Discovery of exogenous ligands for orphan receptor BRS-3 from Chinese herbs / 中国中药杂志

Bombesin receptor subtype-3(BRS-3) is an orphan receptor in the bombesin receptor family. Its signal transduction mechanism and biological function have attracted much attention. Seeking the ligand for BRS-3 is of great significance for exploring its function. Considering the fact that the activation of BRS-3 receptor can induce the change in intracellular Ca~(2+) concentration, the fluo-rometric imaging plate reader(FLIPR) was utilized for ligand screening at the cellular level. Among more than 400 monomeric compounds isolated from Chinese herbs, yuanhunine from Corydalis Rhizoma and sophoraisoflavanone A and licoriphenone from Glycyrrhizae Radix et Rhizoma antagonized BRS-3 to varying degrees. It was confirmed in HEK293 cells expressing BRS-3 that yuanhunine, sophoraisoflavanone A, and licoriphenone inhibited the calcium current response after the activation of BRS-3 by [D-Phe~6,β-Ala~(11),Phe~(13),Nle~(14)]bombesin-(6-14) in a dose-dependent manner with the IC_(50) values being 8.58, 4.10, and 2.04 μmol·L~(-1), respectively. Further study indicated that yuanhunine and sophoraisoflavanone A exhibited good selectivity for BRS-3. In this study, it was found for the first time that monomers derived from Chinese herbs had antagonistic activity against orphan receptor BRS-3, which has provided a tool for further study of BRS-3 and also the potential lead compounds for new drug discovery. At the same time, it provides reference for the research and development of innovative drugs based on the active ingredients of Chinese herbs.

Application of Methylprednisolone Sodium Succinate Combined with Tropisetron in Prevention of Nausea and Vomiting under Microvascular Decompression of Hemifacial Spasm / 中国医学科学院学报

Objective To evaluate the effect of methylprednisolone sodium succinate combined with tropisetron on postoperative nausea and vomiting(PONV)under microvascular decompression of hemifacial spasm.Methods From January to June 2019,485 patients undergoing microvascular decompression for facial spasm at Department of Neurosurgery,Peking University People's Hospital were randomly assigned into two groups with random number table method.For group A(n=242),2 ml saline was administrated by intravenous drip before induction and 5 mg tropisetron after operation.For group B(n=243),40 mg methylprednisolone sodium succinate was administrated by intravenous drip before induction and 5 mg tropisetron after operation.The anesthesia time,operation time,and incidence of PONV in 0-24 h and 24-48 h were recorded for the comparison of the remedial treatment rate of nausea and vomiting between the two groups.Results There was no significant difference in age,gender,smoking history,body mass index value,American Society of Anesthesiologists score,medical history,surgical side,PONV history,operation time or anesthesia time between the two groups(all P > 0.05).The incidence of PONV in group A was 35.5% and 18.2% during 0-24 h and 24-48 h,respectively,which was significantly higher than that(18.5%,χ

Clinical value of D-dimer and CRP in evaluating the prognosis of 2019 novel coronavirus-infected pneumonia / 医学研究生学报

ObjectiveTo investigate the relationship between the levels of D-dimer and inflammatory factors C-reactive protein（CRP）and prognosis in patients with 2019 novel coronavirus-infected pneumonia（COVID-19）.Methods The clinical data of a total of 242 patients with COVID-19 who were treated in hospital from February 4th 2020 to February 18th 2020 were analyzed retrospectively. According to the classification standard,the patients with COVID-19 were divided into common patients(131 cases), severe patients（88 cases）, and critical patients（23 cases）. The difference between the levels of D-dimer and CRP in patients with pneumonia of different severity and clinical outcomes was compared and the correlation between D-dimer and CRP was analyzed.ResultsThe levels of D-dimer and CRP in severe and critical patients were significantly higher than those in common patients(P<0.05). The levels of CRP in critical patients were significantly higher than those in severe patients(P<0.05). These two indicator levels of patients who died of COVID-19 within 30 dayswere significantly higher than those who survived. Pearson correlation analysis showed that the levels of D-dimer were positively correlated with the levels of CRP(r=0.649，P<0.05).ConclusionD-dimer and CRP are highly expressed in severe and critical patients, and the severe abnormality of the two indicators in the early stage of COVID-19 predicted the poor prognosis. D-dimer and CRP have certain clinical value in evaluating the severity and prognosis of COVID-19.

