Andrology in China： current status and 10 years＇ progress

Andrology has a long history in traditional Chinese medicine. There are records of male sexual health, male sexual dysfunction and male infertility from over thousands of years ago. Modern andrology in China had a late start, with the Chinese Androlo~ Association founded in 1995. Within last decade, andrology in China has grown rapidly. In this review article, we summarized the progress of androlo~ in last 10 years and outlined the current status of Chinese andrology with a special focus on progress in male erectile dysfunction, prostate diseases, male infertility and male hormonal contraception.

Advanced primary peritoneal carcinoma:clinicopathological and prognostic factor analyses

Objective: To investigate the factors favoring a positive prognosis for advanced primary peritoneal carcinoma (PPC). Methods: Twenty-four cases meeting the criteria for PPC were analyzed retrospectively for the clinicopathologic profiles. Im- munohistochemistry was used to determine the expressions of p53, Top2α, Ki-67 and Her-2/neu. Then all these clinicopa- thological factors and molecular markers were correlated with the prognosis. Results: There were 15 cases of primary peritoneal serous papillary carcinoma (PPSPC), 6 cases of mixed epithelial carcinoma (MEC) and 3 cases of malignant mixed Mullerian tumor (MMMT). All patients underwent cytoreductive surgery with optimal debulking achieved in 3 cases. Among those re- ceiving first-line chemotherapy, 13 patients received the TP regimen (paclitaxel-cisplatin or carboplatin) and 7 patients received the PAC regimen (cisplatin-doxorubicin-cyclophosphamide). The median overall survival of all patients was 42 months, while the breakdown for survival time for patients with PPSPC, MMT and MEC was 44, 13 and 19 months, respectively. The expressions of p53, Top2α and Ki-67 were all demonstrated in 11 cases respectively. None showed the expression of Her-2/neu. There were significant differences in the median survival between patients with PPSPC and those with MMMT (44 months vs 13 months, P<0.05), also between patients receiving TP combination and those receiving the PAC regimen (75 months vs 28 months, P<0.05). Another significant difference in the median progression-free survival (PFS) was identified between patients with positive p53 immunostaining and those with negative p53 immunostaining (15 months vs 47 months, P<0.05), whereas age, menopausal status, residual tumor size and the other molecular factors did not significantly impact survival. Conclusion: Patients with PPC should be treated with a comprehensive management plan including appropriate cytoreductive surgery and responsive chemotherapy. Overestimating an optimal debulking surgery may not benefit survival. The pathologic subtype, chemotherapy regimen and p53 overexpression were significant prognostic factors.

Progress in gene transfer by germ cells in mammals

Use of germ cells as vectors for transgenesis in mammals has been well developed and offers exciting prospects for experimental and applied biology, agricultural and medical sciences. Such approach is referred to as either male germ cell mediated gene transfer （MGCMGT） or female germ cell mediated gene transfer （FGCMGT） technique. Sperm-mediated gene transfer （SMGT）, including its alternative method, testis-mediated gene transfer （TMGT）, becomes an established and reliable method for transgenesis. They have been extensively used for producing transgenic animals. The newly developed approach of FGCMGT, ovary-mediated gene transfer （OMGT） is also a novel and useful tool for efficient transgenesis. This review highlights an overview of the recent progress in germ cell mediated gene transfer techniques, methods developed and mechanisms of nucleic acid uptake by germ cells.

Magnetic Resonance Urography and X-ray Urography Findings of Congenital Megaureter

