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Biomedical subjects

Lei Li

Publications and source records attributed to Lei Li.

At least 145 records · Page 8Linked to original sources

Circadian rhythms of behavioral cone sensitivity and long wavelength opsin mRNA expression: a correlation study in zebrafish.

Using a behavioral assay based on visually mediated escape responses, we measured long-wavelength-sensitive red cone (LC) sensitivities in zebrafish. In a 24 h period, the zebrafish were least sensitive to red light in the early morning and most sensitive in the late afternoon. To investigate if the fluctuation of behavioral cone sensitivity correlates with opsin gene expression, we measured LC opsin mRNA expression at different times in the day and night under different lighting conditions. Under a normal light-dark cycle, the expression of LC opsin mRNA determined by real-time RT-PCR was low in the early morning and high in the late afternoon, similar to the fluctuation of behavioral cone sensitivity. This rhythm of LC opsin mRNA expression, however, dampened out gradually in constant conditions. After 24 h of constant light (LL), the expression of LC opsin mRNA dropped to levels similar to those determined in the early morning in control animals. By contrast, when the zebrafish were kept in constant dark (DD), the expression of LC opsin mRNA increased, to levels about 30-fold higher than the expression in the early morning in control animals. This day-night fluctuation in LC opsin mRNA expression was correlated to changes in opsin density in the outer segment of cone photoreceptor cells. Microspectrophotometry (MSP) measurements found significant differences in red cone outer segment optical density with a rhythm following the behavioral sensitivity. Furthermore, dopamine modulated the circadian rhythms in expression of LC opsin mRNA. Administration of dopamine increased LC opsin mRNA expression, but only in the early morning.

Animals↗

Improving P-gp expression in human mononuclear cells in vitro transfected by multidrug resistance-1 mRNA.

OBJECTIVE: To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1 (MDR1) mRNA. METHODS: Two MDR1 cDNA vectors, pT7TS_MDR1 and pGEM5Zf(+)_MDR1, were constructed and transcripted in vitro. Vector pGEM5Zf(+)_MDR1 only contained the coding region of mdr1 cDNA, and pT7TS_MDR1 also included Xeponus beta-globin 5' and 3' untranslated region. MNCs were prepared from peripheral blood of parvicellular lung cancer patient. The two human mdrl mRNAs were then transferred into human MNCs in vitro by DOTAP. And the expression efficiency and pump function of P-gp were measured with flow cytometry. RESULTS: Expression of P-gp significantly elevated in both transferred cells compared with untransferred cells (P < 0.01). And pT7TS_MDR1 showed higher capability in elevating the expression of P-gp than pGEM5Zf(+)_MDR1 (P < 0.01). The P-gp function was elevated in both pT7TS_MDR1 and pGEM5Zf(+) MDR1 groups. The survival ratio of MNCs in erythrocyte-lysis-solution (ELS, 86.07%) and lymphocyte-isolation-solution (LIS, 83.67%) had no significant difference. The CD34+ cells content of the MNCs used for transfection was 2.65% and 1.01% in ELS and LIS group, respectively (P < 0.01). CONCLUSIONS: It is a feasible approach to improve P-gp expression in human MNCs by transfection of MDR-1 mRNA. And the ELS may be more suitable for purifing MNCs for mRNA transfection than LIS.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

[Primary study on the method for separating mature sperms from human testicular tissue].

This study was aimed to explore a simple and applicable method of separating mature sperms from human testicular tissue for intracytoplasmic sperm injection and embryo transfer. The suspension of human testicular tissue was cultured in 10% human serum albumin and human tubule fluid with different concentrations (0 u/ml; 50 u/ml; 100 u/ml; 150 u/ml; 200 u/ml) of hyaluronidase for 24 h, and then the Percoll gradient centrifugation was processed to separate the sperms; meanwhile the sperms were counted and graded according to their motility. The difference in quality and quantity among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treated groups contained large quantity and high quality of sperms as compared with the two contrast groups (P<0.01). The groups in the solution of 50 u/ml, 100 u/ml and 150 u/ml concentrations of hyaluronidase had almost the same amount of sperms that displayed higher motility as compared against the sperms in the group treated with 200 u/ml concentration of hyaluronidase (P<0.01). There was no difference between the two contrast groups (P>0.05), or among the groups treated with 50 u/ml, 100 u/ml, and 150 u/ml of hyaluronidase concentration (P>0.05). This method of adopting hyaluronidase with Percoll gradient centrifugation in the process for separating mature sperms from human testicular tissue is applicable. It can increase the quantity and quality of sperms separated from testicular tissue suspensions when adequate concentrations of hyaluronidase is used.

