Biomedical subjects
Publications and source records attributed to Q Wei.
Dendrobium drug, a rare medicinal herb, reproduces differcut for growth environment to lead rare resources. By studying of cultivation for many many years in Fuliu mountain area, authors had found out the cultivated method simulating the habitat of wild Dendrobium flexicaule. The results could supply a basis for its development and utilization.
Unlike wild-type mouse melanocytes, where melanosomes are concentrated in dendrites and dendritic tips, melanosomes in dilute (myosin Va-) melanocytes are concentrated in the cell center. Here we sought to define the role that myosin Va plays in melanosome transport and distribution. Actin filaments that comprise a cortical shell running the length of the dendrite were found to exhibit a random orientation, suggesting that myosin Va could drive the outward spreading of melanosomes by catalyzing random walks. In contrast to this mechanism, time lapse video microscopy revealed that melanosomes undergo rapid ( approximately 1.5 microm/s) microtubule-dependent movements to the periphery and back again. This bidirectional traffic occurs in both wild-type and dilute melanocytes, but it is more obvious in dilute melanocytes because the only melanosomes in their periphery are those undergoing this movement. While providing an efficient means to transport melanosomes to the periphery, this component does not by itself result in their net accumulation there. These observations, together with previous studies showing extensive colocalization of myosin Va and melanosomes in the actin-rich periphery, suggest a mechanism in which a myosin Va-dependent interaction of melanosomes with F-actin in the periphery prevents these organelles from returning on microtubules to the cell center, causing their distal accumulation. This "capture" model is supported by the demonstration that (a) expression of the myosin Va tail domain within wild-type cells creates a dilute-like phenotype via a process involving initial colocalization of tail domains with melanosomes in the periphery, followed by an approximately 120-min, microtubule-based redistribution of melanosomes to the cell center; (b) microtubule-dependent melanosome movement appears to be damped by myosin Va; (c) intermittent, microtubule-independent, approximately 0.14 microm/s melanosome movements are seen only in wild-type melanocytes; and (d) these movements do not drive obvious spreading of melanosomes over 90 min. We conclude that long-range, bidirectional, microtubule-dependent melanosome movements, coupled with actomyosin Va-dependent capture of melanosomes in the periphery, is the predominant mechanism responsible for the centrifugal transport and peripheral accumulation of melanosomes in mouse melanocytes. This mechanism represents an alternative to straightforward transport models when interpreting other myosin V mutant phenotypes.
To develop an animal model for mucosal HIV-1 infection, adult chimpanzees were inoculated without trauma by depositing the virus inoculum at the entrance to the cervical canal with a rigid catheter to which flexible tubing was attached. By this procedure, persistent infections were established in some chimpanzees with various infectious doses of either cell-associated HIV-1LAI(IIIB) (peripheral blood mononuclear cells from an infected chimpanzee) or with cell-free HIV-1 strains representing subtypes B and E, but not with a subtype A strain. Although some animals did not become infected until after the second or third cervicovaginal exposure, one chimpanzee was clearly infected after one exposure by several criteria, including virus isolation, but this animal did not seroconvert. A second chimpanzee appeared to be resistant to infection despite repeated mucosal exposures at irregular intervals. However, lymphocytes from both of these animals exhibited low-level proliferative responses to HIV-1 but not SIV antigens. Despite these apparently abortive or latent infections, after exposure to HIV-1 by the intravenous route, both animals developed systemic infections and seroconverted. Overall, 8 of 10 chimpanzees were infected systemically after one to three cervicovaginal exposures to HIV-1LAI(IIIB). The results indicate that (1) HIV-1 productive infection of female chimpanzees by the cervicovaginal route generally requires more than one exposure, just as with humans; (2) low level infections without seroconversion can be established after mucosal exposure to HIV; and (3) vaccine efficacy studies involving a single virus challenge of immunized chimpanzees by the cervicovaginal route probably will not be possible.