Objective To observe the imaging findings of congenital megaureter in order to enhance the understanding of this disease. Methods Image data of 5 patients with congenital megaureter and 2 misdiagnosed patients were analyzed, and image findings of congenital megaureter were summarized. Elscint Prestig 2.0T superconductive magnetic resonance urography (MRU) with conventional sequence (spin-echo, T1WI 560 ms/16 ms; fast spin-echo, T2WI 9600 ms/96 ms) was performed. Raw data were acquired with fast spin-echo sequence from heavy T2-weighted image (9600 ms/120 ms). Post-processing method of MRU was the maximum intensity projection with three-dimensional reconstruction in the workstation. Intravenous pyelography (IVP) was conducted, in which X-ray films were taken 7 minutes, 15 minutes, and 30 minutes after injecting contrast agent, except that in 2 patients the films were taken delayed at 60 and 90 minutes. X-ray retrograde pyelography was performed on 2 patients, successful in one but failed in the other. Results The dilated ureter showed hypointensity on T1-weighted images and hyperinten-sity on T2-weighted images in conventional MRI. The mass wall was intact, uniform in thickness, and showing hypointensity on T1-weighted and T2-weighted images. The MRU images showed a retroperitoneal mass appearing as an elongated tubular cystic structure spreading from kidney to bladder. MRU also revealed dilated calices and renal pelvis, pelviureteric obstruction, and renal duplication. The main signs of congenital megaureter in X-urography was significant dilatation of ureter, or normal renal pelvis with ureter dilatation, hydronephrosis, deformity, and displacement. Conclusions MRU with X-urography could visualize the characteristics of congenital megaureter, including the dilation of renal pelvis and ureter, calculi, urinary tract duplication, and stenosis location. The two techniques can complement each other in disease diagnosis and pro-vide more detailed information for preoperative treatment.

Early and delayed castrations confer a similar survival advantage in TRAMP mice

The most appropriate time to introduce androgen deprivation therapy for prostate cancer remains controversial. Our aim was to evaluate the effects of early versus delayed surgical castration on prostate cancer progression and survival in the transgenic adenocarcinoma of the mouse prostate （TRAMP） model. TRAMP mice were randomly divided into three groups： the early castration group （on which castration was performed at the age of 4 weeks）, the delayed castration group （on which castration was performed when abdominal tumours could be palpated）, and the sham-castrated group. Mice were monitored daily throughout their lives until cancer-related death or the develop- ment of an obviously moribund appearance, at which time the individual mouse was killed. Androgen receptor expression in prostate tumours was also evaluated. The results shows that the average lifespan in early castration, delayed castration and sham-castrated groups were 54.1 weeks, 59.9 weeks and 39.1 weeks, respectively. Both early castration and delayed castration conferred a statistically significant survival advantage when compared with the sham-castrated group （P 〈 0.001）. However, the difference in lifespan between the early castration group and the delayed castration group was not statistically significant （P = 0.85）. The increase in lifespan in the TRAMP mice that received either early or delayed castration correlated with lower G/B value （genitourinary tract weight/body weight） at death than the sham-castrated mice. In conclusion, early and delayed castrations in TRAMP mice pro- longed survival to a similar extent. This finding may provide a guide for clinical practice in prostate cancer therapy.

来源于人膀胱癌细胞株EJ的HSP70负载的DC所激发的特异性CTL细胞的体外应答效应(英文)

Objective: To investigate whether human dendritic cells (DC) derived from peripheral blood mononuclear cells (PBMC), which were pulsed by heat shock protein 70 (HSP70) isolated from human bladder tumor cell lines of EJ, were able to induce peptide specific cytotoxic T-lymphocytes (CTL) response in vitro and give the experimental foundation for the future clinical trials of immunotherapy in bladder tumor. Methods: The EJ-derived HSP70 co-cultured with DC from the healthy volunteers' PBMC, along with the crude lysate (the supernatant before HSP70 purification) from EJ cells were used as the experimental groups and DC not pulsed by any tumor cells antigen were the blank control. The autologous T-lymphocytes were added into the above various DC groups, and after incubation, the stimulation indexes (SI) and interferon-γ (IFN-γ) were detected to evaluate the immune activities of various DC groups. The killing effects of CTL to target cells, EJ and Hela cells, were determined with 51Cr releasing test. Results: Both DC/HSP70 and DC/the crude lysate could effectively activate CTL in vitro and kill target cells EJ. The killing effect of DC/HSP70 to EJ was much stronger than DC/the crude lysate (the supernatant before HSP70 purification) (P < 0.05). DC without any tumor cell antigens had a lower killing power to EJ. Meanwhile, DC/ HSP70 had little killing power to Hela non-relevant to bladder tumor histopathologically as compared with EJ cells (P < 0.05). Conclusion: The DC pulsed by HSP70 derived from the autologous tumor cells could induce a peptide complexes specific CTL response to tumor cells, and the CTL response induced by the DC/HSP70 was stronger, which display the basis of the possible clinical application of DC/HSP70 for bladder tumor.