MiR-182-5p Knockdown Targeting PTEN Inhibits Cell Proliferation and Invasion of Breast Cancer Cells

PURPOSE: Breast cancer (BC) is one of the most common malignant tumors, affecting a significant number of women worldwide. MicroRNAs (miRNAs) have been reported to play important roles in tumorigenesis. The aim of this study was to determine the roles of miR-182-5p in BC progression. MATERIALS AND METHODS: The expressions of miR-182-5p and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) were measured in BC tissues and cells by quantitative real-time polymerase chain reaction or Western blot. Cell proliferation and invasion were detected by cell counting kit-8 assay and trans-well assay, respectively. The interaction between miR-182-5p and PTEN was probed by bioinformatics analysis, luciferase activity, and RNA immunoprecipitation. A murine xenograft model was established to investigate the role of miR-182-5p in BC progression in vivo. RESULTS: An abundance of miR-182-5p was noted in BC tissues and cells. High expression of miR-182-5p was associated with poor survival. Abrogation of miR-182-5p inhibited cell proliferation and invasion in BC cells. Interestingly, PTEN was indicated as a target of miR-182-5p, and its restoration reversed miR-182-5p-mediated promotion of proliferation and invasion of BC cells. Moreover, depletion of miR-182-5p suppressed tumor growth via up-regulating PTEN expression in the murine xenograft model. CONCLUSION: MiR-182-5p exhaustion blocked cell proliferation and invasion by regulating PTEN expression, providing a novel therapeutic avenue for treatment of BC.

Huaier aqueous extract inhibits proliferation of human hepatoma SK-HEP-1 cells through up-regulation of autophagy / 中国中药杂志

The purpose of this study was to investigate the effect of Huaier on autophagy of human hepatoma SK-HEP-1 cells and the effect of autophagy on the proliferation of SK-HEP-1 cells. CCK-8 assay was used to evaluate the effect of Huaier on the proliferation of SK-HEP-1 cells under different concentrations and different times. Acridine orange staining was used to measure the effect of Huaier on the autolysosome formation in SK-HEP-1 cells. Immunofluorescence assay was applied to examine the effect of Huaier on the expression and distribution of autophagy marker LC3 in SK-HEP-1 cells. In addition， LC3 expression was also checked by immunoblot analysis in the presence of Huaier. At last， the effects of Huaier in combination with autophagy inhibitor bafilomycin A1 on the proliferation of SK-HEP-1 cells was detected by CCK-8 assay. The results showed that Huaier aqueous extract significantly inhibited the proliferation of human hepatoma SK-HEP-1 cells in a dose- and time-dependent manner. Huaier aqueous extract dramatically promoted the formation of autolysosome in SK-HEP-1 cells. Moreover， Huaier markedly increased the number and intensity of intracellular LC3 fluorescent puncta and up-regulated LC3-Ⅱ expression. These data indicated that Huaier evidently activated autophagy of SK-HEP-1 cells. Additionally， autophagy inhibition significantly attenuated the sensitivity of SK-HEP-1 cells to Huaier treatment. Therefore， autophagy activation is involved in the inhibitory effects of Huaier on the proliferation of human hepatoma SK-HEP-1 cells.

