A proteoglycan-induced spondyloarthritis mouse model and the therapeutic effect of CCN1 monoclonal antibody / 中华风湿病学杂志

Objective To set up a mouse model of spondyloarthritis,analyzethe clinical phenotype,radiographic and pathological features,and investigate the therapeutic effect of cysteine-rich 61 (CCN1) monoclonal antibody in spondyloarthritis mouse model.Methods Proteoglycan from bovine nasal septum was used for immunization of 14-16 week old female BALB/c mice.CCN1 monoclonal antibody 093G9 or control immunoglobulin (Ig)G were injected to the spondyloarthritis mice.The arthritis scores were analyzed by t test.Peripheral and axial joints disease development was assessed by Micro-CT and histology.Results Proteoglycan immunized mice began to develop peripheral arthritis in the 8th week.The peripheral arthritis score reached the peak (10.5±1.5) in the 11th week,with the inflammation and spur formation of the ankle and knee joint.We found infiltration of inflammation cells in intervertebral discs of the lumbar vertebrae and the caudal vertebrae.Chondrocyte proliferation couldbe seen in the meniscus of knee and lumbar intervertebral discs.In the 18th week,the intervertebral discsof thoracic vertebrae and the cervical vertebrae were also damaged.Abundant chondrocytesgathered in the intervertebra] discs.The inflammation and new bone for-marion of peripheral and axial joints were more severe in control IgG group than 093G9 group.The peripheral arthritis score in the 093G9 group decreased significantly after 2 treatments,[(2.8±1.3) vs (4.2±2.1),t=2.516,P＜0.05].The difference in arthritis scores between the two groups was the most significant after 8.treatments,[(2.0±2.0)vs (5.3±2.0),t=4.082,P＜0.01].Conclusion The mouse model of spondyloarthritissimulates human spondyloarthritis,including inflammation and new bone formation in p()gheral and axial joints.CCN1 monoclonal antibody can improve the inflammation and new bone formation inspondyloarthritis mouse model.

The experimental study on the effect of rhTNFR:Fc and methtotrexate-rhTNFR:Fc on joint destruction of collagen-induced arthritis rat / 中华风湿病学杂志

Objective This study is aimed to explore the effect of rhTNFR:Fc and methotrexate (MTX)-rhTN FR:Fc on joint destruction of collagen-induced arthritis ( CIA ) rat by establishing CIA rat model which imitates pathogenic factors of rheumatoid arthritis (RA).Methods CIA rat model were developed by subcutaneous injection of bovine type Ⅱ collagen.The rats with inflammation scores of two or above were randomly divided into four groups:the sterilized water treatment group (0.4 ml/w,intra-peritoneal injection),the MTX treatment group (1 mg/w,intra-peritoneal injection),the rhTNFR:Fc treatment group(0.8 mg Biw,intra-peritoneal injection),the MTX + rhTNFR:Fc treatment group (MTX 1 mg/w and rhTNFR:Fc 0.8 mg Biw,intraperitoneal injection).After treatment for 8 weeks,the rats were sacrificed and took the ankle radiography.Micro-CT scan of proximal tibia was performed and hard-tissue slices were made,and then the ankle's bone damage of each group was observed in order to evaluate trabecular variation and bone quantity changes of proximal tibia.Statisstical analysis was conducted with ANK-q test.Results After treatment for 8 weeks,the percentage of trabecular area and the trabecular number of the rhTNFR:Fc treatment group and the MTX-rhTNFR:Fc treatment were [(29.1±0.3)％,(26.7±0.6)％,(4.4±0.5)/mm,(4.0±0.6)/mm] (P＜0.01),which were evidently higher than the sterilized water treatment group and MTX treatment group (P＜0.01).The trabecular separation of Etanercept treatment group and MTX-rhTNFR:Fc group was obviously less than the sterilized water treatment group and MTX treatment group [(12.9±0.5)％,(13.2±0.4)％ vs (2.0±0.3)/mm,(2.2t0.2)/mm] (P＜0.01).Conclusion rhTNFR:Fc and MTX-rhTNFR:Fc can remarkably inhibit joint destruction of CIA rat.And their effect on inhibiting of inflammation and increasing peri-articular bone quantity.In addition,they are effective on inhibiting the reduction of local trabecular structure and increase of trabecular separation.