Paraneoplastic Syndromes of Hypercalcemia and Leukocytosis Associated with Colonic Metastases from Squamous Cell Carcinoma of the Lung: a Case Report

Lung cancer is the most common cancer-related death in both men and women in the world. Approximately 25% of all cancer deaths are attributable to lung carcinoma. Moreover, about one-half of patients with lung cancer have metastases at the time of initial diagnosis, most frequently of lymph nodes, adrenals, liver, bone and brain.

Hemorrhagic cystitis following hematopoietic stem cell transplantation:incidence,risk factors and association with CMV reactivation and graft-versus-host disease

Background The definite pathogenesis of hemorrhagic cystitis （HC） after allogenic hematopoietic stem cell transplantation （allo-HSCT） has not been well elucidated. The role of cytomegalovirus （CMV） reactivation and graft-versus-host disease （GVHD） in the development of HC remains obscure. This study determined the incidence and risk factors for HC after alIo-HSCT and analyzed its association with CMV reactivation and GVHD. Methods We retrospectively studied 250 patients at high risk for CMV disease who underwent alIo-HSCT all based on busulfan/cyclophosphamide （BU/CY） myloablative regimens. The incidence, etiology, risk factors and clinical management of HC were investigated. Results HC developed within 180 days of transplant in 72 patients, with an overall incidence of 28.8% and an incidence of 12.6% in patients with HLA-matched related donors （MRD）, 34.38% in those with HLA-matched unrelated donors （MUD）, 49.45% in those with mismatched related donors （MMRD）. CMV-viremia significantly increased the incidence of later onset HC （LOHC）; however, only 9 out of 15 patients with CMV viruria actually developed LOHC. Multiple regression analysis identified grade II-IV acute GVHD （RR=2.75; 95% CI 1.63-4.66; P〈0.01） and grafts from MUD or MMRD （RR=2.60; 95% CI 1.52-5.20; P〈0.01） as independent risk factors for HC. Event sequence analysis indicated a majority of HC episodes began around GVHD initiation. Conclusions CMV-viremia is a high risk factor for LOHC. Our data also showed a correlation between acute GVHD and HC, which suggested that alloimmunity may be involved in the pathogenesis of HC.

High-normal serum uric acid is associated with albuminuria and impaired glomerular filtration rate in Chinese type 2 diabetic patients

Background Recently, some studies had shown that elevated serum uric acid （SUA） itself may increase the risk for development of renal disease in patients with diabetes. This study aimed to explore whether SUA was a predictor of microalbuminuria and impaired renal function in type 2 diabetes in Chinese patients. Methods This cross-sectional study included 2108 type 2 diabetic patients. Kidney function was estimated using the simplified modification of diet in renal disease （MDRD） equation to obtain estimated glomerular filtration rate. The urine samples were obtained for measuring the albumin-to-creatinine ratio （ACR）. Results According to the ACR level, these patients were divided into two groups, normal ACR （NA） and non-normal ACR （non-NA）. Both SUA and creatinine were significantly higher in the non-NA group than those in the NA group （（318.89±107.52） vs. （283.44±88.64） pmol/L, and （95.08±53.24） vs. （79.63±18.20） μmol/L, respectively）. Logistic regression analysis showed that diabetic duration, systolic blood pressure, creatinine and SUA were the independent predictors of albuminuria. Furthermore, to identify the factors associated with renal function, these patients were divided into two groups according to the MDRD level （MDRD〈90 or MDRD〉90）. Both SUA and creatinine were significantly higher in the lower MDRD group than those in the higher MDRD group （（301.90±96.46） vs. （264.07±84.74） μmol/L, and （89.10±31.00） vs. （66.37±11.15） μmol/L, respectively）. Logistic regression analysis showed that only age and SUA were the independent predictors of MDRD. Conclusion High-normal SUA was associated with albuminuria and impaired glomerular filtration rate in Chinese type 2 diabetic patients.

