Aerobic Glycolysis Induced by mTOR/HIF-1α Promotes Early Brain Injury After Subarachnoid Hemorrhage via Activating M1 Microglia.

M1 microglial activation is crucial for the pathogenesis of early brain injury (EBI) following subarachnoid hemorrhage (SAH), and there is growing evidence that glucose metabolism is frequently involved in microglial activation. However, the molecular mechanism of glycolysis and its role in M1 microglial activation in the context of EBI are not yet fully understood. In this study, firstly, the relationship between aerobic glycolysis and M1 microglial activation as well as SAH-induced EBI was researched in vivo. Then, intervention on mammalian target of rapamycin (mTOR) was performed to investigate the effects on glycolysis-dependent M1 microglial activation and EBI and its relationship with hypoxia-inducible factor-1α (HIF-1α) in vivo. Next, Hif-1α was inhibited to analyze its role in aerobic glycolysis, M1 microglial activation, and EBI in vivo. Lastly, both in vivo and in vitro, mTOR inhibition and Hif-1α enhancement were administered simultaneously, and the combined effects were further confirmed again. The results showed that aerobic glycolysis and M1 microglial polarization were increased after SAH, and glycolytic inhibition could attenuate M1 microglial activation and EBI. Inhibition of mTOR reduced glycolysis-dependent M1 microglial polarization and EBI severity by down-regulating HIF-1α expression, while enhancement had the opposite effects. Blockading HIF-1α had the similar effects as suppressing mTOR, while HIF-1α agonist worked against mTOR antagonist when administered simultaneously. In conclusion, the present study showed new evidence that aerobic glycolysis induced by mTOR/HIF-1α might promote EBI after SAH by activating M1 microglia. This finding provided new insights for the treatment of EBI.

Osteoprotegerin inhibit osteoclast differentiation and bone resorption by enhancing autophagy via AMPK/mTOR/p70S6K signaling pathway in vitro.

Osteoclasts are highly differentiated terminal cells formed by fusion of hematopoietic stem cells. Previously, osteoprotegerin (OPG) inhibit osteoclast differentiation and bone resorption by blocking receptor activator of nuclear factor-κB ligand (RANKL) binding to RANK indirect mechanism. Furthermore, autophagy plays an important role during osteoclast differentiation and function. However, whether autophagy is involved in OPG-inhibited osteoclast formation and bone resorption is not known. To elucidate the role of autophagy in OPG-inhibited osteoclast differentiation and bone resorption, we used primary osteoclast derived from mice bone marrow monocytes/macrophages (BMM) by induced M-CSF and RANKL. The results showed that autophagy-related proteins expression were upregulated; tartrate-resistant acid phosphatase-positive osteoclast number and bone resorption activity were decreased; LC3 puncta and autophagosomes number were increased and activated AMPK/mTOR/p70S6K signaling pathway. In addition, chloroquine (as the autophagy/lysosome inhibitor, CQ) or rapamycin (as the autophagy/lysosome inhibitor, Rap) attenuated osteoclast differentiation and bone resorption activity by OPG treatment via AMPK/mTOR/p70S6K signaling pathway. Our data demonstrated that autophagy plays a critical role in OPG inhibiting osteoclast differentiation and bone resorption via AMPK/mTOR/p70S6K signaling pathway in vitro.

CT diagnosis and differential diagnosis of renal cell carcinoma associated with Xp11.2 translocation/TFE gene fusions / 医学研究生学报

Objective XPll.2/TFE RCC is an independent subtype of renal cell carcinoma, which is rare. The aim of this study is to investigate the CT features of Xp11.2/TFE RCC and improve the accuracy of diagnosis. MethodsCT findings of 30 cases from August 2009 to September 2017 of Xp11.2/TFE RCC from Eastern War Zone General Hospital, including location, density, edge, enhancement degree, lymphatic metastasis and others. They were analyzed respectively and compared with those of ccRCC. ResultsThe differences in CT values between tumors and renal cortex and renal medulla was statistically significant different phases (PP< 0.01). Most of patients with Xp11.2/TFE RCC are younger, about (36.4±17.7) years old. Females are more common, accounting for about 70% compared with ccRCC(50%). CT plan scan showed slightly higher density, about (45.2±8.9)HU vs (34.1±4.4)HU, high calcification rate, about 46.7% % vs 10.0% and CT scan with contrast agent showed gradual enhancement. The difference all were statistically significant(P<0.05). Conclusion XPll.2/ has certain CT characteristics. Combined with CT and clinical manifestation of patients, it is helpful to improve the accuracy of preoperative diagnosis.

