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INTRODUCTION: Breast cancer is a worldwide health problem and the leading cause of cancer death among females. We previously identified Jumonji domain containing 2A (JMJD2A) as a critical mediator of breast cancer proliferation, migration and invasion. We now report that JMJD2A could promote breast cancer progression through transcriptional repression of the tumor suppressor aplasia Ras homolog member I (ARHI). METHODS: Immunohistochemistry was performed to examine protein expressions in 155 cases of breast cancer and 30 non-neoplastic tissues. Spearman correlation analysis was used to analyze the correlation between JMJD2A expression and clinical parameters as well as several tumor regulators in 155 cases of breast cancer. Gene and protein expressions were monitored by quantitative polymerase chain reaction (qPCR) and Western blot. Results from knockdown of JMJD2A, overexpression of JMJD2A, Co-immunoprecipitation (Co-IP) assay, dual luciferase reporter gene assay and chromatin immunoprecipitation (ChIP) elucidated molecular mechanisms of JMJD2A action in breast cancer progression. Furthermore, the effects of ARHI overexpression on JMJD2A-mediated tumor progression were investigated in vitro and in vivo. For in vitro experiments, cell proliferation, wound-healing, migration and invasion were monitored by cell counting, scratch and Boyden Chamber assays. For in vivo experiments, control cells and cells stably expressing JMJD2A alone or together with ARHI were inoculated into mammary fat pads of mice. Tumor volume, tumor weight and metastatic nodules were measured by caliper, electronic balance and nodule counting, respectively. RESULTS: JMJD2A was highly expressed in human breast cancers and positively correlated with tumor progression. Knockdown of JMJD2A increased ARHI expression whereas overexpression of JMJD2A decreased ARHI expression at both protein and mRNA levels. Furthermore, E2Fs and histone deacetylases were involved in the transcriptional repression of ARHI expression by JMJD2A. And the aggressive behavior of JMJD2A in breast cancers could be reversed by re-expression of ARHI in vitro and in vivo. CONCLUSION: We demonstrated a cancer-promoting effect of JMJD2A and defined a novel molecular pathway contributing to JMJD2A-mediated breast cancer progression.
Assuntos
Neoplasias da Mama/genética , Histona Desmetilases com o Domínio Jumonji/genética , Transcrição Gênica/genética , Proteínas rho de Ligação ao GTP/biossíntese , Animais , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Fatores de Transcrição E2F/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , Células HEK293 , Histona Desacetilases/genética , Humanos , Histona Desmetilases com o Domínio Jumonji/biossíntese , Células MCF-7 , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica/genética , Transplante de Neoplasias , Regiões Promotoras Genéticas/genética , Ligação Proteica/genética , Interferência de RNA , RNA Mensageiro/biossíntese , RNA Interferente Pequeno , Transplante Heterólogo , Cicatrização/genética , Proteínas rho de Ligação ao GTP/genéticaRESUMO
Anaphylaxis is a rare but well-known cause of sudden unexpected death, although data from forensic autopsies in anaphylactic deaths are limited. Herein, a retrospective study of a series of allergic deaths from 2009 through 2019 in Shanghai, China, was conducted to investigate the demographic, medical, and forensic pathological characteristics of fatal anaphylaxis to improve medicolegal understanding on anaphylactic death. Sixty-two autopsy cases of anaphylactic death were registered in this study. Males dominated the cases (74.2%) against females (25.8%), with an average age of 38.8 years. Medications (98.4%), particularly antibiotics (72.6%), were the most frequent cause of anaphylaxis, and 44 cases (71.0%) occurred in clinics administered illegally by unlicensed clinicians. The anaphylactic symptoms began within a few minutes to less than 1 h in 53 cases, with dyspnea (56.5%) and sudden shock (46.8%) being the most common clinical signs. Thirty cases (48.4%) of anaphylaxis resulted in death within 1 h. Laryngeal edema and multiple tissue eosinophil infiltration (85.5%) were the most prevalent autopsy findings, followed by pulmonary edema and congestion (24.2%), which were considered to be non-specific but suggestive. The comorbidities were mainly cardiovascular disease (33.9%), pneumonia (8.1%) and asthma (8.1%). Serum IgE were measured in 11 of 62 cases, ranging from 43.3 to 591 IU/ml, severed as a helpful marker. Therefore, we suggested a thorough analysis of allergen exposure, clinical history and autopsy findings is required for the diagnosis of anaphylactic death currently.
