Objective To screen the tumor specific antigens, and explore new methods for diagnosis and therapy of tumor. Methods Hela cells, the human cervical carcinoma cell line, were used to immunize the BALB/C mice, and the positive hybridomas were screened by rosette formation screening system in human carcinoma cell line Colo205, Burkitt lymphoma cell line, Raji, etc. The immunofluorescence staining and flow cytometry analysis were performed to characterize if the prepared monoclonal antibodies (mAb) could reorganize the natural membrane antigen expressed on Hela, Colo205, Raji, PLA801, 7901, Jurkat, 293T, ECV304 and resting or activated human peripheral blood mononuclear cells (PBMC). Furthermore, the membrane protein of Hela, Raji and Sp2/0 cells was segregated by ultracentrifugation and the mAbs were cross-linked with the Sepharose CL-4B. In order to identify the molecules which could be recognized by the prepared mAbs, immunoprecipitation was performed to purify the tumor antigen candidates with the cell membrane protein from Hela, Raji and SP2/0 (as negative control) through Sepharose CL-4B cross-linked with the prepared mAbs. Finally, western blotting was performed to detect the molecular weight of the products from the immunoprecipitation. Results Three mAbs were obtained which could recognize the tumor antigen candidates expressed in several human carcinoma cell lines, such as Hela, Raji and Colo205, and some embryonic cell lines, such as 293T and ECV304, but they could not bind to Jurkat, PLA801, 7901, SP2/0, resting or activated human PBMC. Western blotting results indicated that the three mAbs could recognize a 47kD molecule which expressed in Hela and Raji but not on SP2/0 cell membrane. Conclusion Three mAbs, which can recognize tumor membrane antigen candidates, are obtained, providing useful tools for screening and identify the tumor specific antigens.

Objective To detect the expression of scleroderma-related autoantibodies, such as anti-Scl-70, anli-centromere antibody ( ACA)and anti-RNA polymerase Ⅲ ( ARA) , and their relationship with clinical features in Chinese systemic sclerosis (SSc) patients. Methods One hundred and thirty-five Chinese SSc patients from the clinical database of the Scleroderma Trials and Research Group proposed by European League Against Rheumatism's Scheroderma Trial and Research Group( EUSTAR) were consecutively enrolled. The expression of ARA, anti-Scl-70 and ACA were detected through linear immunoblotting, double immunodiffusion and indirect irnmunofluorescence, respectively. The relevance between the existing of autoantibodies and clinical manifestations was analyzed statistically. Results Among the 135 Chinese SSc patients, the prevalence of anti-Scl-70, ACA, ARA were 49. 6% , 13.3 % and 8.9% respectively. Patients with anti-Scl-70 antibody had significantly shorter disease course [(71 ±59) month vs (90 ± 103) month, P = 0.041] , higher proportion of interstitial lung disease ( P = 0. 031) but lower of pulmonary arterial hypertension (P =0.042). Modified Rodnan's skin score (P=0.008) and prevalence of facial and cervical cutaneous sclerosis (P = 0. 002) , distal (to elbow/knee ) cutaneous sclerosis ( P = 0. 004 ) and digital pitting scarring/disappear of digital pad were all significantly higher in anti-Scl-70 positive group. Patients with AC A had longer disease course ( P = 0. 036) , lower IgM level ( P = 0. 045) and were less prevalent of interstitial lung disease ( P =0. 045). Patients with ARA had higher serum creatinine and urea nitrogen level ( P ＜ 0.001) although otherwise features had unremarkable differences. Conclusion Scleroderma-related autoantibodies have relevance with different clinical manifestation and detection of these autoantibodies may be helpful to the diagnosis of SSc, organ involvement evaluation and predicting outcomes. The clinical relevances of autoantibodies in Chinese SSc patients may differ from other areas or races.