Ki-67 expression affects the clinical therapeutic effect on advanced lung adenocarcinoma / 医学研究生学报

Objective The expression level of the Ki-67 protein is a marker of the poor prognosis in non-small-cell lung cancer. This study explored the correlation of the Ki-67 expression level with the clinical efficacy in the treatment of advanced lung adenocarcinoma.Methods From January 2015 to December 2015, 92 patients with stage Ⅳ lung adenocarcinoma were treated in the Department of Respiratory and Critical Care Medicine of Nanjing General Hospital, 65 with oral gefitinib for EGFR mutation-positive or crizotinib for ALK positive (the targeted therapy \[TT\] group) and the other 27 with pemetrexed + cisplatin or nedaplatin (the chemotherapy \[CT\] group). The expression of Ki-67 was determined by immunohistochemistry, its relationship with the clinicopathological features and therapeutic effects was analyzed, and the progression-free survival (PFS) was calculated.Results Ki-67 was expressed lowly in 43 (46.7%) and highly in 49 (53.3%) of the cases. The median PFS after treatment was significantly longer in the patients with a low than in those with a high Ki-67 expression in the TT group (10 vs 7 mo, P<0.05) and in the CT group as well (8 vs 7 mo, P<0.05). Ki-67 (HR=1.011, 95% CI: 1.000-1.023) and ALK (HR=0.325, 95% CI: 0.112-0.942) were shown to be predictive factors of poor PFS.Conclusion The expression level of Ki-67 affects the effect of clinical treatment, especially that of the first-line targeted therapy, on advanced lung adenocarcinoma.

Optimization of combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile / 中成药

AIM To optimize the combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile Lindl..METHODS With enzyme consumption,enzymolysis temperature,enzymolysis time and solidliquid ratio as influencing factors,contents of dendrobine,total alkaloids and polysaccharides as evaluation indices,orthogonal test was applied to optimizing the combined enzymatic extraction.RESULTS The optimal conditions for papain extraction were determined to be 0.10 g for enzyme consumption,45 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶ 50 for solid-liquid ratio,the contents of dendrobine,total alkaloids and polysaccharides were 3.495 5,4.341 8 and 35.898 7 mg/g,respectively.The optimal conditions for cellulase extraction were determined to be 0.30 g for enzyme consumption,50 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶40 for solid-liquid ratio,the contents of three constituents were 3.514 8,4.351 3 and 36.331 2 mg/g,respectively.The optimal conditions for pectinase extraction were determined to be 0.45 g for enzyme consumption,55 ℃ for enzymolysis temperature,2.5 h for enzymolysis time,and 1 ∶ 40 for solid-liquid ratio,the contents of three constituents were 3.524 4,4.452 8 and 26.324 2 mg/g,respectively.CONCLUSION This stable and reliable method can be used for the rapid combined enzymatic extraction for alkaloids and polysaccharides from D.nobile.

Protective effects of glycyrrhizic acid against lupus nephritis in MRL/lpr mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate protective effect of glycyrrhizic acid (GA) against lupus nephritis in MRL/lpr mice and explore its underlying mechanisms.</p><p><b>METHODS</b>Forty MRL/lpr mice were randomized equally into blank control group, dexamethasone (1.5 mg/kg) group, GA (20 mg/kg) group, and GA (40 mg/kg) group with corresponding treatments for 7 days, with 10 wild-type mice as the control group. Serum levels of uric acid and creatinine and inflammatory cytokines in the serum and kidney were tested after the treatments using enzyme-linked immunosorbent assays (ELISA). The pathological changes in the kidneys were detected using HE staining, and the protein expressions of NLRP3, ASC, caspase-1, IL-1β, p-NF-κB, NF-κB, p-IκBα, and IκBα were detected with Western blotting.</p><p><b>RESULTS</b>GA obviously decreased serum levels of uric acid and creatinine, decreased inflammatory cytokines in the serum and kidney, ameliorated renal pathologies and inhibited the expressions of NLRP3, ASC, caspase-1, IL-1β, p-NF-κB, and p-IκBα proteins in MRL/lpr mice.</p><p><b>CONCLUSION</b>GA has protective effects against lupus nephritis in MRL/lpr mice.</p>

Analysis of the Hierarchical Diagnosis and Treatment System in China Based on Game Theory and the Principle of Incentive Compatibility / 中国医院管理

Objective To understand the interests of the government,large general hospitals,higher medical institutions and primary health care institutions to promote the establishment of hierarchical diagnosis and treatment system.Methods The game theory is used to analyze the contradictions and interests of the stakeholders.Results The lack of government investment,the higher medical institutions not wanting to sink medical resources,lack of capacity for primary health care institutions and other issues hinder the smooth implementation of the classification system.Conclusion The role of government should be played,and government investment be increased;the division of labor among medical institutions be strengthened,and medical resources be reasonably allocated;and the strength of primary health care institutions be improved.