Cyr61 promotes proliferation of fibroblast-like synoviocytes in rheumatoid arthritis and its regulation by inflammatory factor / 中国免疫学杂志

Objective:To investigate the effect of Cyr61 on the proliferation of Fibroblast-like Synoviocytes (FLS) in rheumatoid arthritis (RA).Methods:Cyr61 expression in synovial tissues (ST) and FLS was examined using Real-time PCR,Western blot and immunohistochemistry simultaneously.FLS were isolated from synovial tissue of RA patients and cultured in vitro.The proliferation of FLS stimulated with synovial fluid (SF) was determined by 3 H-TdR incorporation.Cyr61 protein level in RA SF was detected by ELISA.Results:Cyr61 was over expressed in ST,FLS of RA patients while hardly examined in normal individuals and osterarthritis (OA) patients.Meanwhile,Cyr61 protein level was elevated in SF,which promoted the proliferation of RA FLS.This proliferation was abrogated by knockdown the Cyr61 gene of FLS or neutralizing monoclonal antibody against human Cyr61.Moreover,inflammatory factor IFN-γ and TNF-α up-regulated the expression of Cyr61.Conclusion:These results indicate that the elevated level of IFN-γ and TNF-α in RA SF can promote the proliferation of the FLS derived from RA patients by increasing expression of Cyr61,suggesting that Cyr61 may play an important role in the development of RA.

Mechanism of Treg down-regulation by immunization with attenuated activated autologous T cells / 上海交通大学学报（医学版）

Objective To explore the mechanism of down-regulation of regulatory T cells (Treg) by immunization with attenuated activated autologous T cells. Methods Aulologous T cells were activated with ConA in vitro. Mice were immunized subcutaneously and inlraperitoneally every 5 days for 3 times (5 ×10~6 per time for each mouse), and the number and function of Treg were examined. PBS was subcutaneously injected for control group. Serum level of anti-mouse CD25 antibody was measured by ELISA. The number and function of Treg was detected by serum adoptive transfer and proliferation and inhibition assays. Results Compared with control group, there were less CD4~+ CD25~+ Foxp3~+ Treg in the mice after immunization (P < 0. 01), the immunosuppression ability decreased (P<0. 01), and the level of anti-CD25 antibody increased (P <0.01). Adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice (P<0. 01). Conclusion Immunization with attenuated activated autologous T cells induces more anti-CD25 antibody, which may further down-regulate CD4~+CD25~+Foxp3~+ Treg expansion and function in vivo.

Mouse bone marrow derived mesenchymal stem cells suppress lymphocyte proliferation through co-Inhibitor B7-H1 upregulated by IFN-γ / 中华微生物学和免疫学杂志

Objective To study the mechanism of mesenchymal stem cells immunosupression lym- phocyte proliferation via B7-H1/PD-1 pathway upregnlated by IFN-γ. Methods Bone marrow derived mes-enchymal stem cells (MSC) were isolated and purified by repeat adherent passage and detected them multi-potential differentiation in conditioned culture medium. Then MSC were cocuhured with lymphocyte prolifera-tion and assayed the level using γ H-thymidine incorporation. Meanwhile, ELISA measured IFN-γ, TGF-β, TNF-α and IL-10 in the cocuhured supernaatant and analyzed variation of B7-H1 molecular profile in MSC by flow cytometry. At last siRNA technology was deploied to interfere B7-H1 expression and analyzed MSC im-munosuppression on lymphocyte proliferation. Results In vitro the isolated MSC become homogeneous spi-die-shaped adherent cells after five passages, and in conditioned culture medium they could differentiate into adipocytes, osteocytes and chondrocytes. In the eocuhure of MSC with mixed lymphocyte, lymphocyte prolif- eration stimulated by Con A or by anti-CD3/CD28 antibody. The cpm value of the proliferation detected by 3H thymidine incorporation showed MSC suppressed the proliferation significantly (P = 0. 0167, 0. 0081,<0.0001 ) and the suppressive potential in a dose-dependent fashion. In the coeuhured supernatant cyto-kine IFN-T and TNF-α were detected in high concentration, but TGF-β, IL-10 were undetected. Simultane- ously MSC in the coeuhure upregulated B7-H1 expression from basic expression 7% to higher than 70% ( P < 0.05 ). After interfere B7-H1 expression in MSC by specific siRNA, we detected lymphocyte proliferation and got higher cpm by 3H thymidine incorporation ( P < 0. 05 ). Conclusion MSC upregulated B7-H1 mo-lecular expression upon the stimuli of IFN-γ, and through the B7-H1/PD-1 pathway mediated immunosu-pression on lymphocyte proliferation.

