Background Normal ultrastructure is the anatomical basis of retinal pigment epithelial(RPE) cells to perform normal physiological function.At present the precipitation method is often used to detect the ultrastructure of RPE cells with transmission electron microscopy(TEM).Objective The aim of this study was to explore a simple and feasible approach to examine the ultrastructure of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells.Methods hESCs were induced and differentiated into RPE cells by the spontaneous differentiation method,and the expressions of microphthalmia associated transcription factor MITF and paired-box gene 6 (PAX6),specific protein of RPE cells,in the cells were detected by immunofluorescence assay.hESC-RPE cells were inoculated into Transwell filter,and the ultrastructure of the cell sheet was examined under the TEM.Then the ultrastructure of the cell sheet specimens was compared with those of hESC-RPE cells from cell precipitation and RPE cell specimens of 90-day-old Long Evans rats.Results MITF and PAX6 were positively expressed in hESC-RPE cells.The normal ultrastructure were visible in the RPE cells of rats under the TEM,including apical microvilli,polarized melanin granules,cellular nucleus,basement membrane and intercellular junctions,and the ultrastructure of hESC-RPE cell sheet on Transwell was similar to the RPE cells in rats.However,only scatter melanin granules,nonpolar nucleus and scanty microvilli were observed under the TEM in the hESC-RPE cells by cell precipitation method.Conclusions Without digestion process,hESC-RPE cell sheet on Transwell can retain the normal ultrastructure of hESC-RPE cells under the TEM,with a more simple and reliable advantage.

Objective To explore the feasibility of imaging and treatment of cervical cancer xenograft model using 131I mediated by hNIS gene transfection. Methods The cervical cancer xenograft models were established with Hela-NIS( +) cells and Hela cells, respectively. Five Hela-NIS( +) xenograft models and five Hela xenograft models were dynamically imaged at 0.5, 1, 2, 4, 8, 16 and 20 h postinjection of 131I(7.4 MBq). Five Hela-NIS( +) xenograft models were imaged at 0. 5,1,2,4,8,16, 20 and 25 h postinjection of 99TcmO4-(11.1 MBq). Twenty Hela-NIS( +) cervical cancer xenograft models were randomly divided into four groups: Three 131I treating groups and one control group. The therapeutic effects of 131I at threelevels (74,111,148 MBq) were investigated following intraperitoneal injection. Results Hela-NIS( +)human cervical cancer xenografts were established successfully in nude mice. The Hela-NIS( +) xenografts significantly accumulated radioactivity after intraperitoneal injection of 131I, and the radioactivity was persistently present until 20 h postinjection, but Hela xenografts had no radioactive accumulation. The T/B value of the Hela-NIS( +) xenografts reached 17.34 at 8 h postinjection. The imaging with 99TcmO4- showed that the radioactivity was persistently present in Hela-NIS( +) xenografts for almost 25 h. The Hela-NIS( +)xenografts shrinked after 131I treatment. The inhibition ratios of tumor growth in 111 MBq and 148 MBq groups were both significantly higher than that of 74 MBq group (t: 2.74-5.75, P ＜0.05). Conclusions Hela-NIS( +) cervical cancer xenografts in nude mice could persistently accumulate 131I and 99TcmO4- and could be treated successfully with 131 I. 131 I treatment mediated by hNIS gene transfection could be a promising cancer treatment method.

Objective To investigate the influencing factors for knowledge,attitude and practice(KAP)of the Three-manager mode about hierarchical medical system of Chronic diseases of the community residents in Xiamen,and provide references for the implementation of hierarchical medical system.Methods Multi-stage random sampling method was adopted,and 400 residents were randomly sampled from 5 communities of Xiamen for questionnaire survey.The survey features the dependent variable of the community residents′ knowing and joining-in the Three-manager mode,the internal factors(social demographic characteristics),and the external factors(medical expenditure and healthcare insurance).On such basis,four logistic regression models were built for analysis of such influencing factors.Results 41.10% of the surveyed were aware of the Three-manager mode,and 39.71% of the hypertension/diabetes patients among them joined in the hypertension network or diabetes network.Age,education,medical expenditure and referral experiences were significant influencing factors for KAP level of the Three-manager mode,while the residents of older age,higher education and referral experiences were high in their KAP;those of lower per capita monthly income,hypertension or diabetes,with coverage of medical insurance and satisfied with medical consultation in community were high in their awareness of Three-manager mode.Conclusions The KAP level of Three-manager mode of the community residents in Xiamen is acceptable,yet expected of further improvement.Room of further improvement includes enhancement of synergy of the Three-manager mode,enhanced motivation of government financial support and medical insurance,better service functions of community institutions,and especially the construction of hierarchical medical system.