Etiology,treatment,and classification of Budd-Chiari syndrome

Since Professor WANG Zhong-gao＇s creative work on thesystemic treatment of the Budd-Chiari syndrome（BCS）, this debilitating disease has been more and more widely recognized in China. Several large-scale studies of surgery or intervention strategies for treating BCS have been reported. However, much controversy still remains regarding many aspects of this disease, including its etiology, treatment, and classification. This review explores these controversies with emphasis on areas that merit further study.

Effect of human epididymis protein 4 gene silencing on the malignant phenotype in ovarian cancer

Background Human epididymis secretory protein 4 （HE4） has been proved to be a promising novel biomarker for the detection of epithelial ovarian carcinomas. Compared with CA125, HE4 assay demonstrated an improved ability to discriminate between pelvic mass with malignant and benign disease. Though it is well known that HE4 is overexpressed in ovarian cancer, however, the role of HE4 in the carcinogenesis and progression of ovarian cancer remains unkown.Methods In this study, we explored the role of HE4 in the carcinogenesis and progression of ovarian cancer. We screened nine ovarian cancer cell lines for HE4 expression, and using RNA interference （RNAi）, we silenced HE4 gene expression in CaoV3 and SKOV3.ip1 ovarian cancer cell lines. We assessed the effect of HE4 gene silencing on the transformed phenotype by examining the cell cycle, apoptosis, proliferation and transwell migration/invasion in vitro.Results HE4 gene silencing induces G0/G1 arrest and blocks the progression from the G1 to S phase in CaoV3 and SKOV3.ip1 cells. HE4 knockdown also inhibited cell proliferation, migration and invasion in SKOV3.ip1 cells in vitro.Conclusion HE4 may be involved in the regulation of the cell cycle and promote ovarian cancer migration and invasion.

Radical surgery for Budd-Chiari syndrome through exposure of the entire inferior vena cava of the hepatic segment

Background Several kinds of radical surgery for the treatment of Budd-Chiari syndrome （BCS） have been devised. We have described preliminary efforts to treat BCS using a novel radical resection technique to expose the entire inferior vena cava （IVC） of the hepatic segment. Methods Sixty patients with BCS were treated by radical resection, including 46 men and 14 women. BCS patients ranged in age from 11 to 62 years, with 3 months to 11 years since the BCS diagnosis. The lesions included membrane occlusion of the IVC in 16 patients, double membranes within the IVC in 2 patients, double membranes within the IVC and the hepatic vein （HV） in 3 patients, IVC membrane with distal thrombosis in 10 patients, long segment thrombosis of the IVC in 5 patients （organized thrombosis in 2 patients, fresh thrombosis in 3 patients）, occlusion of the outlet of the HVs due to mural thrombosis in 2 patients, segmental occlusion of the IVC in 3 patients, membranes within the HV with IVC stenosis due to protrusion of HV stent in 1 patient, HV membranes in 11 patients, extensive occlusion of HVs in 1 patient, the whole IVC tumor thrombus with tumor thrombus of 2/3 right atrium resulting from a posterior peritoneum tumor in 1 patient, IVC leiomyosarcoma in 2 patients, IVC leiomyosarcoma with tumor thrombus into 1/2 right atrium in 1 patient, IVC thrombosis extending into right atrium in 1 patient, compression of supra-hepatic segment of IVC due to fiber trabs in 1 patient. Results All lesions were successfully resected under direct supervision. Three procedures were performed under extracorporeal circulation, 52 patients with catheterization of the right atrium, 4 patients with a cell saver, and one patient with auto-retrieval of blood. The retrieved blood was from 300 ml to 4000 ml. Transfusion of banked blood was from 400 ml to 2000 ml for 14 patients. For the other patients no transfusion of banked blood was required. One patient died of renal failure peri-operatively. Newly formed IVC membrane was found for one recurrent patient whose IVC thrombosis was removed one year prior. Restenosis of the IVC was observed post-operatively without symptoms in one patient. In the other patients, no recurrent symptom was found during the follow-up periods. Conclusion This novel surgery provides a clear visual field during the procedure and yields satisfactory short and Iona-term results.