Comparison of transperitoneal and extraperitoneal laparoscopic radical prostatectomy / 中华泌尿外科杂志

Objective To compare the clinical features and results between transperitoneal laparoscopic radical prostatectomy and extraperitoneal laparoscopic radical prostatectomy.Methods Thirty-three prostate cancer patients treated with laparoscopic radical prostatectomy. Among them,21 cases had been done transperitoneally and 12 cases had been done extroperitoneally. The two different approaches were evaluated and compared in respects of operating time, estimated blood loss, complications during surgery, postoperative complications, intestinal function recovery time, catheterization time and length of hospital stay.Results All the surgeries had completed successfully without conversion to open surgery. For transperitoneal approach and extraperitoneal approach, the operating time was (299±46)min and (309±64)min, blood loss was (618±448)ml and (677±469)ml. There were 3 cases with severe blood loss, 2 cases with bladder injuries and 1 case with ureteral injury in transperitoneal approach group. There were 1 case with severe blood loss, 1 case with obturator never injury, 1 case with cysto-ureteral injury and 1 case with peritoneum injury in extraperitoneal approach group. For transperitoneal approach and extraperitoneal approach,the catheterization time was(14.6±3.8)d and (12.3±2.9)d, intestinal function recovery time was (2.7±0.7)d and (2.1±0.5)d, length of hospital stay was (17.0±3.6)d and (11.2±3.5)d, respectively.Conclusions Laparoscopic radical prostatectomy is feasible and safe in clinical practice. Extraperitoneal approach has better vision, less impact on abdominal organs, faster recovery and shorter hospital stay comparing to transperitoneal approach.

Extra-adrenal pheochromocytoma: report of 34 cases / 中华泌尿外科杂志

Objective To review the diagnosis and treatment of extra-adrenal pheochromocytoma with a 34cases report. Methods Thirty-four cases of extra-adrenal pheochromocytoma were retrospectively analyzed. Hypertension was observed in 27 cases. Abdominal pain was seen in 10 patients and intermittent hematuria in 2 patients. Serum and urinary catecholamine and urinary VMA were measured in 34 cases. The level of serum or urinary catecholamine elevated in 20 cases and urine VMA elevated in 24 cases. Thirty-four cases had ultrasound examination,25 cases underwent CT scan and 6 cases underwent MER scan.Results Pheochromocytomas of 12 cases were located in the renal hilum, 2 in the lower pole of the left kidney, 1 in the posterior aspect of the inferior vena cava, 3 in the interaortocaval region, 2 in the anterior aspect of the abdominal aorta, 1 in the anterior of the right common iliac artery, 1 in the hilum of the liver, 1 in the posterior o{ the pancreas, 2 in the bladder wall, 1 in the posterior of the descending colon, and 8 cases of multifoci. Twenty-two cases of extraadrenal pheochromocytoma were benign and 12 cases were malignant. Thirty cases were followed up from 6 months to 13 years. Among 27 cases with hypertension, the blood pressure of 22 patients returned to normal and 5 cases were still hypertensive. Nine cases of malignant pheochromocytoma all had tumor recurrence or metastases at one year postoperatively. Six patients died during followed-up from 6 months to 3 years, including 3 cases died of cerebral hemorrhage and 3 cases of tumor metastases. Three cases got stable with 131Ⅰ-MIBG radiotheraphy.Conclusions The accurate detecting extra-adrenal pheochromocytoma is difficult. CT scan could be reliable in localizing the lesions. Surgical resection of the tumor could be the best therapy. Patients of malignant extra-adrenal pheochromocytoma may be treated with 131Ⅰ-MIBG after surgical therapy.

Clinical analysis of 15 cases of Paget's disease of the scrotum / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features and treatment of Paget's disease of the scrotum.</p><p><b>METHODS</b>Fifteen cases of Paget's disease of the scrotum, pathologically diagnosed and successfully treated by surgery from 1987 to 2003, were studied retrospectively.</p><p><b>RESULTS</b>Typical pathological features were observed in all the patients. Thirteen cases were followed up for 0. 5 approximately 13 years, of whom 1 case relapsed within 3 years, 2 died of metastasis after 2 years, and 3 died of cardiovascular diseases, with no recurrence in the others.</p><p><b>CONCLUSION</b>Early biopsy and extensive excision of the scrotal lesion is the first choice of treatment for Paget's disease, with good operative effect and satisfactory prognosis. A 2nd operation could be considered for patients of local relapse.</p>