Objective To investigate the relationship between cow's milk protein allergy(CMPA)and gastroesophageal reflux disease(GERD)and the prognosis of GERD combined with CMPA.Methods Fifty patients(24 boys and 26 girls)with GERD were enrolled in this study from January 2015 to June 2016 at Department of Pediatrics,Tangdu Hospital of the Fourth Military Medical University.All children were treated with serum milk protein soluble IgE(sIgE)and milk protein avoidance test,and those with positive results of children's milk protein by provocation test and those with milk serum protein sIgE negative by milk protein provocation tests were diagnosed as CMPA children with GERD according to the CMPA diagnostic criteria and received diet therapy for 1 month and then their blood eosinophil ratio and 24-hour esophageal pH were monitored.Results Twenty-three cases(46％)of 50 children with GERD were diagnosed as CMPA.There was significant difference in clinical symptoms between GERD group and GERD combined with CMPA group(x2=22.78,P<0.05),but there existed cross-symptoms among individual patients,so clinical accurate diagnosis turned out to be difficult.There was no significant difference in family history of allergy between GERD group and GERD combined with CMPA group(x2=3.19,P>0.05).For children with GERD combined with CMPA,the patients received dietary treatment for 1 month.There was significant improvement in vomiting,runny nose/wheezing/cough and diarrhea(P<0.05).However,because the treatment of eczema was long and it could relapse easily,there was no significant change after 1 month of therapy(P>0.05).The proportions of blood eosinophils were decreased after treatment compared with those before treatment [(2.7±1.8)％vs.(8.2±2.7)％,t=10.006,P<0.01].The results of 5 children's 24-hour esophageal pH monitoring showed that the reflux index and the number of acid GERD episodes were lower than before,and the difference was all statistically significant before and after(all P<0.05).Conclusions The occurrence of GERD in infants is partly related to CMPA,and the treatment of CMPA can relieve the clinical symptoms of GERD.

The purpose of this paper is to clarify the structure-activity relationship of anti-tumor activity of diosgenin derivatives in vitro. Study has found that diosgenin can inhibit the reproduction of tumor cells by inducing apoptosis and the main target spot of this effect is Bcl-2. Based on the characteristics of pharmacophoric points' of the three-dimensional pharmacophore for Bcl-2 inhibitors, we have docked lots of diosgenin derivatives with Bcl-2, then synthesized 31 compounds of them, finally assessed the anti-tumor activity of the diosgenin derivatives in vitro against A375, A549, HepG-2 and K562. Preliminary studies of SAR have indicated that the aliphatic esters, and aromatic esters of diosgenin without F ring have no anti-tumor activity in vitro. The triazole bromides of diosgenin all achieve fairly good anti-tumor activity in vitro, and those with larger hydrophobic group have the better activity. The stronger is the hydrogen bonding interaction and dipole-dipole interaction of the heterocyclic of diosgenin and diosgenin without F ring and the acid ester of diosgenin without F ring, the better is the activity of derivatives.
Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Diosgenin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; Structure-Activity Relationship

OBJECTIVETo compare the efficacy of decollement, combined tumescent liposuction and curettage, Botox injection in the treatment of osmidrosis.

METHODSFrom September 2006 to April 2009, 350 patients with osmidrosis were treated by decollement method in 180 cases, curettage in 120 cases,and Botox injection in 50 cases. The therapeutic effect and complications were retrospectively analyzed.

RESULTSThe patients were followed up for 3-24 months. The cure rate was 90.6%, 84.8% and 84.2% in decollement, curettage and Botox groups, respectively. The effective rate was 100% in all groups.

CONCLUSIONSAll the three methods are effective for the treatment of osmidrosis. The odor symptom can be improved a lot with decollement method. Considering the complication, downtime and cosmetic results, Botox injection is suitable for patients with minor or median odor, while combined liposuction and curettage is selected for patients with median to severe odor.

