Objective Changes of thyroid stimulating antibody(TSAb) and thyroid stimulating blocking antibody(TSBAb) in the treatment of anti-thyroid drugs(ATDs), and the effect of ATDs combining with levothyrocine(LT4) on TSAb and TSBAb were analyzed. Methods Using recombinant Trxfus. TSHRn protein and Trxfus. TSHRc protein as antigens, and TSH receptor antibody(TRAb)-N(TSAb binding hot spots), TRAb-C(TSBAb binding hot spots)in the serum of thyroid disease patients were measured with ELISA. The changes of TRAb-N, TRAb-C over 36 months in 117 TRAb-N positive Graves′ patients with hyperthyroidism were analyzed retrospectively. In the course of treatment, 41 cases as A group with ATDs and LT4 treatment, 76 cases as B group with only ATDs, The changes of TRAb-N and TRAb-C were observed in the two groups. Results (1)According to the change of TRAb-N, 117 TRAb-N positive Graves′ patients with hyperthyroidism were different. In group Ⅰ, 10 patients continued to have persistently positive TSAb and continued to have hyperthyroidism, remission rate 0%. In group Ⅱ, 17 patients showed complicated TRAb-N changes, 12 of 17 patients got relapse, 5 of 17 patients got remission, remission rate 29.4%. And in group Ⅲ, with TRAb-N dropping gradually, 15 of 89 patients got relapse, 74 of the 89 patients got remission, remission rate 83.1%. Three groups were significantly different with x2 test(P<0.01). One of the 117 TRAb-N positive Graves′ patients with hyperthyroidism developed TRAb-C positive hypothyroidism. (2)According to combining with and without LT4 during the treatment of ATDs,the patients were divided into 2 groups(Group A: ATDs combined with LT4; Group B: only ATDs). These 2 groups were significantly different in TRAb-N at baseline and 3 months(P<0.01), TRAb-C between two groups were not significantly different in all times(P>0.05). Conclusion TSAb and TSBAb can be used to document TRAb-function, which is significant for us to predict the changes of thyroid function. During ATDs treatment, the temporary early low-dose application of LT4 did not significantly affect TSAb and TSBAb.

Objective To select and express a human thyrotrophin receptor antibody (TRAb) Fab fragment from phage antibody library constructed with phage display technology. Methods With immobilized antigen, the reconstructed humanized TRAb Fab library was enriched by five rounds panning (adsorption-elution-amplification). The TSAb Fab and TBAb Fab fragment were selected by coated fusion proteins of hTSHRn and hTSHRc. The positive clones were identified and selected by Phage-ELISA. TRAb positive clones were identified by PCR and double restriction enzyme digestion. The soluble TRAb (TSAb, TBAb) Fab fragments were expressed. TRAb (TSAb, TBAb) Fab fragments were identified by Western blotting assay. The DNA fragment was sequenced from the positive clones. Results Following five rounds of biopanning, TRAb (TSAb,TBAb) Fab phage antibody was screened. The enrichment effect reached to 77 times and 94 times. The soluble TRAb (TSAb,TBAb) Fab antibodies were expressed from positive clones and identified by phage ELISA. Western blotting analysis showed that the phage displaying Fab had significant binding activity with antigens. These sequence analysis showed that all of the heavy chain Fd gene and light chain gene were derived from human immunoglobulin variable region. The light chain variable region of the monoclonal 48 was homologous to the immunoglobulin light chain Vλ homology of 94.4%, and the heavy chain variable region of the monoclonal 48 was homologous to the immunoglobulin heavy Fd chain VH4 homology of 88.9%. The light chain variable region of the monoclonal 56 was homologous to the immunoglobulin light chain Vλ homology of 95.6%, and the heavy chain variable region of the monoclonal 56 was homologous to the immunoglobulin heavy Fd chain VH3 homology of 84.6%. Conclusion We have successfully selected TRAb (TSAb, TBAb) Fab fragment from a human phage display immune antibody library.