Adult↗

[Heterozygous mutation in KCNQ1 cause Jervell and Lange-Nielsen syndrome].

OBJECTIVE: Jervell and Lange-Nielsen syndrome (JLNS) is a severe cardioauditory syndrome manifested as QT interval prolongation, abnormal T waves, and relative bradycardia ventricular tachyarrhythmias. In this report, we screened a nonconsanguineous families with JLNS for mutations in KCNQ1. METHODS: Mutation analysis was performed by using purified PCR products to direct sequence analysis on an ABI-3730XL automated DNA sequencer. The whole sequence of proband' KCNQ1 was screened firstly, then screened the mutation exon sequences of others of the family and 50 unrelated normal persons. RESULTS: A heterogeneous mutation was identified in the patients of the JLNS family, a missense mutation (G-->T) at nucleotide 917 encoded in exon 6 of KCNQ1. This substitution leads to a change from glycine to Valine at codon 306(G306V) corresponding to the S5 transmembrane segment of KCNQ1. The other normal members of the family and 50 unrelated normal persons were not identified this mutation. CONCLUSION: The result suggested that not only homozygous mutations or compound heterozygous mutations in KCNQ1 could cause Jervell-Lange-Nielsen syndrome, the single heterozygous mutation may also cause Jervell-Lange-Nielsen syndrome.

Adolescent↗

[Effects of neural stem cells transplantation on glial cell line-derived neurotrophic factor and growth associated protein 43 after spinal cord injury in rats].

OBJECTIVE: To observe the effects of neural stem cells(NSCs) transplantation on the glial cell line-derived neurotrophic factor (GDNF) and growth associated protein 43(GAP-43) after the spinal cord injury (SCI), and to investigate the mechanism of repairing the SCI by NSCs transplantation. METHODS: The neural stem cells from the hippocampus of rats' embryo were cultured and identified by immunocytochemistry. The SCI model was made by the modified Allen device. Sixty adult Wistar rats were randomly divided into three groups: spinal cord injury was treated with transplantation of NSCs (group A, n = 24), with DMEM solution (group B, n = 24) and normal control group without being injured(group C, n = 12). Seven days after the operation of SCI, the NSCs were transplanted into the injured site. Then GAP-43 and GDNF expressions were tested by RT-PCR and immunohistochemistry. RESULTS: Compared with group B, the GDNF mRNA expression of group A increased by 23.3% on the 1st day, by 26.8% on the 3rd day and by 32.7% on the 7th day; the GAP-43 mRNA expression increased by 19.5% on the 1st day, 21.6% on the 3rd day and 23.1% on the 7th day. There were statistically significant differences (P<0.05). CONCLUSION: The transplantation of NSCs can change the micro-environment injured site and promote the regeneration of axon by enhancing the expressions of GDNF mRNA and GAP-43 mRNA. It is one of the mechanisms of repairing the SCI by NSCs transplantation.

Animals↗

[Evaluation of the "Jiangsu/WHO 100% condom use programme in China"].

