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Biomedical subjects

Z Gu

Publications and source records attributed to Z Gu.

At least 199 records · Page 11Linked to original sources

NIP1, a gene required for nuclear transport in yeast.

Cytochrome c with a nuclear localization signal added at the N terminus was mistargeted to the nucleus, resulting in a yeast strain deficient in mitochondrial cytochrome c. Reversion of this strain allowed the isolation of temperature-conditional mutants defective in nuclear transport, as demonstrated with one of these mutants, nip1-1, that was shown to be defective in nuclear accumulation of a LacZ protein containing a nuclear localization signal of the yeast ribosomal protein L29. The NIP1+ gene was cloned and shown to encode a 93,143-Da protein. Furthermore, an epitope-labeled NIP1 protein migrated in SDS/polyacrylamide gels with a mass of approximately 100,000 Da and was shown by immunofluorescence to localize mainly in the cytoplasm. NIP1+ was shown to be an essential gene by gene disruption experiments. Intriguingly, NIP1 has a serine-rich acidic N-terminal region that is similar in this regard to the N-terminal region of a previously described nuclear localization signal-binding protein, NSR1.

Alleles↗

Nucleolin forms a specific complex with a fragment of the viral (minus) strand of minute virus of mice DNA.

Nucleolin, a major nucleolar protein, forms a specific complex with the genome (a single-stranded DNA molecule of minus polarity) of parvovirus MVMp in vitro. By means of South-western blotting experiments, we mapped the binding site to a 222-nucleotide motif within the non-structural transcription unit, referred to as NUBE (nucleolin-binding element). The specificity of the interaction was confirmed by competitive gel retardation assays. DNaseI and nuclease S1 probing showed that NUBE folds into a secondary structure, in agreement with a computer-assisted conformational prediction. The whole NUBE may be necessary for the interaction with nucleolin, as suggested by the failure of NUBE subfragments to bind the protein and by the nuclease footprinting experiments. The present work extends the previously reported ability of nucleolin to form a specific complex with ribosomal RNA, to a defined DNA substrate. Considering the tropism of MVMp DNA replication for host cell nucleoli, these data raise the possibility that nucleolin may contribute to the regulation of the parvoviral life-cycle.

Base Sequence↗

Perturbing highly conserved spatial relationships in the regulatory domain that controls inducible cat translation.

Chloramphenicol activates translation of cat-86 mRNA by stalling a ribosome in the leader of individual transcripts. Stalling triggers two sequential events: the destabilization of a region of secondary structure that sequesters the cat ribosome-binding site (RBS-C), and the initiation of cat translation. The site of drug-dependent ribosome stalling is dictated by the leader sequence, crb; crb causes a ribosome to stall with its aminoacyl site at leader codon 6. We demonstrate that induction requires the maintenance of a precise spatial relationship between crb and sequences within the left inverted repeat of the secondary structure. Therefore, destabilization of the secondary structure during chloramphenicol induction may result from the interaction of a stalled ribosome with a specific sequence in the secondary structure rather than from non-specific masking of RNA sequences. cat-86 regulation also depends on the distance that separates crb from RBS-C. This interval of 33 nucleotides was incrementally increased and decreased by mutations within a loop in the secondary structure. Shortening the distance between crb and RBS-C by three nucleotides reduced induction by half and a deletion of nine nucleotides abolished induction. Insertion mutations were without effect on induced expression but elevated basal expression. The results indicate that when the A site of a ribosome occupies leader codon 6 the secondary structure is destabilized and there is no interference with entry of a second ribosome at RBS-C. The data further demonstrate that when the A site of a ribosome in the leader is within 30 nucleotides of RBS-C, cat expression decreases. This decrease probably results from competition of the leader ribosome with the ribosome initiating cat translation. Our observations demonstrate that in wild-type cat-86 the distances between crb and the secondary structure, and between crb and RBS-C provide the precise spacing necessary to achieve three interdependent effects: the destabilization of the RNA secondary structure by a ribosome stalled at crb; a lack of competition between a ribosome stalled at crb and the initiating ribosome; and maintenance of a low, but measurable, basal level of cat expression. The spatial relationships identified as necessary for the regulation of cat-86 are conserved in the regulatory regions for five other inducible cat genes.

Base Sequence↗

Novel mutation in the human immunodeficiency virus type 1 reverse transcriptase gene that encodes cross-resistance to 2',3'-dideoxyinosine and 2',3'-dideoxycytidine.