Objective To investigate the biological functions of IncRNA-BG on the radiosensitivity of normal human bronchial epithelial cell line Beas-2B.Methods Three IncRNA-BG siRNAs were designed,synthesized and traasfected into Beas-2B cells via lipofectamine.The RNA transcription level of BG was detected by quantitative real time-PCR to confirm the siRNA transfection efficiency.The experiment was divided into control group,control siRNA transfected group,and BG transfected group.Cell survival was detected by clonogenic assay,and the cell cycle distribution was determined by flow cytometry assay.The γ-H2AX foci formation after irradiation was visualized via immunofluorescence.Western blot assay was performed to detect the protein expressions of RAD50,p-P53,KU70,KU80,MDM2,CDK2 and RB.Results BG-siRNA transfection significantly reduced the BG transcription level (t =8.32-15.29,P ＜0.05) and increased cell survival after irradiation at 0.5,1,2,4 and 6 Gy.Analyzed with the multi-target model,the SERD0 of Beas-2B cells and control siRNA transfected cells were calculated to be 0.80 and 0.82,respectively.In addition,BG-siRNA transfection enhanced radiation-induced cell cycle arrest at G2 phase so that,after 4 Gy irradiation,the cells in G2 phase was increased from (37.37 ±0.63) ％ of control siRNA cells to (64.19 ± 1.01) ％ (t =30.65,P ＜ 0.05).Meanwhile,the γ-H2AX foci of BG-siRNA transfected cells was decreased from 76 ± 1.78 per 100 cells to 59-± 3.49 per 100 cells (t =13.72,P ＜0.05).The expressions of DNA damage related proteins including KU70,KUS0,CDK2 and RB were increased,but the expressions of p-P53 and RAD50 were decreased.Conclusions LncRNA-BG could regulate the radiosensitivity of the normal human bronchial epithelial cells,probably through inducing cell cycle G2 phase arrest and promoting DNA damage repair after irradiation.

Objective To observe the effect of Shengji corium elephatis mastic in treating chronic refractory skin ulcer, and analyze its preliminary mechanism. Methods Totally 62 patients with chronic refractory skin ulcer in granulation stage were randomly divided into two groups, 32 cases of treatment group were treated with Shengji corium elephatis mastic, and 30 cases of control group were treated with Vaseline gauze. All patients were treated for 4 weeks. The rate of wound healing, wound reduction ratio and wound secretion level of VEGF were observed. Results The cure rate and the total effective rate between the two groups had significant difference (P<0.05). After 2 weeks treatment, the mean wound reduction area of treatment group and control group was 82.31%and 66.32%respectively. After 4 weeks treatment, the mean wound reduction area of treatment group and control group was 90.35%and 78.7%respectively, the difference was statistically significant (P<0.05). After 1, 2 and 3 weeks treatment, the treatment group had significant difference with the control group in wound secretion VEGF level (P<0.05). Conclusion Shengji corium elephatis mastic can promote wound healing of chronic refractory skin ulcer. The possible mechanism is that Shengji corium elephatis mastic promotes the generation of VEGF in wound thus promotes wound repair.

