[Clinical features and spinal lesions in patients with chronic prostatitis/chronic pelvic pain syndrome].

Objective: To analyze the clinical features and spinal lesions related to micturitionin of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS) patients. Methods: Patients with CP/CPPS were enrolled to this study at the outpatient department of Tongji Hospital between January and June 2019. The data of clinical features was collected and analyzed, including lower urinary tract symptoms(LUTS), bowel syndrome and pain over different parts of body, as well as lower urinary tract dysfunction, spinal lesions and pelvic organ morphological changes demonstrated by MRI. The potential role of spinal lesions in the development of CP/CPPS syndrome was investigated. Results: A total of 126 CP/CPPS patients were included, with an age［M（Q1，Q3）］of 41(31,53) years and a course of disease of 2(1,20) years. Among them, 126 (100.0%) were complicated with LUTS, 72(57.1%) with bowel dysfunction and 88(69.8%) with pain. MRI showed the cervical central disc herniation(126 cases, 100.0%), the ischemic changing in the cervical area of visceral efferant pathway(82 cases, 65.1%), the lumbar central disc herniation(65 cases, 51.6%), and the sacral nerve cysts(97 cases, 77.0%) are commonly seen. In addition, the morphological changes in the visceral organs containing smooth muscle were demonstrated, including thickened bladder wall(91 cases, 72.2%), distended seminal vesicles(70 cases, 55.6%) and distended sigmoid colon/rectum(59 cases, 46.8%). Conclusions: CP/CPPS patients were characterized by the co-existence of LUTS, bowel dysfunction and somatic pain in one individual. The presence of multi-organ symptoms, combined with the high prevalence of spinal lesions associated with micturition reflex, suggesting the potential role of the spinal lesions in the development of CP/CPPS.

[Analysis of gene mutation profile of adult soft tissue sarcomas using high-throughput sequencing technology].

Objective: To understand the genetic variation of soft tissue sarcomas, and to provide a scientific evidence for the individualized treatment. Methods: The somatic mutation and germline mutation of 45 adult soft tissue sarcomas had been detected by high-throughput sequencing technology, the clinical data were also analyzed. Results: A total of 88 gene mutations were detected in 45 samples, including 78 single nucleotide variation (SNV), 13 insertion/deletion (Indel) and 19 copy number variation (CNV). The most common mutant genes are TP53, CDKN2A, MDM2, CDK4, NF1 and PTEN. Among them, the mutation rates of TP53-MDM2/MDM4-CDKN2A pathway, CDKN2A/CDK4/RB1 pathway, and RAS/NF1/PTEN/PI3K pathway were more frequent (32/88, 36%). In terms of immunotherapy biomarkers among 10 samples, the median value of tumor mutation burden was 2.02 muts/Mb (0-4.24 muts/Mb), and all were microsatellite stable. Conclusions: This study analyzes the genetic variation of soft tissue sarcoma, and determines the high-frequency gene mutations and pathways, which may be the potential drug targets. This finding can provide scientific evidences for the personalized treatment of soft tissue sarcoma.

[Treatment and analysis of prognostic factors of stage â ¡ and â ¢ undifferentiated high-grade pleomorphic sarcoma in extremities and trunk].