We have used the technique of in vitro selection to generate variants of human immunodeficiency virus type 1 (HIV-1) that are resistant to 2',3'-dideoxyinosine (ddI) and cross-resistant to 2',3'-dideoxycytidine (ddC). The complete reverse transcriptase (RT)-coding regions, plus portions of flanking sequences, of viruses possessing a ddI-resistant phenotype were cloned and sequenced by polymerase chain reaction (PCR)-based methods. We observed that several of these viruses possessed mutations at amino acid sites 184 (Met-->Val; ATG-->GTG) and 294 (Pro-->Ser; CCA-->TCA). These mutations were introduced in the pol gene of infectious, cloned HXB2-D DNA by site-directed mutagenesis. Viral replication assays confirmed the importance of site 184 with regard to resistance to ddI. The recombinant viruses thus generated displayed more than fivefold-greater resistance to ddI than parental HXB2-D did. Moreover, more than fivefold-greater resistance to ddC was also documented; however, the recombinant viruses continued to be inhibited by zidovudine (AZT). No resistance to ddI, ddC, or AZT was introduced by inclusion of mutation site 294 in the pol gene of HXB2-D. PCR analysis performed on viral samples obtained from patients receiving long-term ddI therapy confirmed the presence of mutation site 184 in five of seven cases tested. In three of these five positive cases, the wild-type codon was also detected, indicating that mixtures of viral quasispecies were apparently present. Viruses possessing a ddI resistance phenotype were isolated from both subjects whose viruses contained only the mutated rather than wild-type codon at position 184 as well as from a third individual, whose viruses appeared to be mostly of the mutated variety.

Acquired Immunodeficiency Syndrome↗

[Application of Bayes discriminatory analysis to malaria stratification in Hainan province].

The present study was conducted to explore a new method for stratification of malaria endemicity. Several socioeconomic and geographic factors deemed appropriate to be employed in association with malaria epidemic potential were deal with Bayes discriminatory analysis.55 local villages in 12 counties of Hainan province were selected for socioeconomic survey, among which 33 villages were taken as the modelling sample and 22 villages were the non-modelling sample. The questionnaire contained a total of 12 relevant items. After socioeconomic inquiry was performed, the data were analysed through Bayes discriminatory theorem. Six important factors ware selected, i.e. topography (X1), proportion of labour forces (X4), income per person (X5), house structure (X8), knowledge of malaria control (X10), frequency of over-night stay in mountain forest (X12). Three discriminatory function formulas referring respectively to hyper-, meso-, and hypo-endemicity were set up. These 3 discriminatory function formulas were further applied to fit the endemicity of 33 model-sampling villages and 22 non-model-sampling villages. The agreement rate were 91.0% and 77.3% respectively.

Bayes Theorem↗

Generation of nucleoside-resistant variants of HIV-1 by in vitro selection in the presence of AZT or DDI but no by combinations.

Several laboratories have shown that AZT-resistant variants of HIV-1 can be isolated from patients who have received prolonged therapy with this drug. Our laboratory has now been able to generate HIV-1 variants resistant to both AZT and ddI, in tissue culture, by using step-wise increases in the concentrations of each of these compounds over a 10-week period. This work has been performed by culturing wild-type clinical strains of HIV-1 as well as the HIV-3b laboratory strain of this virus under such conditions. The ID50 values obtained for the resistant viruses thus generated vary between 50-100 times above those of the parental wild-type strains in each case. Furthermore, we have identified several new mutation sites in the HIV-1 pol gene that are responsible for the observed resistance to AZT and ddI. We have not succeeded, however, in generating drug-resistant strains of HIV-1, under conditions in which several compounds or anti-viral agents were simultaneously present during the in vitro selection process. Combinations of drugs which failed to yield drug-resistant variants included AZT plus ddI, AZT plus alpha-interferon, and ddI plus alpha-interferon. These findings indicate that HIV drug resistance is less likely to occur in tissue culture when combinations of drugs are used, and provide rationale for the development of combination clinical trials for treatment of HIV-associated disease.

Amino Acid Sequence↗

Diagnostic exploration of enlarged peripheral nerves in suspected cases of leprosy. An analysis of 55 cases.

In 55 cases presenting with enlarged peripheral nerves without any skin lesions, a rice grain-sized biopsy of the nerve lesion was taken for histopathological examination. As a result definitive diagnoses could be established: leprosy was diagnosed in 32 cases. In 23 cases the cause of nerve enlargement was not leprosy: post-traumatic neuritis 9, cysts 5, hypertrophic neuritis 3, nonspecific 4, neurofibroma 1, and amyloidosis 1. In all of these cases there was a deficit of the nerve function and postoperatively there were no complications. The authors, as a result of this experience, believe that surgical exploration and biopsy is a harmless diagnostic tool for establishing a definitive diagnosis of leprosy in cases presenting with enlarged peripheral nerves without any skin lesions. In 23 out of 55 such cases the nerve enlargement was proved to be other causes than leprosy.