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Jumonji Domain Containing 2A (JMJD2A) may be a cancer-associated gene involved in human breast cancer. With a view to investigating expression of JMJD2A in human breast cancer and benign lesion tissues as well as relationship between JMJD2A and tumor related proteins, histological and immunohistochemical analysis, Western blot and quantitative real-time PCR in infiltrating duct carcinoma and fibroadenoma for JMJD2A and immunohistochemical analysis and quantitative real-time PCR in infiltrating duct carcinoma for tumor related proteins (ARHI, p53, ER, PR and CerbB-2) were performed. Histological examination validated the clinical diagnosis. The JMJD2A positive rate of infiltrating duct carcinoma was significantly higher than fibroadenoma by immunohistochemical analysis. The mean optical density of JMJD2A in infiltrating duct carcinoma was higher than fibroadenoma by western blot. JMJD2A mRNA level in infiltrating duct carcinoma was higher than fibroadenoma by quantitative real-time PCR. Spearman correlation analysis revealed that the expression of JMJD2A was associated with ARHI, p53 and ER from immunohistochemical results respectively. Pearson correlation analysis revealed that the expression of JMJD2A was associated with ARHI, p53 and ER from quantitative real-time PCR results respectively. Expression of JMJD2A in infiltrating duct carcinoma was higher, and associated with ARHI, p53 and ER. The results may take JMJD2A as a potential diagnostic and therapeutic target in human breast cancer.
Assuntos
Neoplasias da Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Fibroadenoma/metabolismo , Histona Desmetilases com o Domínio Jumonji/biossíntese , Receptores de Estrogênio/biossíntese , Proteína Supressora de Tumor p53/biossíntese , Proteínas rho de Ligação ao GTP/biossíntese , Linhagem Celular Tumoral , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Receptor ErbB-2/biossíntese , Receptores de Progesterona/biossínteseRESUMO
Precisely determining the postmortem interval (PMI), which is crucial to criminal and forensic cases, is a research in which quantitative RT-PCR (also known as qRT-PCR or real-time RT-PCR) has been used to analyse gene expression levels and data normalisation should be required to eliminate the differences among the samples. Therefore, it is quite necessary to find stable molecular biological markers in PMI determination research. In this study, we compared nine commonly used endogenous markers (containing ACTB, GAPDH, B2M, U6, 18S rRNA, hsa-mir-1, hsa-mir-9, hsa-mir-194-1 and hsa-mir-203) in the 109 human tissue samples obtained from autopsy at the aim of finding stable markers in human tissues with consideration of the impact of parameters (PMI and cause of death). After RNA was extracted from four tissues (heart, brain, kidney, skin), the Ct values of nine endogenous markers were obtained by qRT-PCR and assessed by geNorm software. The results showed that U6, GAPDH and 18S rRNA were the suitable markers in our set of samples in various corpse conditions, that B2M and ACTB were reliable internal controls in heart tissue only, and that microRNAs had such high M values that they should not be chosen for endogenous control genes.
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Encéfalo/metabolismo , Expressão Gênica , Rim/metabolismo , Miocárdio/metabolismo , Mudanças Depois da Morte , Pele/metabolismo , Encéfalo/patologia , Genética Forense , Marcadores Genéticos , Humanos , Rim/patologia , Miocárdio/patologia , Reação em Cadeia da Polimerase em Tempo Real , Pele/patologiaRESUMO
microRNA (miRNA or miR) is a small single stranded non-coding RNA (21-25nt) that regulates gene expression in almost creatures. Currently, plenty of researches on how miRNA affects human cardiovascular disease have been reported. This review highlights recent findings about the role of miRNA in heart tissue and circulation correlated with human cardiovascular disease and explores the application of miRNA in sudden cardiac death in forensic science.
