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.

Objectives To explore the clinical significance of autoantibodiesin individuals who accept a routine physical examination.Methods From April to June 2012, the serum of 932 individualsincluding 649 males and 283 females, from department of routine physical examination center of Peking Union Medical College Hospitalwerecollected , it uesd IIF for ANA, line immunoassay ( LIA) for specific ANAs antibodies and ELISA for the other antibodies , includinganti-CCP antibodies , AMA-M2, ACL antibodies and anti-anti-β2GPⅠantibodies.Chi-square test was used for data measurement of positive rate of autoantibodies in men and women;Fisher′s exact test was used when the data not meet the conditions of Chi-square test.Individualswith high-risk of autoimmune disease according to the results ofautoantibodies ( Titersof autoantibodies≥2-fold cut-off and accompanied with other autoimmune diseases related laboratory abnormalities) were recalled to visit doctor.Results Of the 932 cases, the overall positive rate of ANA was 11.27%.The positive rate of ANA was19.79%inwomen, which was significantly higher than thatin men (7.09%)(χ2 =32.6, P<0.01); the overall positive rate of ANAswas 8.69%, and the positive rate of ANA was13.43%inwomen, whichwas significantly higher than that in men ( 6.63%) (χ2 =11.49, P <0.01);the overall positive rates of AMA-M2, anti-CCP antibodies , anti ACL antibodies and anti β2GPⅠantibodies were 3.22%, 0.54%, 2.90%and 0.21% respectively , which were 2.83%, 0.71%, 3.18%and 0.71 % in women , and 3.39%, 0.46%, 2.77% and 0.00% in men respectively , there was no statistically significant of positive rate between female and male 58 patients accounting for 6.22% in high-risk of autoimmune disease were recalled , of which 15 cases, accounting for 1.61% were diagnosed or highly suspected of autoimmune diseases (AID) of the 15 patients, 11 patients accounting for 1.18% were diagnosed AID, including 6 CTD, 3 pSS, 1 RA and 1 pSS/PBC;4 patients were highly suspected as AID , including 3 suspected CTD and 1 suspected pSS.The titers concentration of the positive antibodies in patients with confirmed or suspected AID ≥ 3 times cut-off.Conclusions The positive rate of autoantibodies in individuals of physical examination is high , but there is clinical significance when the titers concentration of positive autoantibodies ≥ 3 times of the cut-off.Positive-autoantibodies patients with high-risk of autoimmune disease need professional clinician to provide follow-up, consulting and health education for early discovery, timely diagnosis, and proper treatment of AID.

Objective To compare the clinical efficacy of the flaps of modified Limberg and direct incision followed by suture in the treatment of sacrococcygeal pilonidal sinus,aiming to provide the academic guide for its clinical surgery.Methods A total of 76 patients suffered from sacrococcygeal pilonidal sinus were treated with two surgical methods in Department of General Surgery and Department of Burn and Plastic Surgery in Sixth Medical Center of PLA General Hospital from 2013 to 2017.Among these patients,37 cases were resected and reconstructed using the flaps of modified Limberg,the other 39 cases were sutured after beingresected.These patients were followed up during the next 1 or 2 years.Five indexes including surgery period,length of hospital stay,postoperative infection,recurrence and complication,were investigated to statistically compare two different surgery methods' efficacy,using independent t test or chi-square test respectively.Results There were 76 patients in the Limberg flap group and the direct suture group,the operation time was about one and a half hours,and the hospitalization time was about 15 days (P＞0.05);there was no statistical difference between them.There were no complications in 35 of the 37 patients in the Limberg flap group,but skin cracking and infection in 2 patients.In the direct suture group,there were 39 patients,16without complications and 23 with complications,(x2=1133.2393,P=0.0000);there were statistical differences.Among the 37 patients in the Limberg flap group,only one recurred;among the 39patients in the direct suture group,9 recurred (x2=1192.9211,P=0.0000);there were statistical differences.Conclusions Limberg flap incision shows good healing and fewer complications.Within the group of Limberg flap incision healing,only few cases of recurrences occur after the surgery.It not only effectively reduce the complication rate in patients,but also release the pain from long-term,reduplicated medication.The author proposes that this operation should be promoted in the future clinical practice.

