Objective:To establish and evaluate a newly established method of enzyme-linked immu-nosorbent assay (ELISA) for measuring human autoantibody to folate receptor (FR).Methods: Folate receptor was extracted and purified from healthy woman placenta tissues .The protein was coated on 96-well plates.Goat monoclonal antibody was used as detecting antibody to set up the indirect ELISA proce -dure.The sensitivity, precision and linearity of the method were evaluated .Further, the method was compared with the ELISA method with commercialized bovine folate binding protein ( FBP) by determi-ning autoantibody levels in 24 individuals .Results:The measuring range of the standard curve was from 6 .25 ×10 -4 to 8 ×10 -2 ( the IgG concentration of pooled plasma from healthy donors was defined as 1 ) . The lowest detectable level was 3.13 ×10 -4 .The intra-and inter-assay coefficients of variations were 2.74%-8.07% and 4.16% -8.23%, respectively.Linearity test results were considered within acceptable limits.The data from FBP-ELISA and FR-ELISA were highly correlated ( r=0.954, P <0.001);The value from FR-ELISA was higher by 14% than that from FBP-ELISA.Conclusion: The ELISA method for measuring human autoantibody IgG to folate receptor was successfully established using human FR as coating protein .The method is sensitive and repeatable and can be used in large-scale population study .

Objective To explore the clinical value of expression of CEA and CK-20 detected by flow cytometry (FCM) in peritoneal washes on predicting peritoneal metastasis and prognosis of gastric carcinama.Methods Clinicopathological characteristics and follow up data of 105 patients with gastric carcinoma who were underwent D2 radical resection (R0) were collected.Peritoneal washes was collected.Peritoneal lavage cytology examination (PLC) was used to find intraperitoneal free cancer cells (IFCC).FCM was used to determine the CEA and CK-20 expressions.Meanwhile,13 patients with benign lesion on the stomach and the gastric carcinama cell line SGC-7901 were served as the negative and positive control,respectively.Results Positive expression of CEA was in 48 (45.7 ％) patients,of CK-20 was found in 67 (63.8 ％) patients,and of CEA and CK-20 was in 85 (81.0 ％) patients by FCM.However,positive expression of IFCCs was found in 31 patients(29.5 ％) by PLC.The expression of CEA and CK-20 were related to serosa invasion,lymphnode metastasis and pTNM stage (P ＜ 0.05).The median survival of patients with negative expression of both CEA and CK-20 (n =20) was significantly longer those with positive expression (n =30)(52 vs 18 months,P ＜ 0.05).Conclusions Combined Detection of CEA and CK-20 in peritoneal washes by FCM can be used to predict peritoneal micrometastasis and may predict the prognosis of gastric carcinoma.

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.

Objective To investigate the effects of hypoxia on the expression of Stat3 and p-Stat3 ,and assessed the apoptosis ability of JAR cells in vitro .Methods JAR cells were cultured under hypoxic conditions .Western blot were used to determine the protein expression of Stat3 and p-Stat3 .Cellular apoptosis was monitored by flow cytometry analysis .Results Abnormal morpholo-gy changes in trophoblast cells under low oxygen conditions .After 48 h hypoxic treatment ,the protein of Stat3 and p-Stat3 were significantly decreased(P< 0 .05) ;however ,the level of apoptosis was significantly increased (P < 0 .05) .Conclusion Stat3 and p-Stat3 protein levels were decreased under hypoxia circumstance ,while the cell apoptosis ability was increased in JAR cells .

Objective To study the effects of miR?21 on the proliferation,apoptosis and MMP2 expression of HeLa cells. Methods The miR?21 or the miRNA scramble control was transfected into Hela cells. The cell proliferation was detected by the Celltiter?GloTM assay 72 h after the transfection,and the apoptosis was evaluated by the Caspase3/7 Glo? Kit. The MMP2 RNA expression was quantified by quantitative real time PCR. Results The proliferation of HeLa cells transfected with miR?21 was significantly increased compared to that of the cells transfected with miRNA scramble control. The caspase 3/7 activity in HeLa cells transfected with miR?21 was downregulated compared to that of the cells transfect?ed with miRNA negative control. The MMP2 RNA expression in HeLa cells transfected with miR?21 was increased significantly compared to the cells transfected with miRNA negative control. Conclusion miR?21 can significantly promote the proliferation of HeLa cells. In addition,it ex?hibits the anti?apoptosis effect in HeLa cells. The transfection of miR?21 significantly increased the MMP2 RNA expression,which suggests that miR?21 may promote the tumor invasion and might be a therapeutic intervention target.