DDVP was isolated from white spirits using Solid-Phase Microextraction (SPME) technique, and determined by wide-bore capillary gas chromatography (GC) coupling a flame ionization detector. This method had an excellent linearity among 0.5-12 mg/L. Correlation coefficient and minimum detectable concentration were 0.9983 and 0.05 mg/L respectively. This method enables the extraction, the enrichment and the injection to integrate into a single step, and it was free-solvent in whole process. SPME is an advanced technique for the extraction and analysis of the samples.
The agonist-dependent activation and desensitization of a recombinant analog of delta-opioid receptor lacking the carboxyl-terminal (C-terminal) 31 residue peptide segment were discussed. The cDNA of the C-truncated delta-opioid receptor was created by polymerase chain reaction (PCR), stably expressed in Chinese hamster ovary cells and characterized by binding assay and function assay. Agonist [D-Pen2, D-Pen5]-enkephalin (DPDPE) stimulated the specific [35S]GTPgammaS binding to membrane fragments of cells expressing the C-truncated and the wild-type delta-opioid receptors in different levels dose-dependently. EC50 values of DPDPE on truncated and wild-types in stimulation were very similar. Agonist-dependent desensitization, which could be blocked by protein kinase inhibitor staurosporine (Stau), was observed after pretreating truncated receptors with 1 microM DPDPE for 10 min, the same as wild-types. The results reveal that the C-terminal of the delta-opioid receptor is not involved in controlling the G-protein activation and phosphorylation-related functional desensitization.
Class V unconventional myosins are two-headed, nonfilamentous, actin-based mechanoenzymes that appear to be expressed ubiquitously. Mice possess at least two myosin V heavy chain genes (dilute and myr6) whose approximately 190 kDa protein products are referred to as myosin Va and Vb, respectively. Using antibodies that are specific for the Va isoform and immunofluorescence microscopy, we show here that myosin Va localizes to the microtubule organizing center (MTOC) in interphase cells, and to the mitotic asters, spindle, and midbody of dividing cells. These associations, which in the case of mitotic cells are characterized by the concentration of myosin Va in the immediate vicinity of the microtubules, were observed in a variety of cell types, including primary and immortal mouse melanocytes and fibroblasts, Hela cells, and Cos cells. Importantly, these associations were not observed in melanocytes and fibroblasts cultured from dilute null mice, indicating that the staining of these microtubule-rich domains was due to the presence of myosin Va, as opposed to another protein(s) containing a shared epitope(s) with myosin Va. When cells were extracted with detergent prior to fixation, myosin Va remained associated with each of these microtubule-rich domains, suggesting that these associations are not due to the possible presence of membranes at these sites. This fact, and our observation that these microtubule-rich domains contain little if any F-actin (based on phalloidin staining), suggest that myosin Va may bind to microtubules either directly or through a microtubule-associated protein. Finally, we found that dilute null fibroblasts in primary culture are twice as likely to be binucleate as wild type fibroblasts of the same genetic background (35% vs. 17%). Together, these results indicate that myosin Va associates with microtubule-rich domains in both interphase and dividing cells, and plays a role in the efficiency of cell division in culture.
Relatively benign Theileria parasites are widespread among cattle in East Asia. Although the parasites are presumed to be of the Theileria sergenti/Theileria buffeli/Theileria orientalis group, their taxonomic status and epidemiology have not been well defined. In the present study, theilerial DNA samples were collected from various East Asian countries, including Japan, Korea, Taiwan, and China. DNA sequences encoding a major piroplasm surface protein were amplified by polymerase chain reaction, followed by cloning into a plasmid vector. More than 20 DNA clones derived from parasite DNA of a single infected animal were examined for their restriction-fragment-length polymorphism, showing that they were classified into four major types. Sequence analysis revealed six types of DNA sequences encoding major piroplasm surface protein with homologies of between 75 and 91%. Of the six sequences, four were identical to those previously reported, while the other two appeared to be new sequences. Among the DNA clones derived from a single infected animal, two to three distinct sequences were often found. Phylogenetic analysis of the six major piroplasm surface protein sequences indicates that five of the six are closely related to each other, and that all are distantly related to the homologous genes of Theileria annulata and Theileria parva. The results suggest that, in addition to those described as T. sergenti/T. buffeli/T. orientalis, there may be some undefined Theileria species distributed in East Asia, and that many cattle are infected with mixed populations of geographically variable Theileria parasites.