Objective To identify a novel auto-antibody in sera of systemic sclerosis (SSc) patients and to analyze its relevance with SSc-associated interstitial lung disease (ILD). Methods The anti-moesin antibody in the sera of 62 SSc patients, who had participated the European League Against Rheumatism's Scl eroderma Trial and Research Group (EUSTAR), were tested by enzyme linked immunosorbent assay (ELJSA). Patients were grouped by high resolution computerized tomography (HRCT) features, pulmonary function test (PFT) abnormalities, inflammatory markers and disease course. The prevalence and titer (Optical density value) of anti-moesin antibody were compared between groups with t and χ2 test. Results The titer of anti-moesin antibody was significantly higher in the SSc-ILD group than non-ILD group (0.156±0.062 vs 0.107± 0.026, P=0.005). Among SSc patients, the diagnostic sensitivity and specificity of the anti-moesin antibody for ILD was 44.0% and 91.7% respectively (Kappa=0.2, P=0.022). Anti-rnoesin antibody was more prevalent in SSc patients with HRCT features of honeycomb-like lesion, lobular septal thickening and mediastinal lymphadenopathy (P<0.05). SSc patients with deteriorated total lung volume (TLC %) had higher titer of anti-moesin antibody significantly (0.172±0.067 vs 0.133±0.039, P=0.011), as the same tendency in patients with decreased diffusing capacity of the lung for carbon monoxide (DLco% ) but without statistical significant difference (0.153±0.580 vs 0.120±0.340, P=0.089). The anti-moesin antibody was equally prevalent between abnormal ESR, C reactive protein, immunoglobulin and complements groups and their normal controls (P> 0.05). Group of patients who had SSc courses more than or less than 5 years demonstrated similar anti-moesin antibody titers (0.146±0.047 vs 0.164±0.077, P=0.272). However, patients with ILD courses less than 12 months had higher liter of the antibody than controls (0.182±0.073 vs 0.138±0.049, P=0.040). Conclusion This study suggests that the novel anti-moesin antibody has comparatively high specificity for SSc-associated ILD patients, which may contribute to further understanding the pathogenesis of ILD in SSc patients. Further investigations are deserved to evaluate the application of anti-moesin antibody in facilitating early screening and evaluation of ILD.

Objective To observe the therapeutic effect of hoop circumference drug,Hongzhongxiao tincture,on the treatment of skin abscess in rats under the guidance of defended field theory.Methods Forty SpragueDawley (SD) rats were randomly divided into normal control group,model group,western medicine control group,Hongzhongxiao pretreatment group and Hongzhongxiao treatment group according to the computer generated random number.Subcutaneous abscesses models were reproduced by subcutaneous injection of 1 mL (3-5 × 109 cfu/mL) of staphylococcus aureus,and the normal control group was subcutaneously injected with the same amount of normal saline.After molding,the rats in the model group were treated with normal saline for change of dressing;the western medicine control group was treated with mupirocin ointment for external use;Hongzhongxiao pretreatment group was treated with 2 mL of Hongzhongxiao for dressing 4 hours after the modeling;in Hongzhongxiao treatment group,2 mL dressing was given after the emergence of hard swelling,and the Hongzhongxiao drug range applied was 1 cm over the swelling range.Each group was dressed once daily.The changes of mental state,body weight,skin temperature,range and time of abscess formation,wound ulceration and healing time,transcutaneous oxygen pressure (TcPO2) at the periphery of abscess were observed in each group.Results ① After modeling,the activity of the rats was decreased,the appetite became worse,and the food intake less.Except the normal control group,the rats in other groups had different degrees of mental listlessness.② After 3 days of treatment,the body weight gain of the model group was significantly lower than that of the normal control group (g:8.75 ± 9.85 vs.31.67 ± 7.92,P ＜ 0.01),and continued to 18 days after treatment (g:27.13± 11.70 vs.98.00 ± 8.94);after treatment for 18 days,the body weight gain in the western medicine control group,Hongzhongxiao pretreatment group and Hongzhongxiao treatment group was significantlyhigher than those of model group (g:53.28 ± 19.69,49.12 ± 7.23,44.71 ± 12.42 vs.27.13 ± 11.70,all P ＜ 0.05).