-culture system were significantly higher in the TAO group than those in the control group under the stimulation of the same concentration of PHA (all at P＜0.05).Conclusions The co-culture system of PBMCs and OFs stimulated with PHA can be the imitation of TAO pathogenesis in vitro,and PHA can amplify its immune reaction to imitate TAO pathogenic processes intuitively.The IL-6,IL-17A and IL-17RA secreted by PBMCs and induced by PHA are increased in TAO patients,implying that IL-17A participates in the pathogenesis of TAO through magnifying cellular immune response and inflammatory reaction.

Objective To investigate the feasibility of spatiotemporal image correlation(STIC)to assess left ventricular volume in normal fetuses.Methods The volume data acquisition of 162 normal fetuses ranging between 22 and 38 weeks of gestation was achieved by automatic sweep scan.The virtual organ computer-aided analysis(VOCAL Ⅱ)was then used to evaluate end-systolic and end-diastolic volumes of left ventricle and compared with that obtained by M-mode Teichholz formula and 2D biplane Simpson's method.Results Twenty-six fetuses with poor STIC image and 2 twin pregnancies were excluded,the study subjects consisted of 134 singleton fetuses.The highest concordance correlation(r =0.968,for EDV,r =0.956 for ESV;P =0.000)was noted between 2D biplane Simpson' s and STIC VOCAL Ⅱ values with a small bias(0.282 ml for EDV,0.117 ml for ESV).The Bland-Altman analysis showed that STIC VOCAL Ⅱ had the best agreement than M-mode Teichholz formula and 2D biplane Simpson's method.The result showed that there were significant difference between Mmode Teichholz formula and Simpson's method in EDV and ESV(P＜0.05).There were significant differences between M-mode Teichholz formula and STIC VOCAL Ⅱ(P＜0.05).There were no significant differences between STIC VOCAL Ⅱ and Simpson' s method in EDV and ESV.Conclusions There is a good agreement between left ventricular volumes measured either by STIC VOCAL Ⅱ or by 2D biplane Simpson's method.STIC is a novel technique that can be more accurate than conventional Methods in quantification of fetal left ventricular volumes.All the volume data acquisition is achieved by postprocessing facilities.STIC promises to become a new method for left ventricular volume quantification.

Objective To evaluate the clinical significance and cardiac function of fetuses with intracardiac hyperechogenic focus(ICEF) by fetal echocardiography. Methods Fetal echocardiograms were performed on 1291 fetuses for evaluation of fetal heart structures and rhythms. Echocardiographic characteristics of ICEF were investigated. Fetal systolic and diastolic functions of 399 fetuses with an isolated ICEF were compared with 128 normal fetuses using conventional two-dimensional, M-mode, pulsed Doppler echocardiography and myocardial Doppler tissue imaging(DTI). Results The prevalence of ICEF was 34. 7% (448/1291). Twenty-four(5.4%) cases were found with cardiovascular malformations. There was 1 (0.22%) case of ICEF fetus associated with chromosomal abnormality (trisomy 21 ). Comprehensive fetal echocardiographic data analysis showed no significant differences in heart dimensions, global cardiac functions,and myocardial performances in the two groups( P ＞0.05). Conclusions An isolated ICEF in the fetal heart is not associated with increased risk for chromosome abnormalities. Fetuses with ICEF do not indicate impaired left ventricular systolic or diastolic function, therefore having no hemodynamic significance.

BACKGROUND:Rat models of diabetes mellitus type 2 (T2DM) are usually induced by the combination of high-fat diet and low-dose streptozotocin, but their effects on the intervertebral disc have not yet been reported. Endplate sclerosis is an important factor contributing to intervertebral disc degeneration. OBJECTIVE:To evaluate whether the rat T2DM model induced by high-fat diet combined with low-dose streptozotocin is suitable for the study of T2DM related intervertebral disc degeneration. METHODS:Thirty-two 3-month-old female Sprague-Dawley rats were divided into four groups (n=8 per group), including sham, bilateral variectomy, DM, and bilateral variectomy plus DM groups, followed by subjected to bilateral ovariectomy and/or high-fat diet combined with low-dose streptozotocin, respectively. The blood glucose level, body mass and glucose tolerance were determined. The bone mineral density of the lumbar spine was measured after 8-week streptozotocin treatment. The L5-6 segments were removed and cut through midst sagittal plane after decalcification, and then underwent Von Gieson staining and histological degeneration scoring, and the disc height and endplate thickness were measured. RESULTS AND CONCLUSION:Fasting blood glucose and random blood glucose levels in the DM and bilateral variectomy plus DM groups were significantly higher than those in the other two groups, and the insulin sensitivity in the DM and bilateral variectomy plus DM groups were significantly lower than that in the other groups (P<0.05). L4-6 vertebral bone mineral density in the bilateral variectomy group was significantly lower than that in the sham group (P<0.05);L5-6 vertebral bone mineral density in the DM and bilateral variectomy plus DM groups was significantly lower than that in the sham group (P<0.05). L5-6 vertebral histological scores in the DM and bilateral variectomy plus DM groups were significantly higher than those in the other groups (P<0.05). Similar with the bilateral variectomy group, there were chondrometaplasia and mucoid degeneration of nucleus pulposus cells in the bilateral variectomy plus DM group, and the histological scores were significantly higher than those in the sham and DM groups (P<0.05). Compared with the sham group, the intervertebral disc height in the bilateral variectomy and bilateral variectomy plus DM groups was significantly decreased (P<0.05). While, there was no significant difference in the endplate thickness among groups. These results indicate the combination of high-fat diet and low-dose streptozotocin-induced rat T2DM models possess diabetic characteristics, but the rat intervertebral disc tissues show no significant differences from the normal ones;therefore, this model may be unsuitable for the study on T2DM-related intervertebral disc degeneration.