The t(10;11)(p13;q14-21) is a non-random translocation that occurs primarily in T cell acute lymphoblastic leukemias (T-ALL), but has also been observed in leukemias and lymphomas of diverse lineages. In U937, a cell line established from a diffuse histiocytic lymphoma, a t(10;11)(p13;q14-21) fuses AF10 to CALM. AF10 is also fused to MLL by a translocation that appears quite similar at the cytogenetic level, the t(10;11)(p12;q23). Fluorescence in situ hybridization studies have demonstrated that AF10 and CALM are also involved in other hematological malignancies containing t(10;11)(p13;q21), but no data are available concerning the molecular details of AF10-CALM fusion in primary leukemias. Using RT-PCR, we amplified multiple different isoforms of AF10-CALM and CALM-AF10 fusion cDNAs from a primary T cell ALL containing a t(10;11)(p13-14;q14-21). These cDNAs arose via alternative splicing of exons from both AF10 and CALM, which we demonstrated can also occur in the native genes. We identified at least two novel AF10 exons that can be included in wild-type and fusion cDNAs. The majority of the AF10 and AF10-CALM cDNA isoforms that we identified are predicted to encode for truncated AF10 polypeptides, raising the possibility that these might have important cellular functions in normal and malignant cells, perhaps by acting as dominant negative inhibitors of full-length AF10 or related proteins.
A chimpanzee (C-499) infected for more than 9 years with two subtype B isolates of human immunodeficiency virus type 1 (HIV-1), one (HIV-1(SF2)) that replicates poorly and one (HIV-1(LAV-1b)) that replicates efficiently in chimpanzees, died of AIDS 11 years after initial infection (F. J. Novembre et al., J. Virol. 71:4086-4091, 1997). Nucleotide sequence and phylogenetic analyses of the C2 to V5 region of env (C2-V5env) in proviral DNA from peripheral blood lymphocytes obtained 22 months before death revealed two distinct virus populations. One of these populations appeared to be a recombinant in env, having the V3 loop from HIV-1(SF2) and the V4-V5 region from HIV-1(LAV-1b); the other population had evolved from HIV-1(LAV-1b). In addition to C2-V5env, the entire p17gag and nef genes were sequenced; however, based on nucleotide sequences and phylogeny, whether the progenitor of the p17gag and nef genes was SF2 or LAV-1b could not be determined. Compared to the two original viruses, the divergence of all clones of C2-V5env ranged from 9.37 to 20.2%, that of p17gag ranged from 3.11 to 9.29%, and that of nef ranged from 4.02 to 7.9%. In contrast, compared to the maximum variation of 20.2% in C2-V5env for C-499, the maximum diversities in C2-V5env in proviruses from two chimpanzees infected with HIV-1(LAV-1b) for 9 and 10 years were 9.65 and 2.48%, respectively. These results demonstrate that (i) two distinct HIV-1 populations can coexist and undergo extensive diversification in chimpanzees with progressive HIV-1-induced disease and (ii) recombination between two subtype B strains occurred even though the second strain was inoculated 15 months after the first one. Furthermore, evaluation of env genes from three chimpanzees infected with the same strain suggests that the magnitude of HIV-1 diversification could be related to higher viral burdens, manifestations of disease, and/or dual infection.