Effects of Ringer's solution with different concentrations of alcohol on biphasic compound action potentials of frog sciatic nerve trunk / 中国应用生理学杂志

OBJECTIVE@#To quantitatively investigate the effects of Ringer's solution with different concentrations of alcohol (1%～80%) on biphasic compound action potentials (AP) from frog sciatic nerve trunk, and their recoveries from alcohol effects.@*METHODS@#Individual segments of frog sciatic nerve trunk with a length of 6 to 8 cm were prepared. Ringer's solution with different concentrations of alcohol (0%, 1%, 2%, 4%, 8%, 16%, 32%, 48%, 64% and 80%) was applied onto the segment of the trunk between the stimulus and ground electrodes via an agent reservoir which was newly armed in a nerve trunk shielded chamber for 5 minutes. The nerve trunk was respectively electro-stimulated to generate the biphasic compound AP which was recorded using the experimental system of BL-420F. This was followed by 5 times washout plus 5 min administration with Ringer's solution before recovery recording of AP.@*RESULTS@#Compared to normal Ringer's solution, Ringer's solution with alcohol at ≤4% did not have dramatic impacts on the AP amplitude and conduction velocity, while Ringer's solution with alcohol at ≥8% there was significant decrease in these two parameters. Ringer's solution with alcohol at the conentrations of 16%, 32% and ≥48% could prevent a small proportion (30%), a large proportion (90%) and all (100%) of sciatic nerve trunks, respectively, from generating AP. Washout with normal Ringer's solution after alcohol application at the concentration of ≤32%, AP could totally recover to normal status. While alcohol at the concentration of 48%, 64% and 80%, the probabilities to regenerate APs were 90%, 40% and 0%, and the AP amplitudes were decreased to 60%, 36% and 0%, respectively. After washout, AP conduction velocity showed no difference with alcohol at the concentration of ≤8% when compared with that before washout, while it could not be recovered to normal under alcohol at ≥16%.@*CONCLUSION@#Ringer's solution with different concentrations of alcohol exerts different effects on biphasic compound AP amplitude and conduction velocity. Hopefully, our findings could be helpful for the alcoholic usage and its recovery from alcoholic damage.

ROSIER scale is useful in an emergency medical service transfer protocol for acute stroke patients in primary care center: A southern China study

Objective: The aim of the study is to validate whether the Recognition Of Stroke In the Emergency Room (ROSIER) scale can be used by general practitioners (GPs) in an emergency medical service (EMS) protocol to transfer stroke patients from primary care center to advanced hospital with acute stroke center. Methods: GPs prospectively performed the ROSIER scale and the Cincinnati Prehospital Stroke Scale (CPSS) on suspected stroke patients as a transfer protocol. All patients were immediately transferred to the Level-II hospital for further treatment. Results: 468 of the 512 suspected stroke patients met the inclusion criteria in this study. The ROSIER scale showed a diagnostic sensitivity of 83.13% (95% confidence intervals [CI] 79.74-86.52%) and specificity of 80.88% (95% CI 77.32- 84.44%). The CPSS showed a diagnostic sensitivity of 78.01% (95% CI 74.26-81.76%) and specificity of 70.59% (95% CI 66.46-74.72%). The Kappa statistic value of the ROSIER scale and the CPSS were 0.601 and 0.454, respectively. The area under the curve (AUC) of ROSIER scale was large than the CPSS (AUC 0.855 vs. 0.791). However, the difference was not significantly different. Conclusions: This study suggest that ROSIER and CPSS could be used in an EMS protocol to transfer stroke patients from a primary care center to an advanced hospital offering thrombolysis service

The effects and mechanism of estrogen on rats with Parkinson's disease in different age groups.