Objective: To evaluate the efficacy and prognostic factors of comprehensive treatment of undifferentiated high grade pleomorphic sarcoma (UHGPS) in extremities and trunk, including surgery, radiotherapy and chemotherapy. Methods: A retrospective analysis and follow-up of 131 UHGPS cases with clinical stage Ⅱ or Ⅲ in extremities and trunk soft tissue was performed to analyze the prognostic factors. Survival data were collected through follow-up. The survival rate was calculated with life table method and Kaplan-Meier survival curves were drawn. Survival rate between the two groups was compared using Log rank test. The multivariate analysis was performed using Cox regression model. Results: The median survival time of 131 patients was 41.6 months. The 1-year, 3-year and 5-year survival rates were 95.0%, 82.0%, and 77.0%, respectively. The 5-year recurrence-free survival rate was 81.0%, and the 5-year metastasis-free survival rate was 72.0%. Univariate analysis showed that the tumor size, initial or recurrence, surgical margin, AJCC stage, and with/without standard treatment were associated with overall survival (all P<0.05). Stratification analysis according to the American Joint Committee of Cancer (AJCC) stage showed that 5-year survival rate of stage Ⅱ patients with radiotherapy was 100.0%, which was higher than that of patients without radiotherapy (79.6%) and the difference was statistically significant (P=0.010); but no statistical significance of radiotherapy for stage Ⅲ and chemotherapy for stage Ⅱ or Ⅲ patients (all P>0.05). The multivariate analysis showed surgical margin (HR=4.220, P=0.002), with/without standard treatment (HR=4.040, P=0.030) were independent risk factors associated with prognosis of UHGPS patients. Conclusions: For UHGPS with stage Ⅱ or stage Ⅲ in extremities and trunk soft tissue, patients with complete resection and standard treatment have improved prognosis. Therefore, standard treatment, including extensive resection for the first surgery, should be performed according to expert consensus in order to increase the long-term survival rate. Adjuvant radiotherapy should be performed for stage Ⅱ patients.

[Analysis of clinicopathological characteristics and prognostic factors of foot and ankle soft tissue and bone tumors].

Objective: To evaluate the clinicopathological characteristics of foot and ankle soft tissue and bone tumor, and to analyze the prognosis and the related factors of malignant tumors in this site. Methods: 74 patients with soft tissue and bone tumors of foot and ankle from January 2006 to February 2017 were retrospectively analyzed. The clinicopathological characteristics, the treatment and survival status of malignant tumors were followed up, and the clinical and therapeutic factors related to prognosis were analyzed. Results: Of the 74 patients, 34 were males and 40 were females. The male to female ratio was 1∶1.18; the age ranged from 12 to 64 years and the median age was 42 years. Tumors located in forefoot of 22 cases, 22 in midfoot, 10 in hind foot, 14 in ankle joint and 6 in multiple sites. 14 cases were bone tumors, including 7 benign and 7 malignant, and 60 cases were soft tissue tumors, including 14 benign and 46 malignant. The most common malignant soft tissue tumors were synovial sarcomas (13 cases), and the most common benign soft tissue tumors were hemangiomas (4 cases). 44 cases of malignant tumors underwent surgery were followed up, of which were 7 bone and 37 soft tissue malignant tumors. Limb salvage surgeries were performed in 33 cases and amputation in 11 cases. The median follow-up time was 69.8 months, and the median survival time was 40.7 months. The 1-year, 3-year and 5-year survival rate of soft tissue malignant tumors was 88.0%, 73.0%, and 63.0%, respectively. The 1-year, 3-year and 5-year survival rate of bone malignant tumors was 86.0%, 57.0% and 57.0%, respectively. Univariate analysis showed that the prognostic factors affecting 5-year survival rate were tumor size and adjuvant therapy (P<0.05). Patient's gender, age, tumor location, histological type and surgical procedure had no effect on overall survival(P>0.05). Multivariate analysis showed that tumor size was an independent prognostic factor (RR=7.262, P=0.005). Conclusions: Forefoot and midfoot are more common in foot and ankle soft tissue and bone tumors. Synovial sarcoma is the most common diagnosis in malignant soft tissue tumors, and hemangioma is the most common diagnosis in benign soft tissue tumors. The prognostic factor of malignant soft tissue and bone tumors in foot and ankle is tumor size. Patients with the tumor size of 5 cm or more have a worse prognosis.

[The role of preoperative (18)F-FDG PET-CT in lymphatic metastasis diagnosis of cutaneous malignant melanoma on extremities and trunk].