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Doenças Cardiovasculares/metabolismo , Morte Súbita Cardíaca/patologia , MicroRNAs/metabolismo , Embolia Pulmonar/diagnóstico , Animais , Biomarcadores/sangue , Cardiomiopatia Dilatada/sangue , Cardiomiopatia Dilatada/genética , Cardiomiopatia Dilatada/metabolismo , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/genética , Causas de Morte , Ciências Forenses/métodos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/genética , Insuficiência Cardíaca/metabolismo , Humanos , MicroRNAs/sangue , MicroRNAs/genética , Miocárdio/metabolismo , Miocárdio/patologia , Embolia Pulmonar/sangue , Regulação para CimaRESUMO
OBJECTIVE: To compare the effects of various kinesio tape applications on static balance and muscle activity in children with developmental coordination disorder. Methods: Four taping conditions were applied to 48 children with developmental coordination disorder: no taping, gastrocnemius taping, tibialis anterior taping; and peroneus longus taping. Postural sway and electromyographic data were assessed, with eyes closed (30 s), standing still in 2-leg stance, dominant-leg stance, and non-dominant-leg stance. RESULTS: Kinesio taping significantly reduced postural sway in both anteroposterior and mediolateral directions for dominant-leg stance and non-dominant-leg stance, but not 2-leg stance. During single-leg stances, anteroposterior sway was significantly lower for the gastrocnemius taping condition than for the no taping, tibialis anterior and peroneus longus taping conditions, and significantly lower in the tibialis anterior and peroneus longus taping conditions than in the no taping condition (gastrocnemius < tibialis anterior = peroneus longus < no taping). In addition, mediolateral sway was significantly lower in the tibialis anterior and peroneus longus taping conditions than in the no taping and gastrocnemius taping conditions, and significantly lower in the gastrocnemius taping condition than in the no taping condition (tibialis anterior = peroneus longus < gastrocnemius < no taping). Electromyographic data showed that muscle activity was significantly greater only for muscles where kinesio tape was applied. CONCLUSION: Various kinesio tape applications can differentially reduce postural sway and increase muscle activity during single-leg stances in children with developmental coordination disorder.
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Fita Atlética , Transtornos das Habilidades Motoras , Humanos , Criança , Músculos , Perna (Membro)RESUMO
Use of newer antipsychotics for substitution of current antipsychotics might be one way awaiting to be clinically verified to address antipsychotic cardiotoxic effects. Alternatively, the combination of existing antipsychotics with cardioprotective agents is also beneficial for patients with mental disorders for avoiding cardiotoxicity to the maximum.
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Chronic use of antipsychotic medications entails a dilemma between the benefit of alleviating psychotic symptoms and the risk of troubling, sometimes life-shortening adverse effects. Antipsychotic-induced cardiotoxicity is one of the most life-threatening adverse effects that raises widespread concerns. These cardiotoxic effects range from arrhythmia to heart failure in the clinic, with myocarditis/cardiomyopathy, ischemic injuries, and unexplained cardiac lesions as the pathological bases. Multiple mechanisms have been proposed to underlie antipsychotic cardiotoxicity. This review aims to summarize the clinical signs and pathological changes of antipsychotic cardiotoxicity and introduce recent progress in understanding the underlying mechanisms at both the subcellular organelle level and the molecular level. We also provide an up-to-date perspective on future clinical monitoring and therapeutic strategies for antipsychotic cardiotoxicity. We propose that third-generation antipsychotics or drug adjuvant therapy, such as cannabinoid receptor modulators that confer dual benefits - i.e., alleviating cardiotoxicity and improving metabolic disorders - deserve further clinical evaluation and marketing.