Objective To observe the effect of Cyclosporine A on the brain immunoligical reject reaction of transgenic cellular transplant in rats with experimental brain hemorrhage.Methods We established the brain hemorrhagic models of rats and the models were divided into group A (treated with Cyclosporin A) and group B (without treatment) randomly after transplanting brain cells modified by NGF gene. The levels of CD~+_4, CD~+_8T cells subsets in peripheral blood were examined by using flow cytometry at 15 d after transplantation. The expressions of MHC-classⅡantigens and infiltration of CD~+_4, CD~+_8T lymphocyte subsets were examined by SP immunocytochemical stain.Results The count of CD~+_4, CD~+_8T lymphocyte and CD~+_4/CD~+_8 in peripheral blood were 29.20?3.97, 20.65?2.02 and 1.41?0.86 in group A, and were 47.39?3.01, 28.30? 2.36 and 1.68?0.69 in group B, respectively.There were significant differences between the two groups (all P

BACKGROUND: A stable and exact animal model is the necessary tool and basis for studying hemorrhagic cerebrovascular diseases.OBJECTIVE: To establish and evaluate the intracerebral capsular hemorrhage models in rats.DESIGN: A randomized and controlled animal experiment.SETTING: Second Hospital Affiliated to Xuzhou Medical College; First Hospital Affiliated to Nanjing Medical University.MATERIALS: This experiment was carried out in the animal experimental center of Nanjing Medical University during May to November 2002.Totally 35 SD rats were randomized into two groups: experimental group (n=30) and sham-operation group (n=5).METHODS: ① Autoblood was injected into the intracerebral capsule of rats to create intracerebral capsule hemorrhage models with stereotaxy in the experiment group. ②Scoring was conducted according to 5-point neurological scoring criteria from ZeaLonga, somatic sensation and motor function of rats were observed. ③Somatosensory evoked potential(SEP) of rats was detected pre- and post-operation under anesthetic state. ④ After determination of SEP, the rats were sacrificed under anesthetic state. Brains were taken out to made slices, then sections were stained with haematoxylin and eosin. Changes in haematoma and histomorphology were observed at the largest focus under optical microscope.MAIN OUTCOME MEASURES: ①Nerve function scoring; ②Latent period of various waves of SEP; ③ Observation of brain tissue morphology.RESULTS: Totally 35 rats entered the stage of result analysis. ①Appearance of obvious paralysis of the rats suggested the modeling was successful. The successful rate of this experiment was 93.3%(28/30). Significant difference existed in neurological scoring between experimental group [(2.74±0.46)points] and sham-operation group (0 point)(P＜0.05). ②SEP showed that the latent periods of various waves of experimental group after operation were significantly delayed than those before operation and those of sham-operation group [P1: (15.72±0.78) ms, (10.69±0.52) ms, (10.73±0.48) ms;Nl: (17.95±1.27) ms,(13.21±1.31) ms, (13.34±1.27) ms;N2:(21.16±1.62) ms, (15.42±1.46) ms,(15.58±1.44) ms;N3:(24.86±1.58) ms, (18.72±1.76) ms, (18.99±1.67) ms,P＜0.05]. ③In the shamoperation group, a few red blood cells were scattered in the peripheral area of needle channel were found, but hemorrhagic focus was not; In the experimental group, irregular or oval blood clots presented in the left internal capsule area. In about a low-fold visual field, brain tissue in the surrounding of hemorrhagic focus was loosened and swelled, and pathological changes were obviously severer than those in the sham-operation group.CONCLUSION: Intracerebral capsular hemorrhage induced by injection of autoblood with stereotaxy is more close to clinical situation, and it is easy to operate and has good reproducibility.