③ After 3 days of treatment,the body temperature of the model group was obviously lower than that of the normal control group (℃:33.75 ± 0.68 vs.35.03 ± 0.41,P ＜ 0.01) and continued to 10 days (℃:34.30 ± 0.35 vs.35.03 ± 0.41,P ＜ 0.01).The body temperature of the rats in Hongzhongxiao pretreatment group was significantly lower than that in the normal control group,the model group and the western medicine control group on the 14th day after treatment (℃:33.97 ± 0.83 vs.35.10 ± 0.57,35.01 ± 0.68,35.25 ± 0.23,all P ＜ 0.05).The body temperature of Hongzhongxiao treatment group was significantly higher than that of the Hongzhongxiao pretreatment group (℃:34.87 ± 0.94 vs.33.97 ± 0.83,P ＜0.05).④ After treatment for 13,15,17 days,the contraction rates of swollen area in western medicine control group,hongzhongxiao pretreatment group and hongzhongxiao treatment group were greater than the rate in model group,and the change was most significant on the 17th day after treatment [(96.37 ± 5.09)％,(92.76 ± 13.56)％,(98.41 ± 3.85)％ vs.(77.46 ± 19.07)％,all P ＜ 0.05].⑤ Compared with the model group,the times of abscess localization in the western medicine control group,the Hongzhongxiao pretreatment group and the Hongzhongxiao treatment group was shorter than that in the model group (days:9.28 ± 1.38,7.33 ± 1.97,7.67 ± 1.63 vs.12.63 ± 1.99,all P ＜ 0.05);the time of wound healing in Hongzhongxiao pretreatment group aud Hongzhongxiao treatment group were significantly shorter than those of the model group (days:17.67 ± 1.03,16.83 ± 1.51 vs.19.92 ± 2.33,P ＜ 0.05).⑥ TcPO2 in the Hongzhongxiao pretreatment group was significantly higher than that in the model group on the 7th day after treatment [mmHg (1 mmHg =0.133 kPa):63.33 ± 9.77 vs.39.51 ± 8.42],and the Hongzhongxiao treatment group (44.25 ± 6.41) was significantly lower than that of the Hongzhongxiao pretreatment group,and the Hongzhongxiao treatment group was significantly higher than that of the model group (59.50 ± 7.34 vs.49.52 ± 10.17) on the 14th day after drug application,and the western medicine control group was significantly lower than that of the model group (37.71 ± 5.63 vs.54.33 ± 7.74),the Hongzhongxiao pretreatment group and Hongzhongxiao treatment group were significantly higher than those of the western medicine control group,the difference being statistically significant (all P ＜ 0.05).Conclusions The Hongzhongxiao tincture applied on top and circumferentially around the periphery of the abscess can reduce the local inflammatory response,improve the body weight and mental state,increase the blood supply around wound periphery for anti-infection and anti-inflammation in order to promote the formation of defending field of nursing.Although the early application of Hongzhongxiao tincture cannot reduce the local infectious symptoms and kill the Staphylococcus aureus,it can make skin abscess localization as soon as possible and shorten the wound healing time.

Objective To investigate the clinical efficacy of our self-designed sight used to guide thoracic vertebroplasty.Methods A retrospective analysis was conducted of the 52 patients (70 thoracic vertebrae) who had undergone percutaneous vertebroplasty (PVP) (n =36)or percataneous kyphoplasty (PKP) (n =34) of T1-T4 vertebral bodies at Department of Orthopaedics,The First Affiliated Hospital to Zhengzhou University form August 2012 to October 2013.The operation time,intraoperative bleeding,intraoperative radiation by C-arm roentgenography,and visual analogue scale (VAS) were compared between the patients whose surgery had been guided by our self-designed sight and those whose surgery had been not.Results All the patients were followed up for 18 to 36 months (average,22.3 months).Compared with those who had not used the sight,the patients who had used the sight incurred significantly shorter operation time (16.5 ± 3.2 min versus 26.5 ± 3.7 min),significantly less intraoperative bleeding (2.8 ± 1.3 mL versus 6.3 ± 1.7 mL) and significantly less radiation by C-arm roentgenography (5.6 ± 3.3 times versus 9.4 ± 3.1 times) (P ＜0.05).The VAS scores at postoperative 3 days and final follow-up were significantly decreased than the preoperative values in all the patients (P ＜ 0.05).There were no significant differences between the patients who had used the sight and those who had not in the the VAS scores at postoperative 3 days or final follow-up (P ＞ 0.05).Conclusions The upper thoracic PVP or PKP guided by our self-designed sight has theadvantages of short operation time,less intraoperative bleeding and less frequency of C-arm radiation.The sight is suitable for various vertebroplasties.