Axilla ; surgery ; Botulinum Toxins, Type A ; administration & dosage ; therapeutic use ; Curettage ; methods ; Female ; Follow-Up Studies ; Humans ; Hyperhidrosis ; surgery ; Lipectomy ; methods ; Male ; Retrospective Studies ; Treatment Outcome

Objective To retrospectively assess diagnostic accuracy of magnetic resonance imaging (MRI)in preoperative assessment of local extent of breast ductal carcinoma in situ(DCIS)and DCIS with small invasive foci,compared with the mammography and ultrasonography(US)imagings.Methods Results of MRI,mammography,and US imaging from 17 consecutive women with known breast DCIS and DCIS with invasive foci were analyzed,and then compared with pathologic examination.Results(1) Fourteen lesions showed enhancement on dynamic breast MRI,of which 11 lesions were no-mass-like enhancement.Six of 11 lesions appeared segmental enhancement,and 2 were regional enhancement.Ductal and multiple focal areas enhancement were 1 case respectively.Symmetric diffuse enhancement in bilateral breast was showed in 1 patient.Ductal dilation was visible in ipsilateral breast on pre-contrast MRI in 2 cases,which manifested bloody nipple discharge in clinical examination,and duct enhanced on post- contrast imaging in one of them.Two lesions appeared mass enhancement with irregular shape and homogeneous signal.Linear enhancement surrounding the oval homogeneous mass with smooth margin found in 1 case.(2)Thirteen of 17 patients underwent bilateral mammography.There were various findings in mammograms,including microcalcifications(6 cases),normal mammograms(2 cases),calcifications with other appearance(2 cases),and non-calcification abnormity(3 cases).In 8 lesions with calcifications, 5 were noted higher probability of malignancy calcifications and 3 intermediate concern calcifications. Calcifications distributed clustered(5 cases),regional(2 cases)and diffuse(1 case)shape.(3)Sixteen of 17 patients were performed breast US examination.Eleven lesions,which were correct diagnosed, appeared higher echo spots within irregular lower echo area.One lesion diagnosed benign and 4 were negative on US examination.(4)Regarded the size measured on pathologic examination as golden standard, accordance of lesion extent was 13 of 17 case in MRI,7 of 13 in mammography,and 7 of 16 in US.There were 2 lesions overestimation of extent in MRI,3 in mammography,and 2 in US.Underestimation of extent showed 1 case and 3 cases in mammography and US imaging respectively.The difference was not significant (P = 0.161).Conclusion The MR imaging features of DCIS and DCIS with small invasive loci were characteristic.The combination of MRI and mammography could improve diagnostic accuracy.

OBJECTIVETo investigate the social mental state of drug addicts in a compulsive drug abuse treatment center; evaluate the effectiveness of integrated program for the prevention of abuse relapse and improvement of drug addicts' psychological health.

METHODSThe study subjects were addicts from the Wuhan Compulsive Drug Abuse Treatment Center between October 2003 and June 2004, who satisfied the inclusion criteria. A non-randomized control-intervention study design was adopted. Volunteers willing to take part in intervention were put into the intervention group with their full awareness and willingness to prevent drug abuse relapse. The control group was composed of the addicts who were willing to prevent relapse and to be followed up after their discharge.

RESULTSThe effectiveness of the integrated intervention program in promoting addicts' psychological health: before the intervention, the scores of Self-Rating Anxiety Scale (SAS), the positive and negative dimensionalities of Simple Coping Style Questionnaire (SCSQ) and Chinese Perceived Stress Scales (CPSS) had no significant differences between the intervention group and the control group. After the intervention, except that the SCSQ's positive dimensionality in the intervention group was significantly higher than that in the control group, other indices in the intervention group were lower. Before and after the intervention, the psychological health level in both the groups was lower than that in the normal population; there were significant differences between addicts and normal subjects in regards with all of the indices above.

CONCLUSIONDrug abuse was associated closely with addicts' social mental factors. The integrated intervention program can alleviate anxiety and stress, reduce co-morbid mental disorders and effectively improve their coping style. In conclusion, the program can promote addicts' psychological health significantly.