OBJECTIVE: In order to investigate the effect and mechanism of estrogen in rotenone-induced Parkinson's disease (PD) rats in different age groups. METHODS: we established rat models of PD by rotenone at different interventions. Then, behavioral tests, immunohistochemistry, western blot, high-performance liquid chromatography-electrochemical detector (HPLC-ECD) and electron microscopy were performed. RESULTS: Results revealed the following: (1) Rotenone significantly reduced rotarod latencies in senile rats, prolonged their climbing pole time, and decreased TH positive cells, DA and its metabolite, DOPAC. Estrogen ameliorated this effect, in which weaker effects were observed in younger rats compared with older rats. (2) Rotenone increased the expression of LC3-II in older rats, but estrogen and tamoxifen did not show the same effect. (3) Rotenone increased the number of autophagosomes, but estrogen increased the proportion of autolysosomes/autophagosomes in the rotenone-treated group. (4) U0126 could reduce the number of autophagosomes in the rotenone-treated group, but this did not change the proportion of autolysosome/autophagosome in combining rotenone with the estrogen group. Rapamycin did not increase the number of autophagosomes in the rotenone-treated group, but combining rapamycin with estrogen and rotenone was able to further increase the proportion of autolysome/autophagosomes. Therefore, we speculate that the senile rat model of PD was more reliable than that in young rats. CONCLUSIONS: In addition, estrogen could promote autophagy maturation through the ERK pathway, and had an obvious therapeutic effect on the rat model of PD.

Upregulation of cell surface estrogen receptor alpha is associated with the mitogen-activated protein kinase/extracellular signal-regulated kinase activity and promotes autophagy maturation.

Recently, accumulating evidence has implicated the dysregulation of autophagy as underlying the pathophysiology of several neurodegenerative diseases. The human neuronal cell line SH-SY5Y was exposed to 1-Methyl-4-phenylpyridinium (MPP(+)). The mechanism is that the sustained activation of the MAPK/ERK pathway by MPP(+) alters autophagy selectively at the maturation step, significant increasing in autophagy formation and delaying in autophagy degradation in SHSY5Y cells. In this study, we provided evidences that estrogen was capable of promoting SHSY5Y cells survival in MPP(+)-treated group. In particular, the up-regulation of mERα, but not mERß, was associated with a rapid and transient activation of ERK phosphorylation compatible with promoting autophagy maturation. The up-regulation of mERα changed the sustained activation of ERK phosphorylation in MPP(+)-treated group into a temporary activation. Taken together, these findings strongly support that the expression of mERα promotes the maturation of autophagosomes into functional autolysosomes by regulating ERK, determining SHSY5Y cells survival.

MRI Appearances and Clinical Analysis of Lateral Ventricular Subependymomas / 天津医药

Objective Analysis of MRI image of lateral ventricular subependymomas to improve our recognition of radiology manifestation of the disease. Methods Clinical data and MRI findings of 6 patients with subependymomas proved by surgical pathology, observed from January 2007 to June 2013, were analyzed retrospectively. Results Six patients (4 males and 2 females) presented with headache and dizziness. Among these 6 patients, tumor developed in right lateral ventri-cle in three patients, in left lateral ventricle in 2 patients and in frontal of dual lateral ventricle in one patient. In MRI image, tumors showed iso-intense to hypointense to normal white matter on T1-weighted and DWI images, hyperintense on T2-weighted and FLAIR. Mild or no enhancement was noted in most cases. MRS showed findings consistent with low-grade tu-mor, with a normal choline peak and depressed N-acetyl-aspartate peak. Conclusion Lateral ventricular subependymo-mas have characteristic MRI features and multi-direction images are helpful for diagnosis.

Promotion of autophagy at the maturation step by IL-6 is associated with the sustained mitogen-activated protein kinase/extracellular signal-regulated kinase activity.

Increased autophagic vacuoles (AVs) occur in injured or degenerating neurons, under both developmental and pathological situations. Although an induced autophagy has been shown in inflammation response to cell factors, the underlying mechanism(s) remain(s) unknown. Here, we show that both cell factor IL-6 and environmental toxin MPP(+) promote the formation of vacuolation in SHSY5Y cells. By electron and immunofluorescent microscopy analyses, we showed that these structures are acid autolysosomes, containing cellular debris, and labeled by LC3 or LAMP1, markers of autophagosomes or lysosomes, respectively. Combining MPP(+) and IL-6 do not further increase vacuolation of SHSY5Y cells, and the vacuolation is less than that in the MPP(+)-treated group. MPP(+)-induced vacuolation results from significant increase in autophagy formation and delay in autophagy degradation, in relation to a decline of the lysosomal activity of arylsulfatase A. At molecular level, we show that this defect in autolysosomal maturation is independent of mammalian target of rapamycin and p38 inhibitions. Most importantly, we provide the first evidence that activation of ERK pathway is sufficient to commit cell to autophagic vacuolation. The sustained activation is required for MPP(+) to disrupt the autophagic pathway. IL-6 also induces a temporary and significant activation of ERK, but not sustained activation, and change sustained activation in MPP(+)-treated group into temporary activation. Taken together, these findings strongly support that IL-6 promotes the maturation of autophagosomes into functional autolysosomes by regulating ERK.