OBJECTIVE: To evaluate the effect and to explore the feasible model of 100% condom use programme (100%CUP) under existing policy and law milieu, so as to provide information for the adjustment of HIV/AIDS policy and intervention strategies. METHODS: Information on knowledge, attitude and behavior of female sex workers (FSWs) in entertainment establishments were collected and followed by qualitative and quantitative analysis including: samples tested for Chlamydia; a repeated cross-investigation and the final evaluation on the effect of the programme. RESULTS: The reported rate of condom use at the last sex activity with clients increased from 74.9% to 92.2% while the rate of sexually transmitted disease (STD) prevalence droped from 25.1% to 14.1% after three year's intervention program. The number of condoms distributed increased from 50,000 per quarter of a year in 2001 to over 100,000 in 2003. The rates of awareness on the knowledge of STD/AIDS prevention increased obviously. CONCLUSION: The 100%CUP was proved to be feasible and effective in the pilot site, and should be expanded in entertainment establishments.

Adult↗

[Detecting the API2-MALT1 fusion gene and its significance in Hashimoto's thyroiditis and thyroid lymphomas].

OBJECTIVE: To investigate the expression of API2-MALT1 fusion gene mRNA in Hashimoto's thyroiditis, thyroid MALT lymphoma and DLBCL, as well as the correlation between the expression and the pathogenesis and prognosis of two types of thyroid lymphoma. METHODS: Eight cases of Hashimoto's thyroiditis, 12 cases of MALT lymphoma and 3 cases of DLBCL in thyroid were selected and retrieved. API2-MALT1 transcripts were investigated in all cases by reverse transcript touchdown-PCR and nested PCR. The effect of API2-MALT1 fusion gene on the prognosis of thyroid lymphomas was assessed by analyzing the clinicopathological features and the follow-up findings. RESULTS: The variant 1446/1123 transcript was detected in 2 of thyroid lymphomas (1 MALT lymphoma, 1 DLBCL) but not in 8 cases of Hashimoto's thyroiditis and 5 cases of chronic lymphadenitis. Both of API2-MALT1-positive cases were accompanied with Hashimoto's thyroiditis, showing stage I E and no involvement of lymph node. CONCLUSION: The incidence of API2-MALT1 transcript in thyroid lymphomas was low; however, the correlation between API2-MALT1 fusion gene and the progression from Hashimoto's thyroiditis to lymphoma could not be excluded; the thyroid lymphoma with API2-MALT1 transcript seemed to develop on a more indolent clinical course.

Adult↗

[Expression of bcl-10 protein in MALT lymphoma].

OBJECTIVE: To investigate the significance of bcl-10 protein expression in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) lymphoma. METHODS: Sixty-two cases of MALT lymphoma were reviewed and immunohistochemical studies for bcl-10 and Ki-67 were performed. RESULTS: Sixty out of the 62 cases studied (96.8%) were positive for bcl-10. Thirty-three (53.2%) showed bcl-10 expression in both the nuclei and cytoplasm, while 27 cases (43.6%) showed only cytoplasmic staining. The 10 cases with Hashimoto's thyroiditis demonstrated bcl-10 expression in the cytoplasm. The mean age of patients with bcl-10 nuclear expression (51.4 years old) was 5.2 years younger than those (56.6 years) without bcl-10 nuclear expression. The former category also showed a male predilection (male to female ratio = 19:14, in contrast to 10:19 in the latter category). The frequency of bcl-10 nuclear expression was lower in cases from thyroid but higher in cases from lung, stomach and intestine (P < 0.05). There was no statistically significant correlation between bcl-10 nuclear expression and clinical tumor stage (P > 0.05) or tumor cell morphology (P > 0.05). Amongst the 40 cases of gastrointestinal MALT lymphoma, bcl-10 nuclear expression correlated with extent of tumor involvement. The protein was expressed in 36.4% (4 out of 11 cases) of MALT lymphoma confined to mucosa or submucosa, 65.2% (15 out of 23 cases) of those invading down to muscularis propria or subserosa, and 100% (all 6 cases) of those extending beyond serosa (P < 0.05). There was no statistically significant difference in Ki-67 proliferative index between bcl-10-positive and bcl-10-negative groups (P < 0.05). Follow-up data were available in 52 patients (83.9%) and the five-year survival rate was no statistically significant difference in survival between bcl-10-positive (29 patients, 96.3%) and bcl-10-negative groups (23 patients, 66.4%, P > 0.05). CONCLUSIONS: Two expression patterns of bcl-10 protein were observed in MALT lymphoma: mixed nuclear-cytoplasmic and cytoplasmic only. The bcl-10 nuclear expression appears more important and correlates with anatomic site of tumor and extent of tumor involvement. Immunohistochemical detection of bcl-10 may carry some diagnostic and prognostic implications in assessment of MALT lymphoma.