Objective To investigate the physicochemical characteristics of urinary stone induced by ceftriaxone.Methods Two children cefriaxone-associated urinary stone samples were received for component analysis in our hospital in April 2012,of which one was from a boy whose clinical data was not available,and the other was from a boy who suffered acute lower abdominal pain and vomiting after treatment with ceftriaxone for 5 d in early April.Ultrasound demonstrated a stone in his right upper ureter.Computed tomography showed right upper ureteral stone,which was radiolucent on plain abdominal radiograph.After a conservative treatment for 3 d,the stone in right upper ureter was spontaneously passed,which was confirmed by the ultrasound and intravenous pyelography.The two received stone samples were analyzed by infrared spectroscopy,scanning electron microscopy and energy disperse spectroscopy for component analysis.Results The compositions of two stone samples were free ceftriaxone and calcium ions combined with a molar ratio of 1:1.Conclusions The ceftriaxone could induce urinary stone in children.This special stone has radiolueent imaging,and it is composed of calcium ceftriaxone salt.

ObjectiveTo observe the changes in TSH receptor antibody (TRAb) and soluble intercellular adhesion molecule-1 ( sICAM-1 ) levels after treatment of propylthiouracil ( PTU ) and methimazole( MMI ) in patients with Graves' disease (GD). MethodsOne hundred and six cases of clinically diagnosed patients with GD were divided into PTU and MMI groups( each group, n =53 ). The patients in two groups were regularly followed for 36 months. TRAb and sICAM-1 were measured with ELISA method. Results( 1 ) The general data of two groups were not significantly different before treatment( all P＞0. 05 ). ( 2 ) There was no difference in TRAb positive rates between two groups before treatment. The clinical remission rates between PTU and MMI groups showed no difference (83.02％ vs88.68％ ). The cure rate was higher in MMI group than in PTU group( 58.49％ vs 37.74％, P＜ 0. 05 ). (3) There existed significant differences in TRAb levels determined before and 6, 12, 24,30, and 36 months after treatment( all P＜0. 01 ), being gradually decreased with time( F=275.48 ,P＜0.01 ). TRAb levels between two groups were significantly different( F=5.86, P＜0. 05 ). (4) sICAM-1 levels at 36 months after treatment compared with the baseline in both groups were statistically different (P＜0. 01 ), but no difference was found between two groups.ConclusionsBoth PTU and MMI improve the immune status of patients with GD,and the immunosuppressive effect of methimazole is more evident.

Objective The objective of this study was to observe the effect of AEG-1 gene on the metastasis in breast cancer MCF-7 cells.Methods AEG-1 siRNAs were transfected into MCF-7 cells to silence AEG-1 expression,and negative siRNA was used as a control.Transwell chamber was used to detect the ability of cell migration and invasion of MCF-7 cells.CCK8 assay was used to detect the cell proliferation of MCF-7 cells.At the same time,the effect of VEGF on the angiogenesis was investigated by detecting the changes of lumen formation in HUVEC cells.Results The migration,invasive and proliferative abilities were significantly inhibited in MCF-7 cells transfected with AEG-1 siRNA.Knockdown AEG-1 was significantly decreased the level of VEGF in the supernatant of MCF-7 cells.Knockdown AEG-1 was also significantly inhibited the angiogenesis activity in HUVEC cells.Conclusion Knockdown AEG-1 can significantly inhibit the migration of MCF-7 cells,including cell migration,invasion,proliferation and angiogenesis.These results suggest that AEG-1 plays an important role in the metastasis process of breast cancer and opens up new ideas for future treatment breast cancer.

BACKGROUND:The metastatic potential of hepatocelular carcinoma cels is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cels on metastasis of hepatocelular carcinoma is of great significance for improving the lifetime of hepatocelular carcinoma patients. OBJECTIVE:To explore the biological effect of human bone marrow mesenchymal stem cels on hepatocelular carcinoma cels with different metastatic potentials. METHODS:Human bone marrow mesenchymal stem cels and hepatocelular carcinoma cel suspension with high and low metastatic potentials were respectively injected into the Transwel chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as wel as cel counting method was used to observe the changes in the invasion ability of hepatocelular carcinoma cels. The effects of human bone marrow mesenchymal stem cels on the proliferation of hepatocelular carcinoma cel suspension with high and low metastatic potentials were determined using cel counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cel death protein 5. RESULTS AND CONCLUSION:(1) The number of migrated hepatocelular carcinoma cels was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P> 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group (P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P< 0.05), but the expression of programmed cel death protein 5 showed no changes (P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cel death protein 5 was both increased dramaticaly (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cels reduce the invasion ability of hepatocelular carcinoma cels, and enhance their ability of proliferation.