Objective: To evaluate the clinical value of preoperative (18)F-Fludeoxyglucose ((18)F-FDG PET-CT) in lymphatic metastasis diagnosis of cutaneous melanoma on extremities and trunk. Methods: 112 patients with cutaneous melanoma pathologically of extremities and trunk from January 2006 to December 2016, who received (18)F-FDG PET-CT examination preoperatively, were retrospectively reviewed. The correlations between the maximal diameters of lymph nodes, the maximal standard uptake value (SUV) and the diagnostic impression grades of PET-CT examination, and the final pathological diagnosis were analyzed. The correlations between Breslow thickness of primary lesions and the diagnostic impression of PET-CT examination were also analyzed. All the above were analyzed with Receiver Operating Characteristic (ROC) curve to get the cut-off value. Based on the final results of pathological diagnosis of lymph nodes as the golden standard, the statistically significant indicators of ROC curve analysis were used to evaluate the diagnostic effect, as well as to calculate the sensitivity, specificity and accuracy. With gender, age, maximal diameter of lymph nodes, maximal SUV, diagnosis impressions, and Breslow thickness as the independent variables and pathological diagnosis results of lymph nodes as the dependent variable, two-class stepwise Logistic regression analysis was used to determine the independence of diagnostic indicators. ROC curve analysis and log rank test were used to analyze the relationship between Breslow thickness and patient survival. Results: To evaluate melanoma patients' lymph node status, the results of ROC curve analysis showed that the area under the curve of lymph node maximal diameter, maximal SUV, diagnosis impression of PET-CT examinations were 0.789, 0.786 and 0.816, respectively (all P<0.05). The cut-off values were 0.85 cm, 1.45 and 2.5, respectively. The sensitivity of the cut-off values to determine the status of lymph nodes in melanoma patients were 71.4%, 64.9% and 72.1% respectively, and the specificities were 85.2%, 88.7% and 87.0% respectively. Multivariate Logistic regression analysis showed that PET-CT diagnosis impressions had independent diagnostic significance for the lymph node status of melanoma patients (OR=11.296, 95%CI: 2.550~50.033). The area under the curve of Breslow thickness evaluating PET-CT diagnostic impression is 0.664 (P=0.042) and the cut-off value was 4.25 mm. The survival rate of the patients with Breslow thickness ≥ 4.25 mm was lower than that in the group <4.25 mm (P=0.006). Conclusions: (18)F-FDG PET-CT can help to evaluate metastases and make treatment decisions for cutaneous melanoma of extremities and trunk, especially for patients whose primary lesion's Breslow thickness has reached more than 4.25 mm. For the patients whose maximal SUV of regional lymph node is higher than 1.45 and short diameter of the largest lymph node is larger than 0.85cm, the possibility of metastases should be considered.

Decreased expression of long non-coding RNA LINC00261 is a prognostic marker for patients with non-small cell lung cancer: a preliminary study.

OBJECTIVE: The previous study found that long non-coding RNA LINC00261 (LINC00261) was significantly down-regulated in non-small cell lung cancer (NSCLC). However, the function of LINC00261 in the progression of NSCLC has not been reported. The present work aimed to explore the prognostic value of LINC00261 in patients with NSCLC. PATIENTS AND METHODS: The expression of LINC00261 was determined in NSCLC tissues and matched normal lung tissues by quantitative Real-time PCR (qRT-PCR). Furthermore, we evaluated the relationship of LINC00261 and clinicopathological features with survival of patients with NSCLC. Finally, univariate and multivariate Cox regression analyses were used to explore whether LINC00261 was an independent predictor of survival. RESULTS: We found that the LINC00261 expression level in NSCLC tissues was suppressed compared with that in adjacent normal lung tissues (p < 0.01). Low expression of LINC00261 was found to significantly correlate with TNM stage (p = 0.005), lymph node status (p = 0.020), and distant metastasis (p = 0.004). Then, Kaplan-Meier analysis demonstrated that low LINC00261 expression level was associated with poorer overall survival (p = 0.0013). Furthermore, multivariate analysis showed that low expression of LINC00261 was an independent adverse prognostic factor of NSCLC (p = 0.004). CONCLUSIONS: We firstly provided evidence that LINC00261 expression was associated with poor prognosis of NSCLC patients and may serve as an independent prognostic indicator.