Effect and mechanism of alpha1-adrenoceptor blocker combined with antibiotics for chronic prostatitis / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism of alpha1-adrenoceptor blocker combined with antibiotics in the treatment of chronic prostatitis.</p><p><b>METHODS</b>Eighty patients with chronic prostatitis were divided into two groups, one treated with alpha1-adrenoceptor blocker (Terazosin 2 mg qn) and Levo-ofloxacin (0.2 bid), and the other given Levo-ofloxacin (0.2 bid) alone for 6 weeks. Chronic prostatitis symptom index (CPSI), urodynamic data and prostatic secretion examination were compared before and after treatment.</p><p><b>RESULTS</b>The CPSI score of the treated group decreased from 31.8 +/- 7.4 to 15.5 +/-6.6, while that of the control group decreased from 30.9 +/- 7.1 to 21.4 +/- 6.2. There was significant difference between the two groups (P < 0.05). The maximum flow rates before and after the combined treatment were 16.5 +/- 6.3 ml/s and 20.4 +/- 4.6 ml/s, while those before and after Levo-ofloxacin administration were 16.1 +/-5.8 ml/s and 17.3 +/- 6.8 ml/s. The difference was significant (P < 0.05). The maximum urethral pressure of the combined treatment group decreased from 92.5 +/- 15.3 cm H2O to 72.5 +/- 13.4 cm H2O, while that of the control group decreased from 93.2 +/- 14.8 cm H2O to 91.7 +/- 13.6 cm H2O.</p><p><b>CONCLUSION</b>Alpha1-adrenoceptor blocker can lower the intraurethral pressure, which prevents urine from refluxing to the prostate. Alpha1-adrenoceptor blocker combined with antibiotics is effective for chronic prostatitis.</p>

Study on a type Ⅱ collagen-specific T cell line and the pathogenesis of arthritis / 中华风湿病学杂志

Objective To establish the type Ⅱ collagen specific T cell line of Wistar rat and observe its effect on transferring arthritis.Methods The Wistar rats were immunized with emulsified chicken type Ⅱ collagen (CCⅡ) in complete Freund′s adjuvant by intradermal injection to induce the rat model of collagen induced arthritis (CIA).The lymphocytes were obtained from mesenteric lymph nodes of CIA rats,and the type Ⅱ collagen reactive T cell line was selected and propagated by CCⅡ stimulating in vitro .The proliferation response and phenotype were analyzed by 3 H TdR incorporation and fluorescence activated cell sorter (FACS).The onset of arthritis and pathological characteristic in ankle joints of recipient rats were observed with naked eye and histochemical examination.Anti CCⅡ antibody in serum was assayed by enzyme linked immunosorbent assay (ELISA).Results A T cell line was successfully established.The results of FACS labeled with fluorescent antibodies showed that 98 2% of the line cells were T cells,of which 89 7% were CD4 + T cells.The results of adoptive transfer showed that the incidence of arthritis was 50% when the injected cell number was 5?10 7,meanwhile the level of anti CCⅡ antibody in serum was elevated more than that of the control.Conclusion A cell line has been successfully established.The result of arthritis transferring by T cell line shows that the T cell plays a great role in the pathogenesis of CIA and provides a research datum for rheumatoid arthritis therapy with T cell vaccine.

Activation and clonal expansion of T cells in the synovial fluid and peripheral circulation in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the difference between T cells in the synovial fluid and peripheral blood in patients with rheumatoid arthritis(RA). Method Samples from 22 patients were studied. The differentiation and activation markers expressed on T cell surface were detected by immunofluorscence using flow cytometer. The specific proliferation of collagen Ⅱ and heat shock protein 70 was analyzed using standard 3H-TdR incorporation method. Restricted V beta usage of these T cell was analyzed by semi-quantitied RT-PCR. Results The majority of the T cell subsets in the synovial fluid were demonstrated to be CD4 and CD8 positive cells in which (40?10)% were CD4 positive and (36?16)% were CD8 T cells respectively. The ratio between CD4 and CD8 was much lower than that found in the PBL of RA patients. The percentage of CD3+/CD25+ T cells was (16?6)%. The specific proliferation of collagen Ⅱ and HSP70 to CD3+/CD25+ T cell was higher than that of CD3+/CD25+ negative T cells. The T cell receptor expressed on the T cells from both peripheral blood and synovial fluid were tested for ?? TCR (70?26)%. However, the T cells in the synovial fluid showed V?14,16 and 17 restriction. Conclusion The data here reported indicates that T cell subsets in the synovial fluid and peripheral blood circulation in patients with rheumatoid arthritis are different. The T cells in the synovial fluid demonstrates more activation and higher reactivation to collagen Ⅱ and HSP70. The TCR of T cells showes V?14,16 and 17 restriction.

Immune suppression induced by auto-keratinoc-ytes in intermingled skin grafting / 细胞与分子免疫学杂志

Aim To explore the mechanism of the immune tolerance induced by the auto-islets in the intermingled skin grafting. Methods The mixed epidermal cell and lymphocyte culture (MELC) was established, in which auto-keratinocytes were added to mimic the effect exerted by the auto-islets in vivo. Results Auto-keratinocytes could inhibit the lympho-proliferation induced by alloantigen. Conclusion The immune tolerance induced by auto-islets may be related to the function of auto-keratinocytes as antigen presenting cells.