Clinical effect of distal radius fracture treated with open reduction and internal plate fixation

Background For some specific comminuted unstable intra-articular fracture,the plaster cast can not maintain the alignment of the articular surface effectively.The aim of this study was to evaluate the clinical effects of distal radius fracture treated with open reduction and internal plate fixation retrospectively.Methods From January 2002 to March 2010,539 cases of distal radius fracture were treated with open reduction and internal fixation,including 184 males and 355 females aging 21-72 years （mean 57 years）.Fractures were caused by falling to the ground in 459 cases,by traffic accident in 62 cases and by athletic injuries in 18 cases.Of 539 cases,there were 523 cases of closed fracture and 16 cases of open fracture.According to Arbeitsgemeinschaft fur Osteosynthesefragen （AO） standards of classification,there were 14 cases of A2 type,22 of A3 type,18 of B1 type,24 of B2 type,62 of B3 type,91 of C1 type,162 of C2 type and 146 of C3 type.The time from injury to operation was 1-16 days （mean 5 days）.All patitents received open reduction and internal plate screw fixation.Forty-seven patients with bone defect were given 6-15 g autologous ilium and 75 cases were given 5 ml calcium sulphate artificial aggregate after reduction.Results All incisions healed by first intention after operation.Patients were followed up for 15 to 32 months postoperatively （mean 22 months）.The fractures healed within 10-18 weeks after operation （mean 12 weeks）.During the last follow-up,the mean palmar tilt was （7.0±0.9）° and the mean ulnar variance was （21.0±4.2）°,showing significant difference when compared with preoperation （（-5.0±1.2）° and （8.0±3.8）°）.The radial heights were not abbreviated.According to Gartland and Werley assessment system,the results were excellent in 314 cases,good in 163 cases,fair in 46 cases,and poor in 16 cases 12 weeks after operation,the excellent and good rate was 88.5%.Conclusions The clinical effect of distal radius fracture treated with open reduction and internal plate fixation was relatively satisfactory.Meticulous operation procedure and individual rehabilitation strategy contribute to the wrist joint functional recovery.

Erythrocytic or serum hydrogen sulfide association with hypertension development in untreated essential hypertension

Background Endogenous hydrogen sulfide （H2S） plays an important role in hypertension. The aim of this study was to investigate the role of erythrocyte and serum H2S in patients with untreated essential hypertension. Methods We recruited 62 patients （age 22-74 years） with untreated prehypertension or hypertension, and 64 normotensive subjects （age 18-64 years）. We assessed the 3-mercaptopyruvate sulphurtransferase （MPST） protein expression in erythrocytes and measured the H2S production from erythrocytes and serum H2S levels, then analyzed the association of erythrocytic or serum H2S content and blood pressure or cardiovascular risk factors （e.g., age, body mass index （BMI） and dyslipidemia）. A stepwise regression analysis was used to evaluate the possible relationship of erythrocytic H2S in hypertension. Results In hypertensive patients, erythrocyte H2S production （（111.04_＋.29.20） nmol/min per 10a erythrocytes） was higher than that in controls （（78.85＋19.38） nmol/min per 108 erythrocytes）, and serum H2S was also higher. The erythrocytic H2S production was associated with increased systolic blood pressure （sBP）, diastolic blood pressure （dBP）, age, BMI, level of C-reactive protein （CRP）, as well as triglycerides （TG） and high density lipoprotein cholesterol （HDL-C） Serum H2S was not associated with age or CRP. Stepwise regression analysis showed that erythrocytic H2S production was correlated with sBP, TG, HDL-C, low density lipoprotein cholesterol （LDL-C） and blood urea nitrogen （BUN） and serum H2S was correlated with dBP and TG. Results of receiver-operating characteristic curve analysis suggested that erythrocytic H2S production was a more sensitive predictor of hypertension development than serum H2S. Conclusion Ervthrocvtic or serum H^S Production is sensitive predictor of hvDertension.