Objective This study aimed to assess the diagnostic value of anti-mutated citrullinated vimentin (MCV) antibodies in rheumatoid arthritis (RA) and its correlation with disease progression, extra-articular manifestations and overlap syndrome. Methods Retrospective Studies. Clinical data of 837 patients in PekingUnionMedicalCollegeHospitalfrom June to August 2017 were collected, including the result of anti-MCV, anti-cyclic citrullinated peptide (anti-CCP) antibodies, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR) and High-sensitivity-C-reactive protein (CRP). According to the 1987 American College of Rheumatology classification criteria for rheumatoid arthritis, there were 323 patients diagnosed with RA, including 59 males and 264 females with the average age of 51 years. According to whether the RA patients have overlap syndrome with other autoimmune disease (AID) or have extra-articular manifestations, 258 cases were categorized into RA group, including 47 males and 211 females with the average age of 50 years; 14 cases were categorized into the group of overlap syndrome, including 1 male and 13 females with the average age of 36 years;51 cases were categorized into the group of extra-articular manifestations, including 11 males and 40 females with the average age of 59 years.According to 2010 rheumatoid arthritis classification criteria for destruction in joints, the radiographic changes were divided into 4 stages. There were 203 casesenrolled in our study, 88 caseswere fitted into early stage group (stage I)including 21 males and 67 females with the average age of 48 years; 115 caseswere fitted into progressive stage group, which compromisedstageⅡ (interim stage), stage Ⅲ (severe stage) and stage Ⅳ(final stage) cases, including 19 males and 96 females with the average age of 53 years. Mann-Whitney U test, x2 test, Receiver operating characteristic (ROC) curves and Spearmancorrelation coefficientwere used in Statistical analysis. Results Ⅰ Amongdiagnosed RA patients, 199 (61.6％) cases were positive for anti-MCV, anti-CCP and RFsimultaneously, 42 (13％) cases were positive for anti-MCV, which was higher than anti-CCP positive (1 cases, 0.3％) or RF positive (7cases, 2.2％). The difference was statistically significant(P<0.001, P<0.001). ⅡROC was calculated and MCV=35.95 U/ml was used as best-fit cut-off value. The AUC for anti-MCV was 0.867, while the sensitivity was 80.5％and specificity was 80.9％.ⅢThe detection levels of anti-MCV (682.8 (106.4-1000.0)), anti-CCP (407 (4.0-1536.0)) and RF (82.8 (21.1-244.9)) in the group of progressive stage were higher than those in the group of early stage (114.5 (28.5-1000.0), 62.5 (5.0-1020.7), 50.1 (6.7-127.1)), which showed a significant difference(P<0.05, P<0.05, P<0.05). The anti-MCV, anti-CCP and RF were positively related to the degree of joint destruction (r=0.229, P<0.05;r=0.187, P<0.05;r=0.167, P<0.05);anti-MCV and anti-CCP were positively related to extra-articular manifestation (r=0.152, P<0.05;r=0.136, P<0.05). Conclusion Anti-MCV antibodies are more sensitive in patients with RA, and have complementary diagnostic value for anti-CCP and RF-negative patients; high levels of anti-MCV and anti-CCP in RA patients are associated with RA progression and extra-articular involvement.

Objective To investigate the efficacy of leflunomide combined with prednisone in the induction therapy of proliferative lupus nephritis (LN).Methods A prospective,multicenter,randomized controlled clinical trial was conducted in patients with biopsy-proved proliferative lupus nephritis recruited from 15 renal centers from 2013 to 2015.Patients were randomized to two groups.Oral leflunomide or intravenous cyclophosphamide was given to patients in each group.Both groups received a tapering course of oral prednisone therapy.All patients were followed up for 24 weeks.The blood biochemistry,urine index,clinical curative effect and adverse reaction were recorded and analyzed statistically.Results A total of 100 patients were enrolled in this clinical trial,including 48 patients in leflunomide group and 52 patients in cyclophosphamide group.After 24 weeks,the overall response rate was 79％ (95％ CI 67％-90％) in the leflunomide group and 69％ (95％ CI 56％-82％) in the cyclophosphamide group.23％ (95％CI 11％-35％) of patients in leflunomide group showed complete remission compared with 27％ (95％CI 24％-30％) in cyclophosphamide group (P=0.35).The levels of 24-hr urine protein excretion,SLEDAI and anti-dsDNA antibody titers were decreased in patients treated with leflunomide group after 24-weeks treatment.And the levels of serum albumin and complement 3 after treatment were significantly higher compared with these before treatment.There was also no significant difference in changes of 24-hr urine protein excretion,SLEDAI score,anti-dsDNA antibody titers,serum albumin and complement C3 levels after treatment between two groups.Incidence of adverse events did not differ between the leflunomide and cyclophosphamide group.Conclusions Leflunomide combined with prednisone showed same efficacy compared with cyclophosphamide as induction therapy for lupus nephritis.Leflunomide might be an useful medicine in the induction therapy of lupus nephritis.