The multiplicity analysis of massive hemothorax after thoracic operation / 中国医师进修杂志

Objective To explore multi-causes and therapy of massive hemothorax after thoracic operation.Methods Sixty-six patients suffered from massive hemothorax after thoracic operation.All of them were executed conservative treatments for postoperative hemothorax.The noneffeetive cases were executed re-exploration.The relationship of area of residual cavity,fluctuation of intrapleural pressure and volume of hemothorax were analyzed between lobectomy in 30 eases and wedge,segmental or no excision of lung in 24 cases in 24 h postoperation.Results Thirty-two of 66 cases being executed conservative treatments were suteessful,2 cases were dead,while 32 cases were executed re-exploration,and 29 of them were cured.but 1 case of them dead,and 2 cases suffered from bronchial fistula,who were cured by thoracoplasty.The operations of wedge,segmental or no excision of lung in 24 cases were compared with lobectomy in 30 cases.It Was proved that the former had the smaller area of residual cavity,the lower intrapleural pressure.and the less volume of hemothorax(P＜0.05).Conclusions The multiplicity analysis of massive hemothorax after thoracic operation are flucmafion of intrapleural pressure after operation,intracavitary suction with negative pressure,rise of pressure in microcirculation at wound,abnormality of blood coagulation function and so on.It can reduce complications that proper therapy is timely performed,and even avoid of re-exploration.

Efficacy of rabdosia rubescens in the treatment of gingivitis / 华中科技大学学报（医学）（英德文版）

This study evaluated the efficacy of rabdosia rubescens against gingivitis and compared the therapeutic efficacy of different dosage forms of rabdosia rubescens. A multi-center, randomized, double-blind, double-simulation, positive-controlled and parallel trial was conducted. A total of 136 patients exhibiting clinical symptoms of gingivitis were enrolled. The subjects were randomly assigned to two groups: test group (n=67), in which rabdosia rubescens drop pill (960 mg) and 4 tablets of simulation agent of rabdosia rubescen were orally given to the subjects three times a day for 5 days; and control group (n=69), in which the subjects were administered the tablets of rabdosia rubescens (1000 mg) and 24 drop pills of simulation agent of rabdosia rubescens thrice daily for 5 days. The experimental protocols and diagnostic criteria were established by expert panel prior to the experiment. The clinical symptoms were graded according to the severity of the disease and quantified. The total scores and scores for each clinical symptom of gingivitis were assessed at baseline and on the 6th day post-treatment. The therapeutic efficacy was compared between the two groups and in each group itself before and after the treatment. The results showed that in the two groups, the subjects who were given rabdosia rubescens, drop pill or tablet, had a decrease in total scores and scores for each clinical symptom when compared with those before treatment (P<0.01). There was significant difference in the therapeutic efficacy between the test group and the control group with the efficacy rate being 92.54% and 79.71% respectively (P<0.05). It was concluded that rabdosia rubescens showed great promise in treating gingivitis. And rabdosia rubescens drop pill was more efficacious than rabdosia rubescens tablet.

The effect of water extracts of Euphorbia hirta on cartilage degeneration in arthritic rats

The effect of water extracts of Euphorbia hirta on the histological features and expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the rat articular cartilage was investigated. Arthritis was induced in rats using Freund's Complete Adjuvant containing heat-killed M. tuberculosis, and treated with water extracts of E. hirta. Paraffin tissue sections of the arthritic joints were evaluated. The extent of cartilage degeneration was found to be greatest in rats treated with the highest dosage of E. hirta, followed by rats in the untreated group. Rats treated with the intermediary and low dosages of Euphorbia hirta showed improved histology. MMP-13 levels were found to be decreased with decreasing dosages of E. hirta. TIMP-1 levels were found to increase with decreasing dosages of E. hirta. MMP-3 levels fluctuated without any appreciable pattern. Low dosages of E. hirta seem to be beneficial in reducing cartilage degeneration in cases of arthritis.

Selenoprotein P mRNA expression in human hepatic tissues.