IDENTIFICATION OF MYCOBACTERIUM MARINUM 65 KD HEAT SHOCK PROTEIN GENE BY POLYMERASE CHAIN REACTION RESTRICTION ANALYSIS FROM LESIONS OF SWIMMING POOL GRANULOMA

Background Nontuberculous mycobacterium （NTM） had been reported to cause cutaneous infections which are difficult to interpret due to the variability of the clinical manifestations. Among NTM infections, Mycobacterium marinum （M. marinum） are mostly seen to cause skin infection. It is therefore important to establish a rapid approach for detection and identification of M. marinum from lesions of patients with suspected M. marinum infections. Methods Specimens were obtained from 5 patients with swimming pool granuloma. DNA was extracted and polymerase chain reaction （PCR） was performed. PCR products were digested with Hae III and BstE II, then analysed by pattern restriction analysis to detect heat shock protein （hsp） 65 kD gene. Results The 65 kD hsp gene was found in all specimens from patients with swimming pool granuloma. PCR restriction analysis （PRA） identified all 5 samples to be M. marinum infections, and the result was consistent with that of routine bacteriological identification. The lesions subsided or markedly improved upon treatment. Conclusions PRA is a sensitive, specific and rapid method in identification of mycobacteria. Application of this method will be helpful for early diagnosis of mycobacterial skin infections.

The role of cyclooxygenase-2/prostanoid pathway in visceral pain induced liver stress response in rats

Background Cyclooxygenase （COX） is the rate-limiting enzyme in the production of prostanoids from arachidonic acid. COX-2 is the inducible enzyme in the COX family, together with the prostanoids forms the COX-2/prostanoid pathway. Research showed that the COX-2/prostanoid pathway is activated in hepatic diseases and liver stress reaction, such as fibrogenesis, portal hypertension, carcinogenesis, and ischemic/repeffusion injury. But there was no report on visceral pain induced liver stress. This study was to investigate the role of the COX-2/prostanoid pathway in liver stress response in rat acute colitis visceral pain liver stress model.Methods Fifty-three male SD rats were randomly divided into Naive, Model, NS398 treatment, and Morphine treatment groups. The rat acute colitis visceral pain liver stress model was established under anesthesia by the colonic administration of 0.5 ml of 6% acetic acid using a urethral catheter. NS398 and morphine were administrated 30 minutes prior to model establishment in NS398 and Morphine treatment groups respectively. Spontaneous activities and pain behavior were counted and the extent of colonic inflammation was assessed histologically. Liver tissue levels of Glutathione-S-Transferase （GST） activity, COX-2 mRNA, prostaglandin E2 （PGE2）, thromboxane B2 （TXB2） and 6-Ketone-prostaglandin F1α （6-K-PGF1α） contents were assessed.Results Thirty minutes after the colonic administration of acetic acid, a significant decrease in spontaneous activities and an increase in pain behaviors were observed in Model group （P〈0.01 and P〈0.05 respectively）, accompanied by colonic inflammation. Liver GST activity levels significantly dropped （P〈0.05）. Liver COX-2 mRNA expressi.on significantly increased, accompanied by an increase in liver concentrations of PGE2 and TXB2, but no obvious change in 6-K-PGF1α concentrations. NS398 and morphine both ameliorated post-stress liver GST activity （P〈0.05 and P〈0.01 respectively）, decreased stress-induced COX-2 expression, decreased PGE2 and TXB2 production, but increased liver 6-K-PGF1α levels. Morphine attenuation in colonic tissue inflammation was apparent at 24 hours （P〈0.05）.Conclusions Acute colitis visceral pain liver stress can induce liver injury. Liver injury might have occurred through the activation of the COX-2/prostanoid pathway and increased production of PGE2 and TXB2. Effective analgesia might offer protective effect during visceral pain stress.