Adaptor Proteins, Signal Transducing↗

Synthesis, structure, spectroscopic properties, and electrochemistry of (1,8,15,22-tetrasubstituted phthalocyaninato)lead complexes.

Three (1,8,15,22-tetrasubstituted phthalocyaninato)lead complexes Pb[Pc(alpha-OR)(4)] [H(2)Pc(alpha-OC(5)H(11))(4) = 1,8,15,22-tetrakis(3-pentyloxy)phthalocyanine; H(2)Pc(alpha-OC(7)H(15))(4) = 1,8,15,22-tetrakis(2,4-dimethyl-3-pentyloxy)phthalocyanine; H(2)Pc(alpha-OC(10)H(7))(4) = 1,8,15,22-tetrakis(2-naphthyloxy)phthalocyanine] (1-3) have been prepared as racemic mixtures by treating the corresponding metal-free phthalocyanines H(2)Pc(alpha-OR)(4) (4-6) with Pb(OAc)(2).3H(2)O in refluxing n-pentanol. The molecular structure of Pb[Pc(alpha-OC(5)H(11))(4)] (1) in the solid state has been determined by single-crystal X-ray diffraction analysis. This compound, having a nonplanar structure, crystallizes in the monoclinic system with a P2(1)/c space group. Each unit cell contains two pairs of enantiomeric molecules, which are linked by weak coordination of the Pb atom of one molecule with an aza nitrogen atom and its neighboring oxygen atom from the alkoxy substituent of another molecule, forming a pseudo-double-decker supramolecular structure in the crystals with a short ring-to-ring separation, 2.726 A, and thus a strong ring-ring pi-pi interaction. The decreased molecular symmetry for these complexes has also been revealed by the NMR spectra of 1 and 2. The methyl protons of the 3-pentyloxy and 2,4-dimethyl-3-pentyloxy side chains of 1 and 2, respectively, are chemically inequivalent. In addition to the elemental analysis and various spectroscopic characterizations, these compounds have also been electrochemically studied. Two one-electron oxidations and up to five one-electron reductions have been revealed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methods.

Journal Article↗

[Mutations in beta myosin heavy chain gene: two mutations in Chinese with familial hypertrophic cardiomyopathy and the correlation between the genotype and phenotype].

OBJECTIVE: To study the disease-causing gene mutation in Chinese with hypertrophic cardiomyopathy (HCM), and to analyze the correlation between the genotype and phenotype. METHODS: Samples of peripheral blood were collected from five Chinese patients with HCM in whose families at least 2 HCM patients existed. The exon in the functional regions of the beta myosin heavy chain gene (beta-MHC) were amplified with PCR and the products were sequenced. The relation between the genotype and phenotype was analyzed. RESULTS: Two mutations were first identified. Eighty controls were normal in the genetic test. CONCLUSION: beta-MHC may be the main disease-causing gene. Two mutations have different phenotypes. In one family, the identical mutation has different phenotypes and prognoses. The heterogeneity of phenotype suggests that multiple factors be involved in the pathogenesis.

Adolescent↗

[The therapeutic effect of oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine in mice with acute necrotizing pancreatitis].