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The aim of this study was to determine the effects of different attentional focus conditions (external focus (EF) vs. internal focus (IF) vs. no focus (NF)) on the performance of a pole-holding task and postural stability in children with developmental coordination disorder (DCD) and typically developing children (TDC), while addressing previous study limitations. A total of 186 participants were enrolled in the analysis comprising 91 children with DCD (28 in the EF, 32 in the IF, and 31 in the NF condition) and 95 TDC (31 in the EF, 33 in the IF, and 31 in the NF condition). Participants were required to hold a pole with their hands still and parallel to the floor while focusing on pole movement (EF), focusing on hands (IF), or without focus instructions (NF), while pole movements and postural sway were recorded. Results showed that pole movements were significantly smaller during the EF condition (p < 0.05) compared with IF and NF conditions for both the DCD and TDC groups. In addition, postural sway was significantly lower during the EF condition compared with IF and NF conditions (p < 0.05). This study verified that the EF condition can not only facilitate better performance in holding a pole horizontal and immobile, but also concurrently promote greater postural stability than both IF and NF conditions for children with DCD and TDC.
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Transtornos das Habilidades Motoras/psicologia , Equilíbrio Postural , Atenção , Criança , Humanos , Desempenho PsicomotorRESUMO
OBJECTIVES: To compare sensitivity to light touch in children with developmental coordination disorder and those with typical development. Also, to investigate how changes/increases in sensitivity to light touch influence the effects of light fingertip touch on reducing body sway in both groups, while controlling for the confounding effects of arm configuration. METHODS: Twenty-six children with developmental coordination disorder and 26 typically developing children were enrolled in the study. To change/increase sensitivity to light touch, participants immersed their dominant index finger in a surfactant-water solution. Sensitivity to light touch was measured before and after soaking. Participants performed all conditions (no fingertip touch, light fingertip touch, and light fingertip touch after soaking) with the same arm configuration, while body sway was measured. RESULTS: Analysis of variance (ANOVA) revealed that the children with developmental coordination disorder were less sensitive to light touch than typically developing children (p <0.05). For both groups, immersing a fingertip in surfactant-water solution increased sensitivity to light touch (p < 0.05). Finger soaking enhanced the effects of light fingertip touch on reducing body sway only in those children with developmental coordination disorder (p < 0.05). CONCLUSION: Finger soaking can be used as a rehabilitation strategy for promoting sensitivity to light touch, as well as for enhancing the effects of light fingertip touch in reducing body sway in children with developmental coordination disorder.
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Dedos/fisiologia , Transtornos das Habilidades Motoras/reabilitação , Equilíbrio Postural/fisiologia , Tato/fisiologia , Criança , Feminino , Humanos , MasculinoRESUMO
The present study was designed to investigate the effects of lipoxin A4 (LXA4) on traumatic brain injury (TBI) and to analyze the possible mechanism. Outcome parameters consist of blood-brain barrier (BBB) breakdown, brain edema and lesion volume. Using western blot and quantitative real-time PCR, we examined the levels of pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6) and activation of mitogen-activated protein kinases (MAPKs) (including ERK, JNK, p38) following TBI. To investigate the cell types in which the LXA4 receptor (ALXR) staining was localized, brain sections pulsed with ALXR were subjected to immunofluorescence staining with antibodies against cell type-specific antigens. Our findings show that LXA4 decreases BBB permeability, attenuates brain edema, and reduces TBI-induced lesion volume. In addition, LXA4 inhibits TBI-induced elevation of mRNA and protein levels of pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6). In the injured cortex at 24h post-TBI, the phosphorylated-ERK (p-ERK) and -JNK (p-JNK), but not -p38 (p-p38) levels were increased. The p-ERK and p-JNK production were attenuated by their respective inhibitors (PD98059 and SP600125), as well as LXA4. Moreover, ALXR was found to label more GFAP positive cells, whereas few CD11b-positive cells were labeled by ALXR within the layers of the injured cortex at 24h post-TBI. The activation of ALXR in astrocytes was partially enhanced by LXA4 treatment. Taken together, these data indicate that TBI activates pro-inflammatory cytokines, the MAPK pathways together with ALXR in astrocytes, and these mechanisms may be exploited by pharmacological interventions.