Objective To explore the expression of transforming growth factor ?1(TGF-?1)in the mucosa of guinea pigs with otitis media with effusion(OME)and the role of TGF-?1 in the development of OME.Methods 70 guinea pigs were randomly devided into 7 groups,each with 10 animals.OME was produced by injecting deactivated streptococcus pneumoniae into the typmpanic cavity of guinea pigs.The expression of TGF-?1 was examined by means of immunohistochemistry 6 h,1 d,3 d,7 d,14 d and 30 d after injection.Results TGF-?1 expression could be detected in the middle ear mucosa at 7 d after injection and reached maximum at 30 d.Conclusion TGF-?1 is expressed in the middle ear mucosa of the guinea pigs with OME induced by deactivated streptococcus pneumoniae,and this indicates that TGF-?1 may play a role in the chronic protraction of OME.

OBJECTIVE: To investigate the correlationship between the assessment of subjective and objective in patients with chronic(CRS) rhinosinusitis before and after FESS. METHOD: Seventy patients with CRS were objectively assessment of VAS and subjectively assessment including the Lund-Kennedy endoscopic scale and Lund-Mackay scale. All patients were followed up 1 year and repeated both the objective and subjective assessment. The correlationship among the different assessments were investigated and the scales of before and after FESS were compared. RESULT: There were positive correlation between the scale of VAS and Lund-Mackay(r = 0.866, P < 0.01), positive correlation between the scale of Lund-Kennedy and Lund-Mackay(r = 0.803, P < 0.01), and positive correlation between the scale of VAS and Lund-Kennedy (r = 0.912, P < 0.01) before the FESS. There were positive correlation between scale of VAS and Lund-Kennedy after 6 and 12 months of FESS with the r-value of 0.798 and 0.882 respectively. The scales of VAS including nasal obstruction, nasal discharge and head pain have positive correlation with the total scale of Lund-Kennedy with r-value of 0.691, 0.760 and 0.751 respectively. And the scales of VAS including hyposmia and general malaise have positive correlation with the total scale of Lund-Kennedy with r-value of 0.327, and 0.438 respectively. The scale of Lund-Mackay before has positive correlation with both the scale of Lund-Kennedy and VAS. CONCLUSION In patients with CRS, there were correlations among the scales of CT assessment, nasal endoscopic evaluation and VAS before FESS, and there were also correlations between the scales of CT assessment and nasal endoscopic evaluation after 6 and 12 months of FESS. It indicated that to evaluate the patients with CRS properly, the comprehensive assessment including subjective and CT and endoscopic scale should been applied in clinic.
Adolescent ; Adult ; Chronic Disease ; Endoscopy ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nose ; surgery ; Sinusitis ; surgery ; Treatment Outcome ; Young Adult

Objective To investigate the effect of long-term high-fat diet on insulin resistance and the morphology and function of islets in rats and the relationship between them.Methods Thirty normal male Wistar rats (8 weeks old) were divided into two groups and fed either with normal chow (NC,n=15),or high-caloric and high-fat diet (HF,n=15).Insulin resistance was assessed by euglycemic hyperinsulinemic clamp technique. The insulin secretory function of islets was evaluated by intravenous insulin releasing test.Morphological and quantitative analysis of pancreatic tissues was performed by double-label insulin and glucagon immunohistochemistry.Proinsulin mRNA was detected by RT-PCR.Results The glucose infusion rate (GIR) in HF rats was significandy lower than that in NC rats [(5.83?0.79)mg?kg~(-1)?min~(-1) vs (7.60?1.29)mg?kg~(-1)?min~(-1),P＜0.05].Immunohistochemistry showed that HF rats had larger islet size [(15168?1327)?m~2 vs (6264?1840)?m~2,P＜0.01] and significantly reduced insulin relative concentration of?cells[(-5.15?0.03) vs (-4.81?0.17),P＜0.01],as compared with NC rats.The islet relative?cell volume was decreased signifieandy (P＜0.01),whereas the relative?cell volume was increased (P＜0.01).So the ratio of?to?were lower in HF [(4.68?1.01) vs (11.84?3.82),P＜0.05].The peak of insulin secretion in intravenous insulin releasing test in HF was at 10 min,whereas that in NC rats was at 5 min.AUC (area under curve) 10-60 rain of insulin in HF was higher than that in NC rats [(152.51?34.53)mIU?L~(-1)?min~(-1) vs (86.40?21.21) mIU?L~(-1)?min~(-1),P＜0.01].There was no difference in proinsulin mRNA levels between two groups. Conclusion Long-term high-caloric and high-fat diet results in early impairment of islet morphology and function, as well as significant insulin resistance,suggesting that the compensation ability of islets has already been impaired in the early course of type 2 diabetes mellitus.