Effect of diallyl disulfide on learning and memory abilities and hippocampal synapses in mouse models of Alzheimer's disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of diallyl disulfide (DADS) on hippocampal synapses and learning and memory abilities in a mouse model of A1zheimer's disease (AD).</p><p><b>METHODS</b>Mouse models of AD established by agglutinated Aβ1-42 injection in the lateral cerebral ventricle were randomized into 4 groups and treated with DADS at the daily doses of 0, 10, 50 and 100 mg/kg by gavage for 30 consecutive days. The learning and memory abilities of the mice were assessed with Morris water maze test; the structures of the dendritic spines and synapses in CA1 region of the hippocampus were observed under transmission electron microscope with silver staining; PSD95 and SYP protein and mRNA expressions in the hippocampus were detected with Western blotting and RT-PCR.</p><p><b>RESULTS</b>Compared with the AD model mice, the mice treated with 50 and 100 mg/kg DADS showed enhanced learning and memory abilities in Morris water maze test. The dendritic spines and synapses in CA1 region of the hippocampus increased obviously and hippocampal expressions of PSD95 and SYP were enhanced in mice treated with 50 and 100 mg/kg DADS.</p><p><b>CONCLUSION</b>DADS at the daily doses of 50 and 100 mg/kg can improve the learning and memory abilities and increase the number of dendritic spines and synapses in the hippocampus in mouse models of AD.</p>

Influence factors of salt-sensitive hypertension and responses of blood pressure and urinary sodium and potassium excretion to acute oral saline loading among essential hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the influence factors of salt-sensitive hypertension and to observe changes of blood pressures and urinary sodium and potassium excretion in response to acute oral saline loading among essential hypertensive patients in China.</p><p><b>METHODS</b>Essential hypertensive patients from Beijing Jinzhan second community were included in this study. Salt-sensitivity was determined via the improved Sullivan's acute oral saline loading and furosemide volume-depletion tests. Binary logistic regression analysis was applied to explore influence factors of salt-sensitive hypertension. Acute oral saline loading induced changes on blood pressures and urinary sodium and potassium excretion were observed.</p><p><b>RESULTS</b>Sixty-three salt-sensitive hypertensive patients were classified out of a total of 342(18.4%) essential hypertensive patients. Salt-sensitive patients were elder than the non-salt-sensitive patients (P < 0.05) . Binary logistic regression analysis showed that age (OR = 1.744, 95%CI:0.922-3.300, P > 0.05) , gender (OR = 0.728, 95%CI:0.374-1.415, P > 0.05) , total cholesterol level (OR = 1.168, 95%CI:0.882-1.547, P > 0.05) and 24-hour urinary sodium (OR = 0.998, 95%CI:0.995-1.002, P > 0.05) were not influencing factors of salt-sensitivity among essential hypertensive patients. Bivariate general linear models for repeated measures showed that there were significant statistical differences on blood pressures and urinary electrolytes concentrations between the beginning of trials, 2 hours after acute saline loading and 2 hours after furosemide volume-depletion(all P < 0.01). There was a greater blood pressures change in salt-sensitive patients than in non-salt-sensitive patients(all P < 0.01) while urinary electrolytes concentrations change was similar between two groups(all P > 0.05).</p><p><b>CONCLUSIONS</b>Age, gender, total cholesterol level and 24-hour urinary sodium are not influencing factors of salt-sensitivity among essential hypertensive patients in this study. Impaired pressure natriuresis during acute oral saline loading and furosemide volume-depletion tests is presented in salt-sensitive essential hypertensive patients.</p>