OBJECTIVE: To investigate the therapeutic effect of oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (OXPAPC) in animal model of acute necrotizing pancreatitis (ANP) and its possible mechanism. METHODS: Forty-eight C57BL/6J mice were randomly divided into two groups, ANP group and ANP treated with OXPAPC group. ANP model was induced by seven times administration of cerulein (50 microg/kg) challenged by lipopolysaccharide (LPS) (5 mg/kg) intraperitoneal injection. Treatment with OXPAPC was started 5 mins before LPS injection. At the 9th hour, 12nd hour and 24th hour after the first injection of cerulein, serum levels of amylase and lactate dehydrogenase (LDH) were measured. Severity of pancreatitis was evaluated by histological scoring system. The activity of myeloperoxidase (MPO) in pancreas was determined by zymohistochemistry. Intrapancreatic TNF-alpha, IL-1beta, ICAM-1 and E-selectin mRNA expressions in pancreas were studied by semi-quantitative RT-PCR. The activation of nuclear factor kappaB (NF-kappaB) and c-Jun-N-terminal kinase 1 (JNK1) were investigated by the methods of western blot. RESULTS: Treatment with OXPAPC significantly decreased serum amylase and LDH levels at the 9th hr and 12nd hour. Histologically, OXPAPC reduced the severity of pancreatic injury including inflammatory cell infiltration and necrosis at 12nd hour and 24th hour. There was a significant decrease of MPO activity in OXPAPC group as compared to ANP group. Intrapancreatic TNF-alpha, IL-1beta, ICAM-1 and E-selectin mRNA levels were reduced in OXPAPC-treated group. Western blot showed down-regulation of NF-KappaB p65 and JNK1 expression were found in OXPAPC-treated mice. CONCLUSION: OXPAPC decreased the severity of experimental acute pancreatitis in mice and inhibited the intrapancreatic inflammatory mediators expression in vivo. The protective effect of OXPAPC was mediated, at least in part, through blocking endotoxin signal pathway.

Amylases↗

Disruption of the Rad9/Rad1/Hus1 (9-1-1) complex leads to checkpoint signaling and replication defects.

The checkpoint sliding-clamp complex, Rad9/Rad1/Hus1, plays a critical role during initiation of checkpoint signals in response to DNA damage and replication disruption. We investigated the impact of loss of Rad1 on checkpoint function and on DNA replication in mammalian cells. We show that RAD1 is an essential gene for sustained cell proliferation and that loss of Rad1 causes destabilization of Rad9 and Hus1 and consequently disintegration of the sliding-clamp complex. In Rad1-depleted cells, Atr-dependent Chk1 activation was impaired whereas Atm-mediated Chk2 activation was unaffected, suggesting that the sliding clamp is required primarily in Atr-dependent signal activation. Disruption of sliding-clamp function also caused a major defect in S-phase control. Rad1-depleted cells exhibited an RDS phenotype, indicating that damage-induced S-phase arrest was compromised by Rad1 loss. Furthermore, lack of Rad1 also affected the efficiency of replication recovery from DNA synthesis blockage, resulting in a prolonged S phase. These deficiencies may perpetually generate DNA strand breakage as we have found chromosomal abnormalities in Rad1-depleted cells. We conclude that the Rad9/Rad1/Hus1 complex is essential for Atr-dependent checkpoint signaling, which may play critical roles in the facilitation of DNA replication and in the maintenance of genomic integrity.

Cell Cycle↗

Tetrabromobutatriene: completing the perhalocumulene series.

[structure: see text] Tetrabromobutatriene, C(4)Br(4), can be prepared directly from dibromobutadiyne by reaction with Br(2) at -25 degrees C in concentrated hexanes solution. The cumulene precipitates out of the reaction mixture as a yellow powder. Under palladium-catalyzed coupling conditions, C(4)Br(4) can undergo allylic rearrangement, giving a mixture of products, including some with butenyne backbones. However, in furan solution, C(4)Br(4) reacts cleanly at its central double bond to give the furan Diels-Alder adduct. Under Suzuki conditions, this adduct reacts at the furan double bond rather than at bromide.

Journal Article↗

Cooperative molecular recognition of dyes by dyad and triad cyclodextrin-crown ether conjugates.