AIM: To investigate the expression of selenoprotein P mRNA (SePmRNA) in tissues of normal liver, liver cirrhosis and hepatocellular carcinoma (HCC), and its relationship with HCC occurrence and development. METHODS: The expression of SePmRNA in tissues of normal liver, liver cirrhosis and HCC were detected by in situ hybridization using a cDNA probe. RESULTS: The enzyme digesting products of PBluescript-Human Selenoprotein P were evaluated by electrophoresis. The positive expression of SePmRNA was found in the tissues of normal liver, liver cirrhosis and HCC. The expression of SeP mRNA was found in hepatic interstitial substance, especially in endothelial cells and lymphocytes of vasculature. The positive rate of SePmRNA in normal liver tissue was 84.6% (11/13) and the positive signals appeared in the nucleus and cytoplasm, mostly in the nucleolus, and the staining granules were larger in the nucleolus and around the nucleus. The positive rate of SePmRNA in liver cirrhosis tissue was 45.0% (9/20) and the positive signals were mainly in the nucleolus and cytoplasm, being less around the nucleus and inner nucleus than that in normal liver tissue. The positive rate of SePmRNA in HCC tissue was 30.0% (9/30) and the positive signals were in the cytoplasm, but less in the nucleus. CONCLUSION: SePmRNA expression in the tissues of normal liver and HCC is significantly different (84.6% vs 30.0%, P=0.003), suggesting that SeP might play a role in the occurrence and development of HCC.

Diet-induced obesity affects testis development in pubertal rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of diet-induced obesity on the developmental process of testes in pubertal rats.</p><p><b>METHODS</b>Eighty 21-day-old male SD rats were randomly divided into a control group (n=32) and an experiment group (n=48), and fed respectively on a normal diet and a high-fat diet. And changes in the body weight, Lee's index, testis weight and epididymis weight were measured at the end of the 3rd, 4th, 5th and 6th week after the treatment, that is, when the rats were 6, 7, 8 and 9 weeks old. The concentrations of testosterone and estradiol were determined by Access immunoassay system and the morphological alterations in testis development observed by HE staining.</p><p><b>RESULTS</b>The body weight of the high-fat group obviously increased at the end of the 3rd week (P < 0.05), 26.6% heavier than that of the control by the end of the 6th week (P < 0.01), and Lee's index was also obviously increased (P < 0.01). Compared with the controls, the testicular coefficient declined in the high-fat group at the end of the 5th and 6th week (P < 0.05), plasma TG and TC remarkably increased, the testosterone level obviously decreased (P < 0.05), estradiol concentration lowered at the end of the 3rd, 4th and 5th week but dramatically increased at the 9th, with significant difference between the two groups (P < 0.01). Microscope examination showed that spermatogenic epithelial cells were arranged in disorder, the spermatogenic cell layers reduced and the number of mature sperms reduced.</p><p><b>CONCLUSION</b>High-fat diet can induce nutritional obesity in pubertal rats, which in turn may lead to the underdevelopment of the testis and the abnormal level of gonadal hormones.</p>

Expression and clinical significance of TAp73alpha, p53, PCNA and apoptosis in hepatocellular carcinoma.

AIM: To study the prognostic role of TAp73alpha, p53, proliferating cell nuclear antigen (PCNA) and apoptosis in patients with hepatocellular carcinoma (HCC) after surgical tumor ablation. METHODS: Forty-seven human resected HCC tissues and 42 adjacent non-cancerous tissues were studied with 10 normal liver tissues as control group. TAp73alpha, p53, and PCNA were detected with Elivision immunohistochemistry. Terminal deoxynucleotidyl transferase (TdT)-mediated d-UTP-biotin nick-end labeling (TUNEL) method was used to detect the apoptosis cells. All clinical and pathological materials were analyzed by SPSS10.0 statistical package. RESULTS: TAp73alpha overexpressed in HCC tissues (36.2%) when compared with adjacent non-cancerous tissues (2.38%, P<0.005) and normal liver tissues (0, P<0.01). Mutant type p53 (mt-p53) overexpressed in HCC tissues (38.3%) when contracted with adjacent non-cancerous tissues (16.7%, P<0.05) and normal liver tissues (0, P<0.01). Proliferation index (PI) level in HCC tissues was significantly higher than that in adjacent non-cancerous tissues (30.34%+/-4.46% vs 27.88%+/-5.89%, t, P = 0.028). Apoptosis index (AI) level in HCC tissues was higher than that in adjacent non-cancerous tissues (8.62%+/-2.28% vs 7.38%+/-2.61%, t, P = 0.019). Expression of TAp73alpha was associated with lymph node metastasis and mt-p53, with r = 0.407 and 0.265, respectively. Expression of mt-p53 was associated with Edmondson's stage and AFP, with r = 0.295 and -0.357, respectively. In Kaplan-Meier univariant analysis, TAp73alpha, AFP, TNM stage, portal vein invasion, liver membrane invasion and HBsAg correlated with prognosis (log rank, P = 0.039, 0.012, 0.002, 0.000, 0.014, 0.007, respectively). Multivariant Cox regression analysis showed that TAp73alpha, AFP, TNM stage, portal vein invasion, liver membrane invasion and age were independent factors of prognosis. CONCLUSION: These results suggest that TAp73alpha can be used as a prognostic indicator of patients with HCC undergoing surgical tumor ablation. AFP, TNM, portal vein invasion, liver membrane invasion and age also have a potency of predicting the prognosis of HCC.