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Lesões Encefálicas/tratamento farmacológico , Citocinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Lipoxinas/uso terapêutico , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Análise de Variância , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/fisiopatologia , Edema Encefálico/prevenção & controle , Lesões Encefálicas/patologia , Antígeno CD11b/metabolismo , Citocinas/genética , Modelos Animais de Doenças , Lipoxinas/farmacologia , Masculino , Camundongos , Fosforilação/efeitos dos fármacos , RNA Mensageiro/metabolismoRESUMO
Acute membrane damage due to traumatic brain injury (TBI) is a critical precipitating event. However, the subsequent effects of the mechanical trauma, including mitochondrial and lysosomal membrane permeability (MOMP and LMP) remain elusive. The main objective of the current study was to assess the role of a putative membrane-resealing agent poloxamer 188 (P188) in MOMP and LMP in response to a well-defined mechanical insult. Using an in vitro cell shearing device (VCSD), mechanical injury resulted in immediate disruption of membrane integrity in cultured primary neurons, and neurons were treated with P188 or a cathepsin B inhibitor (CBI) after VCSD 10 min. The protective effect of P188 on cultured primary neurons was first detected visually with a light microscope, and measured by MTT assay and LDH assay. The validity of monitoring changes in mitochondrial membrane potential (ΔΨm) was measured by JC-1 staining, and Western blot for cytochrome c and truncated Bid (tBid) in purified mitochondria was also performed. In addition, lysosomal integrity was detected by blotting for cathepsin B and tBid in purified lysosomes. Our results showed post-injury P188 treatment moderated the dissipation of ΔΨm in mitochondria, and inhibited VCSD-induced cytochrome c release from mitochondria as well as cathepsin B from lysosomes. Cathepsin B inhibition (CBI) could also increase cell viability, maintain mitochondrial membrane potential, and repress VCSD-induced release of cytochrome c from mitochondria to cytosol. Both P188 and CBI treatment decreased the cytosolic accumulation of tBid in supernatant of purified lysosomes, and the amount of mitochondrial localized tBid. These data indicate injured neurons have undergone mitochondrial and lysosomal membrane permeability damage, and the mechanism can be exploited with pharmacological interventions. P188's neuroprotection appears to involve a relationship between cathepsin B and tBid-mediated mitochondrial initiation of cell death.
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Lesões Encefálicas/patologia , Lisossomos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Poloxâmero/farmacologia , Animais , Western Blotting , Lesões Encefálicas/metabolismo , Células Cultivadas , Membranas Intracelulares/efeitos dos fármacos , Membranas Intracelulares/metabolismo , Membranas Intracelulares/patologia , Lisossomos/metabolismo , Lisossomos/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Neurônios/metabolismo , Neurônios/patologia , Permeabilidade , Ratos , Ratos Sprague-DawleyRESUMO
<p><b>OBJECTIVE</b>To study the in vitro expression of 6 novel missense mutations (R270G, P275A, F121L, A156P, E183G, I324N) and a previously described R408Q mutation of phenylalanine hydroxylase (PAH) gene and explore the genotype-phenotype correlation through comparison of protein levels and residual enzyme activities.</p><p><b>METHODS</b>Seven expression vectors containing PAH cDNA were constructed with a site-directed mutagenesis kit. The plasmids were extracted and sequenced to confirm the target mutations. pcDNA3.0 containing PAH cDNA was transfected into COS-7 cells and total proteins were extracted 48 h after transfection. The quantities of proteins and residual enzyme activities of the 7 mutants were assessed with the wild-type PAH gene as reference.</p><p><b>RESULTS</b>Relative quantities of PAH proteins for R270G, P275A, F121L, A156P, E183G, I324N and R408Q were 10.5%, 56.6%, 54.3%, 8.7%, 8.5%, 67.3% and 85.4%, respectively. The residual enzyme activities were 7.7%, 27.6%, 19.0%, 10.4%, 9.1%, 50.6% and 40.2%, respectively.</p><p><b>CONCLUSION</b>PAH residual enzyme activities of 7 PAH mutants were all significantly reduced.</p>