Treatment of urinary incontinence with artificial urinary sphincter (report of 4 cases) / 中华泌尿外科杂志

Objective To explore the efficacy and safety of artificial urinary sphincter in the treatment of real urinary incontinence. Methods The implantation of AMS 800 sphincter prostheses was conducted on 4 patients with urinary incontinence.The voiding diaries were recorded and urodynamic examinations were done before and after the implantation.The adverse reactions were recorded after the procedure. Results In the 4 cases,the sphincter pump was activated 4 to 6 weeks after the implantation.The urinary incontinence was well controlled and automatic urination was recovered.No infection-related complication leading to failure of the artificial urinary sphincter occurred.Follow-up for 17 to 46 months showed no leakage of urine at daytime or at night. The mean score of quality of life was 1.Urodynamic examination showed satisfactory compliance and stability of the bladder.The urethral pressure profiles (UPPs) being 8.2 to 9.4 kPa (1 kPa=10.20 cmH 2O);and the mean was 8.7 kPa.Three cases had no residual urine,while only 1 case had 60 ml of residual urine.Only 1 patient had to change the control pump due to mechanical failure 27 months after the operation. Conclusions Artificial urinary sphincter implantation is an effective and safe therapy for real urinary incontinence.The proper use of AM 800 and improved technique result in good long-term outcomes.

Inverted urothelial papilloma (report of 113 cases) / 中华泌尿外科杂志

Objective To evaluate the biological behavior,clinical features,and diagnosis and treatment of inverted urothelial papilloma. Methods A total of 113 cases of urothelial inverted papilloma were included in this retrospective analysis.Of them 102 were male and 11 were female,with a mean age of 55 years.Among the 113 cases of urothelial inverted papilloma 108 had the papilloma at low urinary tract.Of these cases,88 were treated by TURBT,6 by TURBT and TURP,10 by partial cystectomy,4 by partial cystectomy and prostatectomy,1 by total cystectomy.Five cases who had the papilloma at upper urinary tract underwent nephroureterectomy. Results 86 cases were followed up 0.5 to 10 years.Intravesical recurrentce occurred in 5 cases.Of them 2 cases developed malignance and 1 case underwent total cystectomy. Conclusions Inverted urothelial papilloma is a benign tumor,which appears male predominant.Most of the lesions localize at the bladder.The diagnosis can be established mainly by cystoscopy.Most of the inverted urothelial papilloma can be cured by TURBT with a satisfactory prognosis.

Autologous buccal mucosal patch grafting in the surgical treatment of Peyronie's disease / 中华泌尿外科杂志

Objective To evaluate autologous buccal mucosal patch grafting for correcting penile deformity in the surgical treatment of Peyronie's disease. Methods 14 patients with symptoms of Peyronie's disease for more than three months suffered from persistent pain ,severe curvature of the penile shaft and resistant to conservative treatment underwent plaque incision and autologous buccal mucosa patch grafting. Results Postoperative follow up has been 0.5 to 2years.The penile shaft became straight with no narrowing or indentation in 12 patients (86%) . In 14% of cases (2/14) some curvature of the penile shaft persisted and there was pain on erection.In all the patients there was no change in penile length. 86% of patients had overall satisfaction after surgery with improvement in psychological and social well-being. Conclusions The autologous buccal mucosal patch graft provides an anatomical and functional tunied substitute resulting in correction of the penile deformity in Peyronie's disease.

Molecular cloning and characterization of new SLA DQA alleles and their significance / 中国免疫学杂志

Objective:To lay background for studying rejection mechanisms in xenotransplantation, class II DQA genes of swine leukocyte antigens (SLA) of three Chinese pig strains GXP, GZP and YNP were cloned and sequenced.Methods:RT-PCR was performed to proliferate SLA-DQA genes (GXPDQA, GZPDQA and YNPDQA),which were cloned into pGEM-T Easy vector for sequencing and analysis.Results:All three allelic sequences examined, are not identical to those reported, which allows the sequences receiving their accession numbers from GenBank as AY102473, AY102474 and AY102475, which encompass an open reading frame of 765, 768 and 768 nucleotides respectively.Conclusion:Three new alleles of SLA-DQA genes were obtained. Analysis of the amino acid sequences of swine, human and mouse DQA (or equivalent) also indicated that GXPDQA, GZPDQA and YNPDQA were more similar to human DQA than mouse H-2-A?, which might provide an understanding of how the immune system evolved.