Effects of depression on parameters of cell-mediated immunity in patients with digestive tract cancers.

AIM: To evaluate the effects of depression on parameters of cell-mediated immunity in patients with cancers of the digestive tract. METHODS: One hundred and eight adult patients of both sexes with cancers of the digestive tract admitted between March 2001 and February 2002 in the Department of Medical Oncology, First Affiliated Hospital of Xi'an Jiaotong University were randomly enrolled in the study. The Zung self-rating depression scale (SDS), Zung self-rating anxiety scale (SAS), numeric rating scale (NRS) and social support rating scale (SSRS) were employed to evaluate the degree of depression and their contributing factors. In terms of their SDS index scores, the patients were categorized into depression group (SDS> or =50) and non-depression group (SDS<50). Immunological parameters such as T-lymphocyte subsets and natural killer (NK) cell activities in peripheral blood were determined and compared between the two groups of patients. RESULTS: The SDS index was from 33.8 to 66.2 in the 108 cases, 50% of these patients had a SDS index more than 50. Similarly, the SAS index of all the patients ranged from 35.0 to 62.0 and 46.3% of the cases had a SAS index above 50. Cubic curve estimation showed that the depression was positively correlated with anxiety and negatively with social support. Furthermore, the depression correlated with the tumor type, which manifested in a descending order as stomach, gallbladder, pancreas, intestine, esophagus, duodenum and rectum, according to their correlativity. Step-wise regression analysis suggested that hyposexuality, dispiritment, agitation, palpitation, low CD56 and anxiety were the significant factors contributing to depression. More severe anxiety (49.7 +/- 7.5 vs 45.3 +/- 6.9, P<0.05), pain (6.5 +/- 2.8 vs 4.6 +/- 3.2, P<0.05), poor social support (6.8 +/- 2.0 vs 7.6 +/- 2.1, P<0.05), as well as decline of lymphocyte count (0.33 +/- 0.09 vs 0.39 +/- 0.87, P<0.05) and CD56 (0.26 +/- 0.11 vs 0.29 +/- 0.11, P<0.05) were noted in the depression group compared with those of the non-depression patients. However, fewer obvious changes in CD4/CD8 ratio and other immunological parameters were found between the two groups. CONCLUSION: Depression occurs with a high incidence in patients with cancers of the digestive tract, which probably is not the sole factor leading to the impairment of immunological functions in these cases. However, comprehensive measures including psychological support should be taken in order to improve the immunological function, quality of life and clinical prognosis of these patients.

Cloning of human 15ku selenoprotein gene from H9 T cells.

AIM: To clone human 15ku selenoprotein gene. METHODS: H9 human T cells were cultured in RPMI1640 medium supplemented with 100 mL /L fetal calf serum. mRNA was isolated from the cells. cDNA library was constructed by RT-PCR. The human 15ku selenoprotein gene was obtained by PCR and cloned into T vector and sequenced. RESULTS: A unique cDNA fragment about 1 244 bp was obtained. Sequence analysis identified an open reading frame within the cDNA. The gene had an in-frame TGA, which encoded selenocysteine (Sec), and a 3'-UTR SECIS element, which was required for synthesis of selenoprotein. The predicted protein molecular mass was about 15ku (162 residues). The result was identical with human liver 15ku selenoprotein gene published in Genbank. CONCLUSION: Human 15ku selenoprotein gene can be successfully obtained from T cell line.