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Animais , Humanos , Sequência de Aminoácidos , Células COS , Chlorocebus aethiops , Estudos de Associação Genética , Métodos , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Fenilalanina Hidroxilase , Genética , Alinhamento de SequênciaRESUMO
Jumonji Domain Containing 2A (JMJD2A) may be a cancer-associated gene involved in human breast cancer. With a view to investigating expression of JMJD2A in human breast cancer and benign lesion tissues as well as relationship between JMJD2A and tumor related proteins, histological and immunohistochemical analysis, Western blot and quantitative real-time PCR in infiltrating duct carcinoma and fibroadenoma for JMJD2A and immunohistochemical analysis and quantitative real-time PCR in infiltrating duct carcinoma for tumor related proteins (ARHI, p53, ER, PR and CerbB-2) were performed. Histological examination validated the clinical diagnosis. The JMJD2A positive rate of infiltrating duct carcinoma was significantly higher than fibroadenoma by immunohistochemical analysis. The mean optical density of JMJD2A in infiltrating duct carcinoma was higher than fibroadenoma by western blot. JMJD2A mRNA level in infiltrating duct carcinoma was higher than fibroadenoma by quantitative real-time PCR. Spearman correlation analysis revealed that the expression of JMJD2A was associated with ARHI, p53 and ER from immunohistochemical results respectively. Pearson correlation analysis revealed that the expression of JMJD2A was associated with ARHI, p53 and ER from quantitative real-time PCR results respectively. Expression of JMJD2A in infiltrating duct carcinoma was higher, and associated with ARHI, p53 and ER. The results may take JMJD2A as a potential diagnostic and therapeutic target in human breast cancer.
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Neoplasias da Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Linhagem Celular Tumoral , Feminino , Fibroadenoma/metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Histona Desmetilases com o Domínio Jumonji/biossíntese , Receptor ErbB-2/biossíntese , Receptores de Estrogênio/biossíntese , Receptores de Progesterona/biossíntese , Proteína Supressora de Tumor p53/biossíntese , Proteínas rho de Ligação ao GTP/biossínteseRESUMO
<p><b>OBJECTIVE</b>To investigate the expression and significance of Clusterin in normal prostate, benign prostate hyperplasia (BPH) and prostate cancer.</p><p><b>METHODS</b>Clusterin expression in samples of 12 normal prostate, 15 BPH, and 56 prostate cancer were studied by immunohistochemical stain.</p><p><b>RESULTS</b>Of 83 cases, 67 are positive or weak positive (81%). The rate of positive or weak positive for normal prostate, BPH and prostate cancer was 17% (2/12), 73% (11/15), and 96% (54/56) respectively. The expression level of Clusterin in prostate cancer was much higher than in normal prostate (t = 8.82, P < 0.01). BPH (t = 7.63, P < 0.01) was related positively with pathological grade (r = 0.649, P < 0.01) and stage (r = 0.609, P < 0.01) of prostate cancer.</p><p><b>CONCLUSION</b>Clusterin may play an important role in the biological characteristics of prostate cancer by the anti-apoptosis pathway.</p>
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Feminino , Humanos , Masculino , Apoptose , Clusterina , Metabolismo , Fisiologia , Imuno-Histoquímica , Próstata , Metabolismo , Hiperplasia Prostática , Metabolismo , Neoplasias da Próstata , Metabolismo , PatologiaRESUMO
<p><b>OBJECTIVE</b>To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).</p><p><b>METHODS</b>Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.</p><p><b>RESULTS</b>The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.</p><p><b>CONCLUSION</b>The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.</p>