Objective To analyze the clinical characteristics and prognosis of disseminated cryptococcosis in children.Methods The data of disseminated cryptococcosis inpatients were reviewed retrospectively at Beijing Children's Hospital,Capital Medical University,from January 2002 to September 2014.The demographic data,clinical manifestations,laboratory findings,imaging,antifungal treatments and outcomes of all the patients were analyzed.Results Overall 25 children with disseminated cryptococcosis were enrolled including 17 boys(68.0％).The average age was 7 years old.Four cases (16.0％) had underlying conditions,among them 1 case had human immunodeficiency virus(HIV)-positive.The median time to diagnosis was 32 (23-47) days,44.0％ of the patients (11 cases) were misdiagnosed,and 8 cases misdiagnosed as tuberculosis.All patients had fever.Other common clinical manifestations included cough (16 cases),headache (10 cases),vomiting (10 cases),altered mental status (6 cases) and stomachache (6 cases).Respiratory system involvement was seen in all cases,central nervous system was involved in 18 cases,other organ involvement included lymph nodes,spleen,liver,kidney,skin,skeleton and costicartilage.Amphotericin B (AmB) + Fluconazole (Flu) ± 5-flucytosine (5-FC) was the most common therapy (15 cases),Flu ± 5-FC for 6 cases,AmB ± 5-FC for 2 cases,Voricanazole (VOR) for 1 case.Sixteen cases (66.7％) got recovery/improved on discharge,8 cases (33.3％) rejected to the advise and discharged with treatment failure,and the HIV-infected patient transferred to a special hospital.Fifteen patients (60.0％) were followed up,and 13 cases (87.0％) showed recovery,but 2 died in the long-term prognosis.Conclusions Disseminated cryptococcosis in children is more common in school-age boys.Most patients are without underlying conditions.Disseminated cryptococcosis can cause multiple organ damage.Patients with prolonged fever,cough,headache,with or without underlying diseases,can be suspected as disseminated cryptococcosis.Blood and cerebrospinal fluid culture for fungus,cryptococcal antigen should be performed for early diagnosis and treatment.

Objective To Compare characteristics between the psychological aids hotline callers who had single attempted suicide and those callers who had repeated attempted suicide. Methods The analysis was conducted on the da?tabase of Beijing psychological aids hotline from Dec., 2002 to Dec., 2008. All the effective callers were consulted by pro?fessional hotline operators. During the calling, the callers were interviewed on their attempted suicides, depression and 11 correlates associated with caller's suicidal behaviors. Results Among 4519 callers who had attempted suicide, 2441 (54%) of them had attempted suicide once, and the other 2078 (46.0%) callers had repeated suicide attempts. After ad?justed for gender, age, and the other 4 demographic variables, history of being abused (OR=1.35, 95%CI: 1.08~1.67), afraid of being attack (OR=1.35, 95%CI: 1.01~1.59), relatives or associates with prior suicidal behavior history (OR=1.17, 95%CI: 1.001~1.36), hopelessness (OR=1.20, 95%CI: 1.02~1.40), and psychological treatment history (OR=0.73, 95%CI:0.62~0.86) were associated with repeated suicide among hotline callers. Conclusions To improve the effective?ness of hotline based suicide prevention, assessment of risk of repeated attempted suicide among the callers with a histo?ry of prior attempted suicide should focus on the history of being abused, afraid of being attack, hopelessness, relatives or associates with prior suicide history, and psychological treatment history.

Objective To observe the effect of wheel running and treadmill running on the cognitive functioning of rats after the extraction of a molar.Methods Thirty-two Sprague-Dawley rats were randomly divided into a control group (group C,n =8),a molar extraction group (ML group,n =8),a wheel running pretreatment group (W-ML group,n=8) and a treadmill running pretreatment group (T-ML group,n =8).The rats in the T-ML and W-ML groups practiced wheel running and treadmill running for 1 week,then they and the rats in the ML group received molar extraction surgery.One week later,a passive avoidance test was used to evaluate the cognitive functioning of all of the rats.Results No significant differences were found in the groups' average latency I times,but the average latency II time of the ML group was significantly shorter.There was no significant difference in the average latency II times of the T-ML and W-ML groups.Conclusion Either wheel running or treadmill running can promote learning and memory after molar loss by oral surgery,at least in rats.