Objective:To explore the optimal postpartum nursing intervention method of elderly second-birth parturient based on the best evidence and evaluate its effect so as to improve the quality of clinical nursing.Methods:Based on the best evidence obtained from the previous study, the optimal nursing examination index of the elderly second parturient was established, and the evidence of best practice was obtained by using the evidence-based nursing method, and the 2017 revised version (the third edition of) Johns Hopkins evidence-based nursing practice model) was used to guide nurses to apply the evidence to clinical practice. During the period from March to May 2019, the baseline review was completed, including 82 pregnant women and 43 nurses. The evidence was incorporated into clinical practice in June 2019. From July to October 2019, the same indicators and inclusion and exclusion criteria were used to evaluate the effectiveness of evidence application in 43 nurses (including midwives) and 85 elderly second-child parturients in obstetric wards (including baby-friendly areas). The anxiety, depression, maternal satisfaction and nurses' correct implementation rate of the best evidence were compared before and after the application of evidence.Results:After the use of evidence-based intervention, the implementation rate of nurses was above 80%. The anxiety score of parturient decreased from 55.29±10.11 to 41.74±10.35, the difference was statistically significant ( t value was 8.55, P<0.001), and the depression score decreased from 57.77 ±8.32 to 47.86 ±9.33, the difference was statistically significant ( t value was 7.23, P<0.001). Maternal satisfaction increased from 92.68% (76/82) to 98.82% (84/85), the difference was statistically significant ( χ2 value was 3.92, P< 0.05). Conclusion:The result of practice shows that through evidence-based nursing intervention, we can further standardize the postpartum optimal nursing paradigm of elderly second-child parturients in China. Reduce postpartum anxiety and depression and improve maternal satisfaction. And quality review still needs to be continuously promoted and implemented in clinical practice in order to continuously improve the quality of nursing and ensure sustainability.

Objective To assess the distribution of vaginal microflora in healthy women and investigate their cognition of lower reproductive tract infection-related knowledge as well as personal hygiene habits and character of behavior in seeking medical treatment. Methods Total of 1660 healthy women who had physical check-ups at the Medical Center of Renji Hospital were selected and received gynecologic examination, as well as routine examination, pH examination and bacterial culture of the vaginal discharge. In addition, 860 of them were randomly selected for questionnaire survey, in which reproductive tract infection -related knowledge and personal hygiene habits as well as behavior in seeking medical treatment were involved. Results Among the 1660 eensused women, vaginal average pH was 4. 16±0.21. The positive rate of Candida in vaginal discharge routine examination was 3.86% (64/1660), which was lower than that in aerobes culture 7.71% (128/1660). Candida alhicans was the most populous species 78.9% (101/128) ,followed by 7.8% (10/128) and 7.0% (9/128) for the Candida glabrata and Candida krusei respectively. The most populous species of vaginal microflora were hemolytic streptococcus A (63.80%, 1059/1660), Staphylococcus epidermidis (14.28%, 237/1660), and Enterococcus faecalis (D) group ( 11.44%, 190/1660). The questionnaire survey showed that 88.4% (760/860) of 860 women took active treatment when feeling unwell, 92.1% (792/860) of them had good hygiene practices, and only 21.2% (182/860) had the habit of vaginal douching. In addition, 50.0% (430/860) of them had the desire to obtain reproductive health knowledge through out-patient consultation. Conclusions Vulvovaginal Candida disease ranks the first in all types of vaginitis, among which, Candida albicans is the most populous species followed by the Candida glabrata and Candida krusei. Hemolytic streptococcus A and Staphylococcus epidermidis are the most common species of vaginal mieroflora in healthy women. The censused women have high awareness of reproductive health care and pay mueh attention to common gynecologic diseases such as lower reproductive tract infection. Nevertheless, we should strengthen the public education of reproductive health-related knowledge.