[Clinicopathological features and prognosis of malignant peripheral nerve sheath tumor: a retrospective study of 140 cases].

Objective: To investigate the clinicopathological features and prognosis of malignant peripheral nerve sheath tumors (MPNST). Methods: We retrospectively reviewed the clinical data of MPNST patients who were treated at Cancer Institute & Hospital, Chinese Academy of Medical Science from January 1999 to January 2016. A total of 140 patients with 66 male and 74 female with MPNST were enrolled in the study. The median age was 40 at the time of diagnosis. Survival analysis were estimated by Kaplan-Meier method and Log rank test. Multivariate analysis were estimated by Cox proportional hazards regression model. Results: The median follow-up time was 43.0 months. The 3- and 5-year overall survival (OS) rates were 56.4% and 48.6%, respectively. The 3-year local recurrence (LR) rate and distant metastasis (DM) rates were 42.9% and 49.3%, respectively. Univariate analysis showed that the tumor location, AJCC stage, S-100, radiotherapy and margin status affected 5-year OS rate (all P<0.05). The tumor location, AJCC stage, S-100, Ki-67 staining, margin status, radiotherapy and chemotherapy affected 3-year LR rate (all P<0.05). The tumor location, AJCC stage, S-100, Ki-67 staining and margin status affected 3-year DM rate (all P<0.05). Multivariate analysis showed that the tumor location, AJCC stage, S-100 were independent factors for 5-year OS rate (all P<0.05). The tumor location, Ki-67 staining and chemotherapy were independent factors for LR (all P<0.05) while the AJCC stage, margin status and Ki-67 staining were independent factors for DM (all P<0.05). Conclusions: MPSNT is an aggressive tumor with poor prognosis. Multiple factors were identified in this study. Patients with the tumor located at head and neck, advanced AJCC stage and negative S-100 usually have a low 5-year overall survival rate. Patients with the tumor located at head and neck, Ki-67 staining ≥ 20% and without chemotherapy had a higher tendency of local recurrence. Poor prognosis factors for DM were advanced AJCC stage, positive margin and Ki-67 staining ≥ 20%.

Comparison of mercury sulfides with mercury chloride and methylmercury on hepatic P450, phase-2 and transporter gene expression in mice.

Zuotai (mainly ß-HgS) and Zhusha (also called as cinnabar, mainly α-HgS) are used in traditional medicines in combination with herbs or even drugs in the treatment of various disorders, while mercury chloride (HgCl2) and methylmercury (MeHg) do not have known medical values but are highly toxic. This study aimed to compare the effects of mercury sulfides with HgCl2 and MeHg on hepatic drug processing gene expression. Mice were orally administrated with Zuotai (ß-HgS, 30mg/kg), α-HgS (HgS, 30mg/kg), HgCl2 (33.6mg/kg), or MeHg (3.1mg/kg) for 7days, and the expression of genes related to phase-1 drug metabolism (P450), phase-2 conjugation, and phase-3 (transporters) genes were examined. The mercurials at the dose and duration used in the study did not have significant effects on the expression of cytochrome P450 1-4 family genes and the corresponding nuclear receptors, except for a slight increase in PPARα and Cyp4a10 by HgCl2. The expressions of UDP-glucuronosyltransferase and sulfotransferase were increased by HgCl2 and MeHg, but not by Zuotai and HgS. HgCl2 decreased the expression of organic anion transporter (Oatp1a1), but increased Oatp1a4. Both HgCl2 and MeHg increased the expression of multidrug resistance-associated protein genes (Mrp1, Mrp2, Mrp3, and Mrp4). Zuotai and HgS had little effects on these transporter genes. In conclusion, Zuotai and HgS are different from HgCl2 and MeHg in hepatic drug processing gene expression; suggesting that chemical forms of mercury not only affect their disposition and toxicity, but also affect their effects on the expression of hepatic drug processing genes.