To investigate the distribution and clinical significance of nuclear dense fine speckled (DFS) pattern in various diseases. A total of 95 289 patients who received DFS tests at Peking Union Medical College Hospital from January 2019 to December 2020 were included in this study. The results of indirect immunofluorescence assay (IIF) for detection of antinuclear antibody (ANA) were evaluated. The positive rates of ANA and DFS were 39.60% (37 733/95 289) and 1.19% (1 139/95 289) respectively. The positive rate of DFS in ANA-positive patients was 3.02% (1 139/37 733). DFS and ANA positivity were significantly different among different age groups rather than gender. The positivity rate of DFS reached the peak (55.57%, 633/1 139) in young patients between 21-40 years, while positive ANA with negative DFS was mainly observed in patients between 41-60 years (37.26%, 13 636/36 594). Additionally, single ANA-positivity were mainly detected in rheumatology department (59.23%, 18 402/31 066), whereas positive DFS was more common in obstetrics and gynecology department (3.08%, 49/1 593). There were 82.88% (944/1 139) patients with positive DFS diagnosed with non-autoimmune disease (non-AID), and 19.49%(222/1 139) with dermatosis. Positive DFS with higher titer (≥1∶320) was detected more frequently in autoimmune disease (AID) patients (5.13%, 10/195) than in non-AID patients (1.69%, 16/944) ( P<0.05). The DFS pattern is rare in ANA positive patients, which is mainly observed in women between 21-49 years. High titer of DFS is prevalent in AID patients, but positive DFS is detected more in non-AID patients, especially those with dermatosis.

The genetic background such as copy number, integration site and chromosome karyotype of exogenous genes of transgenic animals obtained by random integration is still unclear. There may be some problems such as silent integration, invalid integration, toxic integration and unpredictable expression level of exogenous genes. In this study, six primary (F0) and their corresponding offspring (F1) of human lactoferrin (hLF) transgenic goats were selected as the research objects, and blood samples were collected from jugular vein and DNA were extracted. The genetic background and expression level of exogenous genes were studied by chromosome karyotype analysis, real-time quantitative PCR (qPCR), ELISA and Western blotting. The chromosomes of six F0 transgenic goats had no obvious morphological variation, number change and other abnormalities. The relative copy number was different (2-16) and could be steadily inherited to the next generation. The copy number of F0 and F1 hLF gene was the same. The highest expression level of hLF was 1.12 g/L in F1 transgenic goats (L3-1, 8 copies). The results proved that the integrated exogenous genes could steadily inherit the next generation, and did not cause obstacles to the growth and development of transgenic goat individuals. Moreover, there was no obvious correlation between the number of copies and the expression level of hLF. This laid a foundation for the new varieties cultivation of transgenic goats and other transgenic animals, and analysis of genetic background.

China ; Female ; HIV Infections ; HIV-1 ; Humans ; Incidence ; Pregnancy ; Prevalence ; Sentinel Surveillance

OBJECTIVETo investigate the distribution of hepatitis C virus (HCV) subtypes among intravenous drug users (IDUs) in Kunming, Yunnan province, in 2014.

METHODSA total of 276 plasma samples were collected from IDUs in Kunming during April 2014 -July 2014, in which 199 plasma samples were tested to be HCV antibody positive. For the HCV antibody positive samples, HCV E1E2 genes and NS5B genes were amplified by using nested-polymerase chain reaction (PCR). After sequencing, the HCV subtypes were determined through phylogenetic analysis.

RESULTSBased on the phylogenetic trees of E1E2 gene and NS5B gene fragments, a total of 125 samples were genotyped. 3b was the predominant subtype (48.8%,61/125), followed by 3a (30.4%, 38/125), 6n (14.4%, 18/125), 6a (3.2%, 4/125) and 1b (3.2%, 4/125). The distributions of HCV subtypes by sex, marital status, ethnic group and HIV-1 infection status showed no statistical differences. However, the distribution of HCV subtypes by age showed statistical difference, the diversity of HCV subtypes was found in age group <45 years. The genetic distances of 3a, 3b and 6a on E1E2 and NS5B were larger than those of 1b and 6n. For genotype 3, the genetic distances of 3b on E1E2 and NS5B were larger than those of 3a.

CONCLUSIONFive HCV subtypes were found among IDUs in Kunming, HCV subtypes 3b and 3a were predominant, which have circulated in this population for long time.