The effect of the magnesium supplementation on vascular calcification in rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the role of magnesium sulfate in vascular calcification, to explore the role and the mechanism of magnesium sulfate in vascular calcification.</p><p><b>METHODS</b>The vascular calcification model was established by administration of vitamin D3 plus nicotine (VDN) in SD rats. To estimate the extent of calcification by Von Kossa staining, calcium content and alkaline phosphatase activity, osteopontin (OPN) mRNA were determined by using semi-quantitative reverse-transcription polymerase chain reaction.The malondialdehyde (MDA) and nitric oxide (NO) content and activities of superoxide dismutase(SOD) were measured by biochemistry.</p><p><b>RESULTS</b>A strong positive staining of black/brown areas among the elastic fibers of the medial layer in calcified aorta by Von Kossa staining, calcium content and ALP activity in calcified arteries increased by 3.9-and 3.4-fold as compared with the controls. The expression of OPN mRNA was up-regulated by 40% (P < 0.01). The lipid peroxidation products MDA in vascular were increased 2.0-fold (P < 0.01). The NO content and SOD activity were greatly decreased by 64% and 72% (P < 0.01), respectively, compared with controls. However, calcium content and ALP activity in VDN plus magnesium sulfate group were lower than those in VDN group. Low and high dosage magnesium sulfate obviously relieved degree of calcification in the cardiovascular tissues in a dosage-dependent manner (P < 0.01).</p><p><b>CONCLUSION</b>Magnesium sulfate plays a role in the pathogenesis of vascular calcification by reducing vascular calcification and decreasing vascular injury.</p>

Epidemiological study on human echinococcosis in Hobukesar Mongolian Autonomous County of Xinjiang / 中国地方病学杂志

Objective To investigate the characteristics and distribution of human eehinococcosis in Hobukesar Mongolian Autonomous County (HMAC) in Xinjiang. Methods Using cluster sampling methods, the 2 counties (Tiebukenwusa and Narenhebuke) in HMAC were chosen as focusing areas for investigation. A survey of human echinococcosis including questionnaire, serological test and abdominal ultrasonic scan was carried out. Results The prevalence of human echinococcosis was 9.0% (64/712) by ultrasound and surgical history, including 8.7% (62/712) for cystic eehinococcosis(CE), 0.3%(2/712) for alveolar echinococcosis(AE) and 15.6%(111/712) for total of serological positives in HMAC. CE prevalence rate of different occupations, age, family slaughtering livestock and drinking water source had significant differences(P<0.05). Herdsmen as the highest risk group showed a CE prevalence of the 13.4% (27/201) in comparison with other occupations. The ages between 20 to<40 year-old were at the highest risk stage with 12.8% incidence. But CE prevalence rate of different gender, ethnic and education groups had not significant differences(P>0.05). Conclusions HMAC could be considered as a high endemic human CE region in Xinjiang. The current study reported the main risk factors may include occupations, age difference and drinking water source.

PCR Amplification of 16S-23S rDNA Intergenic Spacer Regions of Bacteria in Trauma Infection by Universal Primers / 中华医院感染学杂志

OBJECTIVE To examine the feasibility of PCR amplification of 16S-23S rDNA intergenic spacer regions of bacteria in trauma infection by a pair of universal primers for gene diagnosis.METHODS The universal primers were designed at conserved regions of the 3' end of 16S rDNA and the 5' end of 23S rDNA.Bacterial genomic DNA from selected five commom bacteria in trauma infection were amplified by PCR.PCR products were examined using electrophoresis in agarose gel,and futher analyzed by sequencing.RESULTS The PCR products were similar to that we expected on the gel,which were confirmed by the results of sequencing and alignment.CONCLUSIONS Using the universal primers,16S-23S rDNA intergenic spacer regions of bacteria in trauma infection could be amplified by PCR,which lays a solid foundation for gene diagnosis in farther studies.