Correlation between polymorphism of platelet alloantigen genes HPA-1-5 and type 2 diabetes complication by carotid atherosclerosis in a Chinese population.

We investigated the association between the polymorphism of human platelet alloantigen genes HPA-1-HPA-5 and the complication of type 2 diabetes mellitus (T2DM) by carotid atherosclerosis (CA) among Han people in Guiyang District, China. Ninety-nine T2DM patients were selected from the Affiliated Hospital of Guiyang Medical College and divided into a CA(+) group and a CA(-) group. A control group comprised 100 healthy people from the medical examination center of the same hospital. Genomic DNA from all the subjects was isolated by phenol-chloroform extraction and target genes were amplified using sequence-specific primer-polymerase chain reaction, followed by gene type detection of HPA. There were significant differences in allele and genotype frequencies of HPA-1, -2, -3, and -5 among the three groups [CA(+), CA(-), and the control group] (P < 0.05), and significant differences in allele and genotype frequencies of HPA-1, -2, and -3 between groups CA(+) and CA(-) and the control group (P < 0.05). Moreover, there was a significant difference in allele and genotype frequencies of HPA-5 between the CA(+) and CA(-) groups (P < 0.05). Logistic regression analysis showed that risk factors for T2DM patients developing a CA complication were age, duration of diabetes, high blood pressure, smoking, overweight, abnormal blood lipid levels, and polymorphism of HPA-5. There may be a correlation between T2DM and polymorphism of HPA-1-3. Polymorphism of HPA- 5 is probably a risk factor for CA complicating T2DM.

Clinical analysis of osteosarcoma patients treated with high-dose methotrexate-free neoadjuvant chemotherapy.

OBJECTIVE: High-dose methotrexate (hdmtx) is a common therapeutic agent in the treatment of osteosarcoma. However, hdmtx is highly toxic and requires complex pharmacokinetic monitoring and leucovorin rescue. Thus, alternative therapeutic strategies are necessary. Here, we analyzed the clinical efficacy of a dia regimen (cisplatin-ifosfamide-doxorubicin) to evaluate its potential as an alternative to hdmtx-based therapy. METHODS: Patients received 12 cycles of chemotherapy administered over 2 years (2 preoperative cycles and 10 postoperative cycles). Cumulative dose was the same in all cycles: cisplatin 120 mg/m(2) on day 1 of week 1, followed by ifosfamide 2.0 g/m(2) days 1-5 of week 2, and doxorubicin 20 mg/m(2) days 1-3 of week 2. RESULTS: Between January 2004 and October 2008, 39 eligible patients (median age: 16 years) were enrolled, with 36 being evaluable for the study. Of those 36 patients, 20 (55.6%) had a good histologic response to preoperative chemotherapy (>90% tumour necrosis). The estimated 5-year rates of event-free survival (efs) and overall survival were 54.8% and 61.5% respectively. CONCLUSIONS: The results of our study suggest that, in osteosarcoma patients, the dia regimen produces an efs rate and survival outcomes comparable to those attained with hdmtx-containing regimens, with fewer adverse reactions. The dia regimen is well tolerated, and we observed a high level of patient compliance. Our results demonstrate that hdmtx-free osteosarcoma treatment regimens can be effective, warranting further investigation.

Successful management of a childhood osteosarcoma with epiphysiolysis and distraction osteogenesis.