Three beta-cyclodextrin (beta-CyD) derivatives with crown ether units, that is N-(4'-benzo-15-crown-5)-6-imino-6-deoxy-beta-CyD (2), 6,6'-[N-(4,4'-dibenzo-18-crown-6)-imino]-bridged bis(beta-CyD)(3), and 2,2'-[O-(4',5'-benzo-15-crown-5)-ethyl]-bridged bis (beta-CyD)(5), were synthesized as cooperative recognition receptor models. Their molecular binding behavior with four representative fluorescent dyes, i.e., ammonium 8-anilino-1-naphthalenesulfonate (ANS), sodium-6-toluidino-2-naphthalene-sulfonate (TNS), Acridine Red (AR) and Rhodamine B (RhB), was investigated in buffer solutions (pH = 7.20) at 25 degreesC by means of circular dichroism, NMR and fluorescence spectroscopy. 2D-ROESY experiments showed that dyad host 2 and triad host 3 adopted a CyD-guest-crown ether binding mode, while triad host 5 adopted a CyD-guest-CyD binding mode, upon inclusion complexation with guest molecules. Therefore, hosts 2 and 3 showed high molecular recognition ability towards charged guests, giving an enhanced binding ability up to 115 times for ANS by 3 and fairly high molecular selectivity up to 1450 times for the ANS/AR pair by 2 as compared with native beta-CyD in an aqueous phosphate buffer solution. On the other hand, host 5 was found to be able to effectively recognize the shape of a guest molecule, showing significantly higher binding ability towards linear guests. The binding affinities and molecular recognition abilities of these CyD-crown ether conjugates towards guest molecules are discussed from the viewpoint of electrostatic and/or hydrophobic interactions, size/shape-fit concept, and multiple recognition mechanism between host and guest.

Anilino Naphthalenesulfonates↗

Advances in the study of luminescence probes for proteins.

Spectral probes (or labels) have been widely used for the investigation and determination of proteins and have made considerable progress. Traditional luminescence probes include fluorescent derivatizing reagents, fluorescent probes and chemiluminescence probes which continue to develop. Of them, near infrared (NIR) fluorescent probes are especially suitable for the determination of biomolecules including proteins, so their development has been rapid. Novel luminescence probes (such as nanoparticle probes and molecular beacons) and resonance light scattering probes recently appeared in the literature. Preliminary results indicate that they possess great potential for ultrasensitive protein detection. This review summarizes recent developments of the above-mentioned probes for proteins and 195 references are cited.

Luminescent Measurements↗

Dual neuroprotective signaling mediated by downregulating two distinct phosphatase activities of PTEN.

The tumor suppressor PTEN (phosphatase and tensin homolog deleted on chromosome 10) is a lipid and protein phosphatase. We report here that PTEN physically associates with the NR1 and NR2B subunits of NMDA receptors (NMDARs) in rat hippocampus. Downregulating the protein expression of PTEN inhibits the function of extrasynaptic NMDARs and decreases NMDAR surface expression, suggesting a crucial role for endogenous PTEN in the modulation of NMDAR-mediated neuronal function. Reducing PTEN expression also enhances Akt/Bad phosphorylation in hippocampal neurons. Importantly, suppressing lipid and protein phosphatase activity of PTEN, respectively, activates Akt and inhibits extrasynaptic NMDAR activity and thereby protects against ischemic neuronal death in vitro and in vivo. Thus, our study reveals a dual neuroprotective mechanism by which Akt/Bad and extrasynaptic NMDARs are regulated via downregulation of two distinct PTEN phosphatase activities and present the possibility of PTEN as a potential therapeutic target for stroke treatment.

Animals↗

Circadian regulation of GABAA receptor function by CKI epsilon-CKI delta in the rat suprachiasmatic nuclei.

The type A GABA receptors are thought to mediate synchronization of clock cell activity within the suprachiasmatic nuclei (SCN). Here we report that casein kinases I epsilon and delta (CKI epsilon and CKI delta), the crucial clock regulators, form a complex with GABA(A) receptors and inhibit the receptors' function within the SCN according to a circadian rhythm. These results indicate that circadian variation of the kinase-receptor association may mediate regulation of GABA(A) receptor function by CKI epsilon-CKI delta in the SCN.

Analysis of Variance↗