An Egyptian hospital-based pilot case-control study was conducted to investigate the relationship between the expression level of mismatch repair (MMR) genes and the risk of colorectal cancer. The relative expression of five known MMR genes, i.e., hMSH2, hMLH1, hPMS1, hPMS2, and GTBP/hMSH6, was measured by a multiplex reverse transcriptase (RT)-polymerase chain reaction (PCR) in peripheral blood lymphocytes from 31 colorectal cancer patients and 47 age- and-sex matched controls. The expression of hMSH2, GTBP/hMSH6, hPMS1 and hPMS2 tended to be lower in patients than controls, but only the difference in hPMS2 expression was statistically significant (p<0. 01). Although 50% of the cases had chemotherapy or radiotherapy within the last six months before the blood was drawn, their gene expression was not statistically different from those who had not undergone such therapies. After adjustment for age and sex, the odds ratios (OR) calculated from a logistical regression model, using the median levels of gene expression of controls as cut-off values, indicated that increased risk was associated with reduced expressions of both hPMS1 (OR = 3.97, 95% confidence interval (CI) = 1.04 to 7.65) and hPMS2 (OR = 2.86, 95% CI = 1.05 to 7.76). Although the results of this study were inconclusive because of the small sample size and use of prevalent cases, it is biologically plausible that patients with colorectal cancers may have a lower expression of MMR genes than healthy controls because malfunction of these genes has been shown in hereditary nonpolyposis colon cancer. The involvement of low hPMS2 expression in colon cancer risk seems to be unique in the Egyptian population. Further studies with newly diagnosed patients before they begin therapy will provide more convincing data about the role of MMR gene expression in the etiology of colorectal cancers in Egypt.
The incidence of bladder cancer has increased in the United States during the past 50 years, consistent with increased exposure to bladder carcinogens in the environment and tobacco use. Although N-(4-hydroxyphenyl) retinamide (4-HPR), a retinoid derivative, has been used as a chemopreventive agent of bladder cancer in clinical trials, little is known about its mechanisms of action against bladder cancer cells. Previous studies suggest this chemopreventive agent may inhibit tumor growth by inducing apoptosis. To further investigate this putative effect, we examined the effect of 4-HPR and gamma-radiation and their combined effects in three selected bladder cancer cell lines. Indeed, 4-HPR induced apoptosis in these cell lines in a dose-dependent manner. A 2.5 microM dose of 4-HPR and 50 rad of gamma-irradiation induced about 10% increase in apoptotic cells, respectively. However, this low dose 4-HPR combined with low dose gamma-irradiation had a synergistic effect on apoptosis, in which apoptotic cells increased by more than 30%. The findings have potential clinical implications and warrant further investigations both in vitro and in vivo in bladder cancer.
AIM: To overexpress human mu-opioid receptor (muOR) with characteristics similar to those of mammalian origin. METHODS: Human muOR with a tag of 6 consecutive histidines at its carboxyl terminus was expressed in recombinant baculovirus infected Sf9 insect cells. Then the pharmacological characterizations of the product were studied by receptor binding assay and cAMP assay. RESULTS: The maximal binding capacity for the [3H]diprenorphine and [3H]ohmefentanyl (Ohm) were 9.1 +/- 0.7 and 6.52 +/- 0.23 nmol/g protein, respectively. The [3H]diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by alpha-selective agonists [D-Ala2, N-methyl-Phe4, glyol5] enkephalin (DAGO), Ohm, and morphine, but neither by the delta-selective agonist [D-Pen2, D-Pen5] enkephalin (DPDPE) nor by the kappa-selective agonist ¿trans-(+/-)-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]¿ benzacetamide (U50488). NaCl 100 mmol.L-1 and guanosine triphosphate (GTP) 50 mumol.L-1 could reduce mu agonists Ohm and etorphine affinity binding to the expressed muOR. DAGO and Ohm effectively inhibited forskolin-stimulated cAMP accumulation. This agonist-dependent effect was blocked by opioid antagonist naloxone. CONCLUSION: The overexpression of human muOR with a tag of six consecutive histidines at its carboxyl terminus in Sf9 insect cells retained the characteristics of wild-type human muOR.