Adolescent↗

Observation of the activity of factor VIII in the endometrium of women pre- and post-insertion of three types of IUDs.

The endometrial materials were obtained from 90 women who had been randomly inserted with three types of IUDs (Stainless steel ring, SS; copper T 220, TCu 220, and levonorgestrel-releasing device, LNG). An immunoperoxidase reaction, PAP method, with the antiserum of Factor VIII as the primary antibody, was carried out to detect the Factor VIII activity in the endometrial endothelium pre- (control) and post-insertion of the IUDs. The results revealed that: 1. There was a generalized lower Factor VIII activity in the endometrium of women post-insertion of IUDs (except LNG). 2. Comparison of the Factor VIII activity in the endometrium of women using different types of IUDs showed that the TCu type and the SS type decreased the activity significantly whereas the activity remained unchanged after 3-6 months' use of the LNG-IUD. The different types of IUDs seemed to influence the coagulation regulatory system in different ways; the lower Factor VIII activity, the more tendency to bleeding. 3. The Factor VIII activity in the endometrium of women using IUDs was lower in all phases of the menstrual cycle including the proliferative phase when the Factor VIII activity is normally high. It cannot be excluded that this could be a contributing factor to IUD-induced bleedings.

Adult↗

[Clinical study of superoxide dismutase in recurrent aphthous ulceration].

Activity of superoxide dismutase (SOD) was determined in erythrocytes obtained from 74 patients with recurrent aphthous ulcerations (RAU) and 100 healthy individuals. The results showed that activity level of SOD in RAU was significantly lower as compared with that in the controls (P less than 0.01). The activity of SOD in 30 cases of RAU at interval period was elevated on reexamination, but it was still lower than that in the controls (P less than 0.01). There is evidence that damage of superoxide radicals (O.2) and decrease of activity of SOD may play important roles in the pathogenesis of RAU. A random double-blind, placebo-controlled, cross-over trial on membrane SOD was carried out in 40 patients with RAU. The results demonstrated that the effective rate of SOD was 90%, being markedly higher than that of placebo. It is suggested that membrane SOD, as a topical medicine, could increase the concentration of SOD on the ulcerous surfaces, prevent O.2-induced injury, decrease capillary permeability, relieve inflammation and improve cure of ulcer.

Administration, Topical↗

[Analysing the effects of tooth extraction under acupuncture anesthesia in 825 cases of senior].

It has been found that the effects of tooth extraction under acupuncture anesthesia (AA) for the old patients are generally quite good. In order to verify this result, the authors had statistically analysed 825 cases aged over 60 years (Group A) and equal quantity whose age were between 18-40 years old (Group B) out of more than 4000 cases whose records of AA were rather completely during 1973 to 1988. 1. There was no significant difference under AA for different sexes. 2. The differences of the excellent rate of AA between Group A and Group B were extremely evident either in the positions of the tooth or the reason of tooth extraction. The former was 89.04% and the latter 76.20% (P less than 0.005). 3. Although a low rate in excellence of tooth extraction under simple AA (only 75.58%), not so good as the effect by AAA (an average about 88.21%), the simple AA was more common and more easier to be accepted by patients. 4. Tooth extraction under AA has been proved to be safe, effective and without any complications. 5. The reasons for the senior to extract their teeth were almost for the broken crowns and of roots or the purposes of prosthetic restoration. Even though the pain thresholds in the senior were commonly higher. It is still an important factor of the good effect and good rate of tooth extraction under AA. Referring to the statistics in this study the authors suggest that the hospitals which can use the AA for tooth extraction adopt the method as the first choice of anesthesia method to the senior who intends to accept it.

Acupuncture Analgesia↗

Observation of the activity of factor VIII in the endometrium of women with regular menstrual cycles.

An immunoperoxidase reaction was used to detect Factor VIII activity in the endometrial endothelium during the normal menstrual cycle. Fifty-three women of fertile age, having regular menstrual cycles, participated in the study. Endometrial material was obtained from 33 women in the preovulatory phase and from 20 women in the post-ovulatory phase. The immunoperoxidase reaction was evaluated using a scoring system. Cyclic changes related to the pre- and post-ovulatory phase were observed in the Factor VIII activity of the endometrial endothelial cells; a gradual increase occurred from the early to the late proliferative phase and thus activity gradually decreased during the secretory phase. No activity was observed in the late secretory or in the menstrual phase.

Adult↗