Megakaryocytic dysplasia and leukemia associated phenotype in elderly patients with acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to investigate the megakaryocytic dysplasia and leukemia-associated phenotypes (LAP) of acute myeloid leukemia (AML) in the elderly. The megakaryocytic dysplasia, lineage infidelity, asynchronous antigen expression, total WBC count, and karyotypes were observed in the 147 none M(3)-AML patients. Logistic regression were used to analyzed the difference between the elderly (age > or = 60) and the control. The results showed that out of the total 147 patients (66 elderly patients, and 81 younger patients) 124 patients accepted induction chemotherapy, in which 70 cases achieved complete remission (elderly 18, younger 52, p = 0.008); megakaryocytic dysplasia was found in 32 patients (21.8%); CD33 and CD19/CD7 (lineage infidelity) was co-expressed in 55 patients (37.4%), CD34 and CD11b (asynchronous antigen expression) was co-expressed in 65 patients (44.2%); white blood cell count > 25 x 10(9)/L was found in 52 patients (35.4%). By the Logistic regression, compared with the control, in the elderly patients there was difference in the megakaryocytic dysplasia, and the co-expression of CD33/CD19/CD7 and CD34/CD11b (OR = 4.315, 2.761, 0.397; p = 0.001, 0.006, 0.020), but there was no difference in the total WBC count and karyotypes (OR = 0.802, 1.096; p = 0.646, 0.813). It is concluded that the incidence of megakaryocytic dysplasia, such as lineage infidelity, and asynchronous antigen expression, in elderly patients is higher than that in younger patients.

The effect of angiotensin-converting enzyme inhibitors and aldosterone receptor blockers on cardiac function in calcium-overload rats / 中国应用生理学杂志

<p><b>AIM</b>To observe the effect of angiotensin-converting enzyme inhibitors (ACEI) and aldosterone receptor blockers on cardiac function to explore the mechanism of cardiac function descending and myocardial injury in calcium-overload rats.</p><p><b>METHODS</b>Calcium-overload in rat was induced by administration of Vitamin D3 plus nicotine. To Estimate the extent of calcium-overload by calcium content. Angiotension II and aldosterone levels in the myocardia were measured by radioimmunoassay. Cardiac function (+/- LVdp/dt, LVESP and LVEDP) were measured by Powerlab. The malondialdehyde (MDA) content, activities of lactate dehydrogenase (LDH) and creatine kinase (CPK) were measured by biochemistry.</p><p><b>RESULTS</b>Calcium content increased by 3.2-, 5.8 -fold in myocardial and artery, compared with controls. VDN-treated survivors showed lower + LVdp/dt(max) and -LVdp/dt(max) values, by 27% and 34%, respectively (both P < 0.01). Higher LVESP, and LVEDP by 42 % and 32% (P < 0.01); heart rate and mean arterial pressure were not significantly altered (P > 0.05). The lipid peroxidation products MDA and conjugated diene in myocardia were increased 22% (P < 0.01), 68% (P < 0.05) (P < 0.05), respectively. The plasma activity of CPK and LDH was greatly increased by 4.5-and 3.1-fold (P < 0.01), respectively. ACEI and spironolactone obviously relieved degree of calcium-overload and improved cardiac function and myocardial injury(P < 0.01). Calcium content in myocardia and artery was lower 44%, 39% and 57%, 34%. Lower MDA by 20%, 30%, lower conjugated diene by 44%, 35% than calcium-overload group. The plasma activity of CPK and LDH were obviously decreased 28%, 34% and 20%, 27%, compared with calcium-overload group.</p><p><b>CONCLUSION</b>Calcium-overload could lead to cardiac function descending and myocardial injury in calcium-overload rats by VDN. ACEI and spironolactone could reduce calcium-overload in myocardial and ameliorate cardiac function and decrease myocardial injury.</p>

The leakage radiation testing method for mammographic X-ray equipments / 中国医疗器械杂志

This article introduces a new testing method for mammographic x-ray equipments. The films placed around the x-ray tube assembly are exposured to find the location of leakage radiation and then the accurate testing for the leakage radiation of the mammographic x-ray equipment is carried out with a radiation dosimeter.