Adaptation, Psychological ; Adult ; Case-Control Studies ; Female ; Humans ; Male ; Mental Health ; Program Evaluation ; Stress, Psychological ; Substance-Related Disorders ; psychology ; therapy

OBJECTIVETo investigate the role of the expression of ephrinB2 and EphB4 in non-small cell lung cancer (NSCLC), and their relationship with multi-slice spiral CT pulmonary perfusion imaging.

METHODSThirty-one nodules with pathologically proven NSCLC underwent CT perfusion scan. The perfusion parameters including blood flow (BF), blood volume (BV), peak enhancement image (PEI) were collected. The expression of ephrinB2 and EphB4 in tumor cells and interstitial vasculature were detected by immunohistochemistry. Correlation analysis and trend test were used to assess the relationship between ephrinB2/EphB4 expression and clinicopathological features, and between ephrinB2/EphB4 expression and perfusion parameters.

RESULTSPositive expression of ephrinB2 and EphB4 in the NSCLC group was 83.9% and 71.0%, respectively, significantly higher than that in the internal control group (P < 0.01). The expression of ephrinB2 and EphB4 was consistently in tumor parenchyma but differently in tumor vessels. The expressions of ephrinB2 and EphB4 were positively correlated with lymphatic metastasis (P < 0.05). The expression of EphB4 was negatively correlated with blood flow (BF) and blood volume (BV), respectively (P < 0.05). There was a significant positive correlation between ephrinB2 expression and BF (r = 0.516, P = 0.003), and a positive correlation between ephrinB2 expression and BV (r = 0.448, P = 0.013). The expressions of ephrinB2 and EphB4 were not correlated with PEI (P > 0.05). The values of BF and BV in the high and moderate EphB4 expression groups were significantly decreased compared with that in the negative group (P < 0.01). The value of BF in the high ephrinB2 expression group was significantly increased compared with that in the moderately positive group and negative group (P < 0.01). The value of BV in the high ephrinB2 expression group was significantly increased compared with that in the negative group (P < 0.01).

CONCLUSIONThe CT pulmonary perfusion imaging reflects the density difference of blood vessels with functional lumen, and such difference also depends on the quantity and quality of vasculature with functional lumen.

Adult ; Aged ; Blood Volume ; Carcinoma, Non-Small-Cell Lung ; blood supply ; diagnostic imaging ; metabolism ; pathology ; physiopathology ; Ephrin-B2 ; metabolism ; Female ; Humans ; Lung Neoplasms ; blood supply ; diagnostic imaging ; metabolism ; pathology ; physiopathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Perfusion Imaging ; Pulmonary Circulation ; Receptor, EphB4 ; metabolism ; Tomography, Spiral Computed ; methods

To construct plasmid of recombinant protein candidate vaccine of respiratory syncytial virus, express it in E. coli, and to investigate its immunogenicity and protective efficacy. A CD8+ T cell epitope from respiratory syncytial virus (RSV) M2 protein F/M2:81 - 95 and the G:125-225 (G1) gene fragments from RSV-G protein containing B cell epitopes were amplified by PCR method and then inserted into the prokaryotic expression vector pET-DsbA after bonding to a linker. The fusion protein DsbA-G1-Linker-F/M2:81-95 (D-G1LF/M2) was expressed successfully in E. coli BL21 (DE3). The product was proved to be RSV-specific by Western-blot. After purified by affinity chromatography on Ni+ Sepharose and renatured by gradient dialysis. D-G1LF/M2 was used to immune BALB/c mice. D-G1LF/M2 induced high anti-D-G1LF/M2 IgG, anti-RSV IgG and neutralizing antibody titers in serum and lung of BALB/c mice, and elicied RSV-specific CTL responses. The IgG subclass distribution revealed that IgG1/IgG2a ratio was 2.66. Viral titration indicated that D-G1LF/M2 could protect BALB/c mice against RSV challenge in lung.
Animals ; Antibodies, Viral ; blood ; immunology ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin G ; blood ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Respiratory Syncytial Virus Infections ; prevention & control ; Respiratory Syncytial Virus Vaccines ; biosynthesis ; genetics ; immunology ; Respiratory Syncytial Virus, Human ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; Viral Fusion Proteins ; genetics ; Viral Proteins ; genetics