In an 11-year-old boy with osteosarcoma in the proximal tibia (type iii), 2 cycles of dia chemotherapy (cisplatin, ifosfamide, doxorubicin) were administered preoperatively while epiphysiolysis was performed. Clinical response was determined to be complete by radiography and histopathology. Marginal excision was then performed with preservation of the proximal tibial epiphysis. Metaphyseal reconstruction was performed using distraction osteogenesis. Six cycles of dia chemotherapy were administered postoperatively. Twenty months later, the patient had developed no complications and experienced full bone healing, with no limb discrepancy. In selected adolescent patients with osteosarcoma, in whom the tumour is in full contact with the epiphysis, epiphyseal preservation by epiphysiolysis and reconstruction by distraction osteogenesis can provide an excellent outcome, resulting in a stable reconstruction that functionally restores the native limb.

Composite artificial semi-knee joint system.

OBJECTIVES: The purpose of the study was to investigate the clinical implantation protocol of custom-made artificial semi-knee joint based on computer-aided design so as to improve the limb salvage efficiency. MATERIALS AND METHODS: The custom-made artificial semi-knee joint was designed and manufactured based on rapid prototyping technology. The repeated modifications were carried out in the design and manufacture of the semi-knee joint, together with the operation protocol. Clinical trial was conducted on 2 cases of osteosarcoma, one receiving allograft prosthesis composite transplantation, and the other receiving synthetic bone graft prosthesis composite transplantation. The clinical outcomes of the 2 patients were evaluated. RESULTS: The custom-made artificial semi-knee joint met the clinical customization needs. In clinical trial, 18-month follow-up demonstrated the satisfactory knee joint function recovery in near future. CONCLUSIONS: The custom-made artificial semi-knee joint based on computer-aided design can afford satisfactory knee joint function recovery following allograft bone transplantation.

Effects of polymorphisms in the XRCC1, XRCC3, and XPG genes on clinical outcomes of platinum-based chemotherapy for treatment of non-small cell lung cancer.

This study aimed to investigate the effects of single-nucleotide polymorphisms (SNPs) XRCC1 Arg194Trp, XRCC1 Arg280His, XRCC1 Arg399Gln, XRCC3 Thr241Met, XPG His104Asp, and XPG His46His in genes involved in the DNA-repair pathway on the outcomes of platinum-based chemotherapy in patients with advanced non-small cell lung cancer (NSCLC). The study period was from January 2005 to January 2006, and 378 NSCLC patients were enrolled within 1 month after being diagnosed with NSCLC. Genomic DNA was extracted using the Qiagen Blood Kit. Polymerase chain reaction combined with a restriction fragment length polymorphism assay was used for genotyping. Individuals with the XRCC1 399A/A genotype had a higher probability of responding well to platinum-based chemotherapy, indicated by an odds ratio (OR) of 2.27 [95% confidence interval (CI)=1.64-6.97]. Similarly, the XPG T/T genotype was significantly associated with improved responses to chemotherapy, indicated by an OR of 1.90 (95%CI=1.10-3.28). The XRCC1 399A/A genotype was significantly associated with longer disease-free survival and overall survival, indicated by hazard ratios (HRs) of 0.48 (95%CI=0.25-0.88) and 0.51 (95%CI=0.26- 0.98), respectively. Moreover, the XPG 46T/T genotype increased the likelihood of longer disease-free survival and overall survival of NSCLC patients treated with platinum-based chemotherapy (HR=0.47; 95%CI=0.22-0.82 and HR=0.52; 95%CI=0.31- 0.96, respectively). These results indicate that XRCC1 Arg399Gln and XPG His46His might significantly affect the clinical outcomes of platinum-based chemotherapy, highlighting the need for larger studies to confirm the role of these two SNPs in outcomes of NSCLC treatments.

Denosumab and giant cell tumour of bone-a review and future management considerations.