Real-time fluorescent quantitative PCR for detection of peripheral blood T-cell receptor excision circles / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop and optimize real-time fluorescence quantitative PCR (FQ-PCR) with the housekeeping gene RAG2 as cell number control to quantify T-cell receptor excision circle (TREC) in the peripheral blood.</p><p><b>METHODS</b>The real-time PCR system for amplifying TREC and RAG2 genes was established on the basis of ABI 7000 apparatus using Golden Taq system, designed primers, TaqMan-MGB probes and optimized buffer. PCR conditions were optimized with standard samples of TREC plasmid.</p><p><b>RESULTS</b>The amplification with the primer pair T(3) and T(4) was more efficient than that with T(1) and T(2). More specific and efficient amplification in FQ-PCR was achieved using TaqMan-MGB probes as compared with general Taq-Man probes. Golden Taq was more effective than general Taq in improving the specificity and decreasing the artifact, and 95 degrees C; for 10 min, 95 degrees C; for 5 s, and 53 degrees C; for 30 s for a total of 40 cycles using ABI7000 was found as the optimized thermal parameter setting.</p><p><b>CONCLUSION</b>An optimized real-time PCR protocol for detecting TREC in peripheral blood mononuclear cells is established.</p>

Knockdown of bcl-xL expression with RNA interference induces nasopharyngeal carcinoma cells apoptosis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the inhibition of the expression of bcl-xL gene induced by RNA interference in CNE-2Z cell line in addition to the inhibition of its proliferation and apoptotic induction.</p><p><b>METHODS</b>Small interfering RNAs targeting bcl-xL gene were synthesized by using web design software provided by Amnion and the silencer short interfering RNA (siRNA) construction kit; fluorescein-labeled siRNAs were done by FAM-silencer siRNA labeling kit; siRNAs were transfected into CNE-2Z cells by using lipofectamine 2000 reagent; siRNA transfection efficiencies were analyzed by fluorescent microscopy; down-regulation of bcl-xL was detected by RT-PCR; thiazolyl blue (MTT) assay was used to assess the cell growth; apoptosis of CNE-2Z cells was analyzed by flow cytometry.</p><p><b>RESULTS</b>Green fluorescence in the cells was seen clearly in FAM-labeled siRNA transfected group under the fluorescent microscope while none in the untransfected group. Different down-regulations of bcl-xL mRNA expression were found in the transfected groups. The expression of bcl-xL mRNA decreased by 10% - 70% in the siRNAs transfected CNE-2Z by RT-PCR scan analysis. The inhibitory rate of cell proliferation depended on time and concentrations to some extent. Different cell apoptosis could be induced by different concentrations of siRNA4.</p><p><b>CONCLUSIONS</b>The synthesized siRNAs in vitro were able to down-regulate the expression of bcl-xL There were different capabilities of the specific siRNAs down-regulation. The transient transfected bcl-xL siRNA4 could effectively inhibit the growth of the cancer cells and induce theirs apoptosis. It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for anti-nasopharyngeal carcinoma gene therapy.</p>

Construction of a lentiviral expression vector that inhibits enhanced green fluorescent proteins / 医学研究生学报

Objective: To construct a lentiviral expression vector that inhibits enhanced green fluorescent proteins(EGFP).Methods: The EGFP shRNA was cloned into the pLL3.7 lentivirus vector digested with XbaⅠ/XhoⅠ.J558L and Hela cells were infected by lentiviruses,and the EGFP expression was observed with the fluorescence microscope.Results: The EGFP RNAi sequence was correctly cloned into the lentivirus vector with the U6 promoter,and the EGFP expressions of Hela and J558L cells were suppressed after infected by lentivirus containing EGFP shRNA. Conclusion: The lentivirus vector effectively inhibited EGFP expression,which could be used to mediate further researches on gene function.