Application of endobronchial ultrasound-guided transbronchial needle aspiration in the diagnosis of mediastinal lesions

Background Mediastinal lesions are often difficult to diagnose in clinical practice because of the unique anatomical position of the mediastinum, which makes performance of biopsy difficult. The value of endobronchial ultrasound-guided transbronchial needle aspiration in the diagnosis of lung cancer and mediastinal lymph node staging has been widely accepted. However, few studies have been conducted on the value of endobronchial ultrasound-guided transbronchial needle aspiration in the diagnosis and differential diagnosis of mediastinal lesions. The current study was conducted to investigate the value of endobronchial ultrasound-guided transbronchial needle aspiration in the diagnosis and differential diagnosis of isolated mediastinal lesions without lung abnormalities. Methods We retrospectively analyzed the data of patients with isolated mediastinal lesions without lung abnormalities for whom endobronchial ultrasound-guided transbronchial needle aspiration examination was performed at the Department of Thoracic Surgery of Peking University People＇s Hospital, between September 2009 and December 2010. For patients who could not be diagnosed with endobronchial ultrasound-guided transbronchial needle aspiration, surgical biopsy or more than 6 months of clinical and imaging follow-up was carried out. Results Endobronchial ultrasound-guided transbronchial needle aspiration was performed for 60 patients with isolated mediastinal lesions. Correct diagnosis was made in 48 cases. Nineteen cases were malignant, and 29 were benign. The rate of correct diagnosis was 80%. The sensitivity, specificity, and accuracy of endobronchial ultrasound-guided transbronchial needle aspiration in distinguishing benign from malignant mediastinal lesions were 95%, 100%, and 98%, respectively. The examination was tolerable for all patients. No associated complications were observed. Conclusion Endobronchial ultrasound-guided transbronchial needle aspiration is a safe and effective method of diagnosing mediastinal lesions.

Detecting PML-RARα transcript in acute promyelocytic leukemia using real-time quantitative RT-PCR

Background Real-time quantitative RT-PCR （RQ-PCR） assay has become a vital tool to monitor residual disease of leukemia, However, the complexity and standardization of RQ-PCR should never be overlooked and the results should be interpreted cautiously in clinical conditions. We aimed to assess the methodology of RQ-PCR and its clinical applications in monitoring molecular kinetics of 36 newly diagnosed cases of acute promyelocytic leukemia patients with t （15;17） from October 2004 to December 2005.Methods All the TaqMan probe-based RQ-PCR reactions and analysis were performed on an ABI-PRISM 7500 platform, The quantitation of PML-RARα transcripts was represented by the normalized quotient, that is, PML-RARα transcript copies divided by ABL transcript copies, According to induction therapy, the patients were classed into two groups： group 1 （n=23）, three-drug combination including arsenics, all-trans retinoic acid and mitoxantrone; and group 2 （n=13）.two-drug combination from all-trans retinoic acid, arsenics and mitoxantrone.Results The sensitivity of RQ-PCR was 1 per 105 cells and 5 copies of the PML-RARα transcript could be reproducibly detected, No false positive results occurred in 40 non-acute promyelocytic leukemia samples, Optimal amplification efficiency could be attained, which was determined by the slope of the standard curves （slope： -3.2 -- -3.7）. The inter-assay and intra-assay variation coefficients of the method were 1.01% and 0.56% respectively. Although the time to attain hematological complete remission was similar in both groups, the time to achieve molecular remission of group 1 was significantly shorter than that of group 2 （61 days vs 75 days, P=0.034）. The rate of molecular remission within 70 days was higher in group 1 than in group 2 （75.00% vs 38.46%, P=0.036）, Compared with pretreatment, median reduction of the PML-RARα transcript before first consolidation therapy differed significantly between group 1 and group 2 （log scale, 3.15 vs 2.31, P=0.024）, Interestingly, we found that PML-RARα transcript levels temporarily increased in bone marrow （7 patients） and peripheral blood （22 patients） samples of patients during induction therapy in both groups.Conclusions The RQ-PCR assay is reliable for the detection of PML-RARα transcripts. Arsenics, all-trans retinoic acid and mitoxantrone triad induction treatment of acute promyelocytic leukemia is superior to two-drug combination induction therapy in terms of the molecular response.