Objective To explore the regulatory effects of HTLV-1 ( human T-cell leukemia virus type 1 ) Tax protein on the expression of HMGB 1 ( high mobility group box 1 ) gene in T cells .Methods Total RNA and protein were extracted from Tax +-T cells ( TaxP ) , Tax--T cells ( TaxN ) and Jurkat cells which were stably transfected with pCMV-Tax and pCMV-Neo, respectively.Then, the expression levels of HMGB1 mRNA and protein in different CD 4+T cells were analyzed by real-time PCR and Western blot (WB).By using liposome-mediated method, pGL3-HMGB1-luc reporter genes and pGL3-neo-luc were tran-siently transfected into TaxP and TaxN cells and the basal transcriptional activity was observed in different T cells.Additionally, pCMV-Tax and pGL3-HMGB1-luc reporter genes were also co-transfected into Jurkat cells and the regulatory effects of Tax protein on HMGB 1 gene was detected .The chromatin immunoprecipi-tation (ChIP) assay was used to identify HMGB1 genomic sites directly targeted by Tax .Results The ex-pression levels of HMGB1 mRNA and protein in Tax+-T cells ( TaxP) were higher than those in Tax--T cells (TaxN).The transcription regulation trends for HMGB1 gene in TaxN and TaxP cells were similar but not identical in diverse T cells.pHLuc3 (containing -504-+83 HMGB1) showed the highest transcriptional ac-tivity of HMGB1 gene in both TaxP and TaxN cells , but HMGB1 transcriptional activity of pHLuc 6 in TaxP cells was significantly stronger than that in TaxN cells .Luciferase assays also showed that Tax protein promo-ted the transcription of HMGB1 gene in a dose-dependent manner .The ChIP assay further confirmed that Tax protein enriched at the HMGB1 region of -1163--1043.Conclusion The region of nt -504--383 is essen-tial for the basal promoter activity of -1163-+83 HMGB1 gene originated from pHLuc 6 reporter plasmid , and Tax protein enriched probably at the HMGB 1 site of -1163--1043 enhances HMGB1 transcription.

Objective:To investigate the optimal monoenergetic level of spectral reconstructions and contrast agent concentration for the structure and lumen of isolated coronary stents on a dual-layer spectral detector CT (DLCT).Methods:Eighteen scaffolds of different materials were implanted into the artificial simulated coronary artery, and were filled with conventional and low concentration contrast agents (1/2 of the conventional dose). Spectral CT scan was performed, and 50-100 keV (interval 10 keV) and 120 keV single-level reconstruction were performed at the conventional concentration and low concentration, which were recorded as A 50-100, 120 keV group and B 50-100, 120 keV group respectively; Another group of conventional concentration data was reconstructed with conventional iDose 4, which was recorded as conv group. The image was evaluated subjectively, including stent structure score (Likert 5-score) and stent lumen score (Likert 4-score); objective evaluation, including background noise, contrast noise ratio (CNR) and stent inner diameter difference (AlN). The differences of subjective and objective evaluation indexes were analyzed. Results:The stent structure scores of A 80, 90 keV group [(3.56±0.64), (4.11±0.58) points] and B 90 keV group [(3.61±0.50) points] were significantly better than those of other groups ( P<0.01), and the A 90 keV group had the highest score. The scores of stent lumen in A 70 keV group [(3.61±0.50) points] and B 70 keV group [(3.47±0.63) points] were significantly better than other groups ( P<0.01), and the score of A 70 keV group was the highest. There was no significant difference in noise, CNR and AlN ( F=1.67, 0.15, 1.16, P>0.05). The main effect analysis was energy level, there were significant differences in noise, CNR and AlN between groups a, B and conv ( F=83.74, 23.95, 24.63, P<0.01). A 70, 80, 90 keV group [(11.43±0.48), (11.81±0.34), (12.01±0.33) HU] and B 70, 80, 90 keV group [(11.50±0.31), (11.58±0.18), (12.13±0.30) HU], image background noise was significantly lower than other groups ( P<0.01). There was no significant difference between the six groups ( P>0.05). The noise of A 70 keV group was the least. The CNR in stent lumen of A 50, 60, 70 keV groups (26.85±5.19, 26.03±4.29, 24.46±5.40) and B 50, 60, 70 keV groups (25.88±5.67, 24.18±5.46, 24.91±7.67) was significantly higher than that of other groups ( P<0.01). The highest CNR value was found in A 50 keV group. The AlN value of A 90, 100, 120 keVgroup [(32.22±6.16) %, (30.79±5.55) %, (27.05±4.47)%] was higher than that of conv group [(41.76±6.00)%], the difference was statistically significant ( P<0.01). The AlN values of A 90, 100, 120 keV group and B 90, 100, 120 keV group [(34.34±5.73)%, (33.95±5.46)%, (33.92±5.45)%] were lower than those of other subgroups, and the differences were statistically significant ( P<0.01). There was no statistically significant difference between the six groups ( P>0.05). The AlN of A 120 keV group was the lowest. Conclusions:Regarding stent phantom imaging, DLCT monoenergetic imaging was better than conventional mixed energy imaging. When DLCT scanning was performed, 90 keV (structure) and 70 keV (lumen) combined reconstruction could not reduce the imaging quality when the concentration was halved.