On the basis of TCM diferentiation, the authors have treated 359 adult female cases of non-obesity with undesirable body shape by combined application of body acupuncture and moxibustion and auricular acupuncture, and achieved quite good remoulding and orthopedic results, suggesting that acupuncture and moxibustion can very effectively regulate the somatotypic indexes of body weight, circumference of the chest, loin, hip and thigh, the ratio of the loin to hip, sebum thickness, obesity degree, body mass index and body fat percentage of the cases in the overweight group and orthopedic groups 1, 2. It is therefore concluded that acupuncture and moxibustion is a good therapy for obtaining a graceful body shape.
Microvessel quantity(MVQ), expression of CD44V6 and EGFR were studied in 48 patients with invasive breast carcinoma by CD31 immunohistochemistry. Significant differences of MVQ, CD44V6 and EGFR were observed (P < 0.01) between the groups with and without metastasis. The results suggest that angiogeneses and activation of tumor metastasis associated gene play an important role in tumor metastasis.
Expression of CD44V6 and nm23-H1 in thyroid papillary adenocarcinoma (TPC) was studied in 84 patients by immunohistochemistry. The positive rates of CD44V6 and nm23-H1 were 63.1% and 48.8% respectively. An increased CD44V6 and a decreased nm23-H1 positive rate in patients with lymph node metastasis (LNM) were observed (P < 0.01). Negative correlation was existed between overexpression of CD44V6 and low-expression of nm23-H1 (P < 0.01). The results suggested that CD44V6 and nm23-H1 play important role in LNM of TPC and disordered expression of CD44V6 and nm23-H1 are involved in the process of LNM.
A new indicator reaction is developed on the basis of manganese catalyzed oxidation of coomassie brilliant blue G250 by potassium periodate with nitrilotriaetic acid as an activator. The detection limit for manganese is 6.76 x 10(-8) g/L. The linear range of determination is 0.02-0.30 microgram/25 mL.
To know well the prostate symptoms in old men, the prostate symptom scores of 412 male residents over 60 years of age in Chengdu area (116 in city and 296 in countryside) were investigated by using I-PSS. The results showed that there was a significant difference between the city and countryside in terms of symptoms score(P < 0.05) and quality of life scores (P < 0.05). No significant difference was noted among the three age groups (P > 0.05). However, the mean score and the percentage of high score section in the elder group were higher than those in the younger group. The prevalence of benign prostate hypertrophy (BPH) and the demands on quality of life in city were different from those in countryside, and the prevalence rate of BPH increased with age.
Head and neck cancer, like lung cancer, is considered a paradigm of an environmentally induced disease. Genetically determined variation in DNA repair capacity is thought to contribute to susceptibility to tobacco-related cancers. In this molecular epidemiology study, we investigated the association between DNA mismatch-repair (MMR) gene expression and the risk of head and neck cancer. Using our newly developed multiplex reverse transcription-PCR assay, we simultaneously evaluated the relative expression levels of five MMR genes (hMSH2, hMLH1, hPMS1, hPMS2, and hGTBP/hMSH6) in the peripheral blood lymphocytes of 78 patients (mean age = 59.6 +/- 12.4 years) with newly diagnosed head and neck cancer and 86 healthy controls (mean age = 58.2 +/- 12.9 years). The relative MMR gene expression was not correlated with disease stage or tumor site in the cases or with smoking and alcohol use in the controls. The expression levels increased with age in both cases and controls, but the mean expression of hMLH1, hPMS1, and hGTBP/hMSH6 was significantly lower in the cases than in the controls (P < 0.05). Using the median expression level in controls as the cutoff value, significantly increased odds ratios (ORs) were associated only with low expression of hMLH1 (OR = 4.4; 95% confidence interval = 2.1-9.1) and hGTBP/hMSH6 (OR = 2.1; 95% confidence interval = 1.1-4.1) after adjustment for age, sex, ethnicity, smoking status, and alcohol use. The results suggest that low hMLH1 and hGTBP/hMSH6 expression is associated with an increased risk of head and neck cancer. Additional studies with a larger number of subjects are warranted to confirm these findings.