Giant cell tumour of bone (gctb) is one type of giant-cell-rich bone lesion characterized by the presence of numerous multinucleated osteoclast-type giant cells. Giant cells are known to express rankl (receptor activator of nuclear factor κB ligand) and are responsible for the aggressive osteolytic nature of the tumour. No available treatment option is definitively effective in curing this disease, especially in surgically unsalvageable cases. In recent years, several studies of denosumab in patients with advanced or unresectable gctb have shown objective changes in tumour composition, reduced bony destruction, and clinical benefit. Denosumab is a fully human monoclonal antibody that targets and binds with high affinity and specificity to rankl. Several large phase iii studies have shown that denosumab is more effective than bisphosphonates in reducing skeletal morbidity arising from a wide range of tumours and that it can delay bone metastasis. The relevant articles are reviewed here. The controversies related to the future use of denosumab in the treatment of gctb are discussed.

Analysis of HLA-A, HLA-B and HLA-DRB1 alleles in Chinese patients with lung cancer.

The primary function of the human leukocyte antigen (HLA) system is to regulate the immune response. Because of its important role in the immune response and its high degree of polymorphism, the HLA system is associated with many diseases. We examined the polymorphisms of HLA-A, B and DRB1 alleles in 100 unrelated patients with lung carcinoma and in 438 unrelated normal controls of Han nationality from North China, using sequence-based typing and PCR with sequence-specific primers. We found that the frequencies of HLA-A*0201, A*2601, B*1518, B*3802, DRB1*0401, DRB1*0402, and DRB1*1201 were higher in the lung carcinoma group than in the normal control group. The P values were 0.035, 0.040, 0.001, 0.017, 0.014, 0.004, and 0.019, respectively, and the odds ratio values were 1.052, 3.513, 4.047, 3.054, 4.237, 19.397, and 2.128, respectively. The frequency of HLA-DRB1*1302 was lower in the lung carcinoma group than in the normal control group (P = 0.046, odds ratio = 0.168). We concluded that patients with lung cancer and healthy controls of Han nationality from North China differ in the frequencies of various HLA alleles.

Relationship between genetic polymorphism of MCP-1 and non-small-cell lung cancer in the Han nationality of North China.

Monocyte chemoattractant protein 1 (MCP-1) is an important chemokine that has a dose-dependent anti-tumoral effect. Polymorphism in the MCP-1 distal regulatory region (-2518A/G) can affect the level of MCP-1 expression. We examined the polymorphisms of 112 unrelated patients with non-small-cell lung cancer (NSCLC) and 82 unrelated healthy controls of Han nationality in North China using PCR-RFLP. We found that the distributions of AA, AG and GG genotypes of MCP-1-2518 were significantly different in NSCLC patients compared to controls (chi(2) = 10.106, P = 0.006). There was a significant increase in the frequency of the AA genotype (odds ratio (OR) = 3.138, chi(2) = 8.905, P = 0.003) and a significant decrease in the frequency of the GG genotype (OR = 0.516, chi(2) = 4.613, P = 0.032) in the NSCLC patients, compared to controls. The frequencies of AA, AG and GG genotypes did not differ in the NSCLC patients according to the number of pack-years smoked. Based on these results, we suggest that the MCP-1 -2518A/G polymorphism is associated with genetic susceptibility to NSCLC.

Distinct leukemia phenotypes in transgenic mice and different corepressor interactions generated by promyelocytic leukemia variant fusion genes PLZF-RARalpha and NPM-RARalpha.