The purpose of this study was to investigate the molecular mechanisms whereby hydrogen sulfide (H2S) exerts the promoting effect on vascular endothelial cells migration. We used wound healing assay to study the effect of NaHS (H2S donor) on the migration ability of rhesus retinal pigment epithelial cell line, RF/6A cells, under normoxic conditions. Real-time PCR was used to measure hypoxia-inducible factor 1α (HIF-1α) mRNA level. Western blot was used to measure the expression of HIF-1α protein. The probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) was used to measure intracellular reactive oxygen species (ROS) level. The results showed that NaHS (10-100 μmol/L) could significantly promote RF/6A cells migration under normoxic conditions, and this effect could be inhibited by 50 µmol/L HIF-1 inhibitor, CdCl2. NaHS increased the protein level of HIF-1α in a dose- and time-dependent manner, and up-regulated the mRNA level of HIF-1α quickly and continuously. Moreover, NaHS could significantly decrease ROS levels in RF/6A cells under normoxic conditions. These results suggest HIF-1 may mediate the promoting effect of H2S on vascular endothelial cells migration under normoxic conditions. ROS, as an upstream regulator of HIF-1α, may be involved in the migration-promoting effect of H2S.
Animals ; Cell Line ; Cell Movement ; physiology ; Endothelial Cells ; cytology ; Hydrogen Sulfide ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Macaca mulatta ; RNA, Messenger ; genetics ; metabolism ; Reactive Oxygen Species ; metabolism ; Retinal Pigment Epithelium ; cytology ; Sulfides ; pharmacology

The case-based teaching method which is applied to the teaching of Science of Meridians, Collaterals and Acupoints is discussed in this paper, in which the typical cases such as the growth of the acupuncture-moxibustion eminent physicians, the application of acupoints by the eminent physicians and the experiences in the acupoint combination are integrated. The students are instructed to launch the clinical practical activity through establishing the clinical base of Science of Meridians, Collaterals and Acupoints, in association with the true cases. It is proved that the case-based teaching method can promote the training of the manipulation techniques of the students and the inheritance of the experiences of eminent physicians in higher education.
Acupuncture ; education ; Acupuncture Points ; Acupuncture Therapy ; methods ; Humans ; Meridians ; Students ; Teaching

Objective:To investigate the variation characteristics and influencing factors of HIV/AIDS subtypes in Wuxi city of Jiangsu Province from 2014 to 2016.Methods:HIV/AIDS population in Wuxi city in 2014 was selected as the research object, and the HIV molecular epidemiology and follow-up study were carried out. Collect epidemiological information, extract DNA from blood samples, amplify pol gene fragment by nest-PCR and sequence, use ChromasPro 1.6 software and MEGA 7.0 software to construct the HIV-1 sequence database, and use FastTree2.1.10 software to construct the phylogenetic tree to confirm the subtype; in 2016, the same population was followed up, and the HIV subtype variation was analyzed, and the influencing factors of subtype variation were explored by multivariate logistic regression. Results:A total of 612 HIV/AIDS cases in 2014 and 2016 were collected. The age of the subjects was mainly 30 years old or above (85.46%, 523/612), and the proportion of people over 50 years old was higher (228/612, 37.25%). The main route of transmission was homosexuality, accounting for 49.67%. A total of 1224 samples were detected and CRF01 _ AE、CRF07_ BC、B、CRF08_ BC、CRF67_ 01B、CRF55_ 01B、CRF68_ 01B, 7 subtypes of HIV-1 and 5 unique recombinant types (URFs) was detected. CRF01_ AE and CRF07_ BC was still the main genotype in Wuxi, Jiangsu Province, accounting for 66.75%. There were 29 cases (3.56%) of URFs recombinant strains. During 2014-2016, the variation rate of subtypes was 14.63%, and the most common variation was CRF01_ AE changes to CRF07_ BC(13.95%). Marital status (OR=0.363, 95% CI: 0.137-0.964) and baseline CD4 level (OR=0.414, 95% CI: 0.192-0.891) were associated with subtype variation.Conclusions:The HIV-1 subtypes of HIV/AIDS patients in Wuxi city are diverse and complex, the proportion of recombinant subtypes is rising, the URFs that are difficult to determine the genotype increase significantly, and the variation rate of HIV-1 subtypes among HIV/AIDS infected people is high. It is necessary to strengthen the monitoring of HIV-1 subtypes.