Modified conditioning regimen busulfan-cyclophosphamide followed by allogeneic stem cell transplantation in patients with multiple myeloma

Background Allogeneic stem cell transplantation is a potential curative approach in patients with multiple myeloma. The very high transplant related mortality associated with standard allogeneic stem cell transplantation is currently the major limitation to wider use of this potentially curative treatment modality. The challenge for clinical investigators is to reduce the incidence of post-transplant complications for patients receiving autologous hematopoietic stem cell transplantion for multiple myeloma. In this study the toxicity and efficacy of modified myeloablative conditioning regimen followed by allogeneic stem cell transplantation was investigated in patients with multiple myeloma. Methods The conditioning regimen consisted of hydroxyurea, cytarabine, busulfan, cyclophosphamide, and semustine. Ten patients underwent allogeneic transplantation among them hydroxyurea （40 mg/kg） was administered twice on day -10 and cytarabine （2 g/ms） was given on day -9, busulfan was administered orally in four divided doses daily for 3 days （days -8 to -6）. The dose of busulfan was 12 mg/kg in the protocol followed by cyclophosphamide intravenously over 1 hour on days -5 and -4 （1.8 g/m^2）, and with semustine （Me-CCNU） 250 mg/m^2 on day -3. Results Chimerism data were available on all patients and all patients achieved full donor chimerism without graft failure. Six patients had not acute graft-versus-host disease （GVHD, 36.4%; 95% CI： 13.9%-38.6%）. Two patients （18.2%） developed grade Ⅰ acute GVHD （95% CI： 10.9%-35.9%） and grade Ⅱ acute GVHD occurred in one patient （9.1%; 95% Cl. 8.4%-32.3%）. Severe grade IVa GVHD was seen in one patient, who died from acute GVHD. The incidence of chronic GVHD was 22.2% （95% Cl： 11.7%-36.7%）, among them one died of severe grade IV GVHD and one developed multiorgan failure on day ＋170; the treatment-related mortality was 22.0% （95% Cl： 10.3%-34.1%）. The overall 4-year survival rate was 67.8% （95% Cl： 16.3%-46.7%）. The estimated 4-year progression-free survival rate was 58.5% （95% CI： 13.7%-41.8%）. The 4-year complete remission was 72.7% （95% CI： 27.8%-49.6%）. One patient relapsed after 4 months and achived the complete remission after receiving the donor lymphocyte infusion. Conclusions Modified conditioning regimen busulfan-cyclophosphamide with peripheral blood stem cells＋bone marrow cells transplantation result in a low incidence of severe GVHD with a relatively low treatment-related mortality, high complete remission rates and a long-term survival.

Humoral immune responses induced by anti-idiotypic antibody fusion protein of 6B11scFv/hGM-CSF in BALB/c mice

Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B 11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody is COC166-9 （Abl）. In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment （scFv）/human granulocyte macrophage colony-stimulating factor （hGM-CSF） and 6B 1 lscFv in BALB/c mice. Methods The fusion protein 6B 11 scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSE BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. Results The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F（ab）2＇ and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6BllScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11 scFv/hGM-CSF compared with the 6B11 scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. Conclusion The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11 scFv/hGM-CSF for active immunotherapy of ovarian cancer.