Acute promyelocytic leukemia (APL) is characterized by a specific chromosome translocation involving RARalpha and one of four fusion partners: PML, PLZF, NPM, and NuMA genes. To study the leukemogenic potential of the fusion genes in vivo, we generated transgenic mice with PLZF-RARalpha and NPM-RARalpha. PLZF-RARalpha transgenic animals developed chronic myeloid leukemia-like phenotypes at an early stage of life (within 3 months in five of six mice), whereas three NPM-RARalpha transgenic mice showed a spectrum of phenotypes from typical APL to chronic myeloid leukemia relatively late in life (from 12 to 15 months). In contrast to bone marrow cells from PLZF-RARalpha transgenic mice, those from NPM-RARalpha transgenic mice could be induced to differentiate by all-trans-retinoic acid (ATRA). We also studied RARE binding properties and interactions between nuclear corepressor SMRT and various fusion proteins in response to ATRA. Dissociation of SMRT from different receptors was observed at ATRA concentrations of 0.01 microM, 0.1 microM, and 1.0 microM for RARalpha-RXRalpha, NPM-RARalpha, and PML-RARalpha, respectively, but not observed for PLZF-RARalpha even in the presence of 10 microM ATRA. We also determined the expression of the tissue factor gene in transgenic mice, which was detected only in bone marrow cells of mice expressing the fusion genes. These data clearly establish the leukemogenic role of PLZF-RARalpha and NPM-RARalpha and the importance of fusion receptor/corepressor interactions in the pathogenesis as well as in determining different clinical phenotypes of APL.

Autologous bone marrow mixed with HLA-haploidentical allogeneic marrow transplantation for treatment of patients with malignant blood diseases.

We have previously demonstrated that syngeneic marrow mixed with H-2 haploidentical marrow transplantation could provide not only protection against graft-versus-host disease (GVHD) but also anti-leukemic (GVL) effects in mice. In the present studies, we report clinical observations using autologous marrow mixed with HLA-haploidentical allogeneic marrow transplantation for treatment of patients with malignant blood diseases. Sixteen cases, including 12 with acute leukemia and four with advanced malignant lymphoma, were treated by autologous marrow, which was purged in vitro by hyperthemia (42.5 degrees C for 70 min) following incubation for 5 days with interleukin 2 (IL-2) in liquid culture and mixed with HLA haploidentical marrow cells from their sibling or parent. Acute GVHD was not observed in any patient after transplantation. Hematological rescue in the clinical setting was demonstrated in all cases but one who died early from hepatic veno-occlusive disease (VOD). Five cases who were transplanted at the time of CR2 or CR3 and in advanced phase of lymphoma, relapsed 4 to 7 months after transplantation. The relapse rate was 31.3%. None of eight patients who received allogeneic BMT within 2 h after ABMT relapsed with median follow-up of 12 months and two of them died from procedure-related complications. Seven cases are still alive and disease-free with a median follow-up of 12 months. Mixed chimerism was found in 3/6 cases, who had different sex donors, by analysis of sex chromosomes. These results show that mixed transplantation is a safe, effective and new approach to treating patients with malignant tumors. In order to detect the effects of GVL, studies are now in progress in our clinic.

Central expression of c-fos protein after peripheral noxious thermal stimulation in awake rats.

This study applied immunohistochemistry method to examine the pattern of c-fos expression in the neuraxis following peripheral noxious thermal stimulation accomplished by immersion of tail of awake rat into hot water (50 degrees C). In unstimulated control rats, no obvious baseline expression of c-fos protein was found except in nucleus paraventricularis hypothalami and colliculus inferior, probably associated with restraint-induced stress and auditory stimulus, respectively. Noxious thermal stimulation resulted in the activation of c-fos expression, and bilateral increased nuclear immunostaining was counted in dorsal horn of lumbar and sacral segments of spinal cord (laminae I, II), nucleus raphe dorsalis, substantia grisea centralis (ventralis), nucleus paraventricularis thalami, nucleus anterior thalami, nucleus ventralis thalami, nucleus medialis thalami, nucleus reuniens, nucleus rhomboideus, nucleus habenulae lateralis, nucleus paraventricularis hypothalami, nucleus arcuatus, nucleus lateralis hypothalami, nucleus preopticus lateralis, nucleus septi lateralis, nucleus amygdala, nucleus striae terminalis, nucleus tractus diagonalis, and cortex cerebri. The results demonstrated that peripheral noxious stimulation induced central c-fos protein expression in a pattern of labeling nociresponsive cells.