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Biomedical subjects

L Hu

Publications and source records attributed to L Hu.

At least 145 records · Page 8Linked to original sources
PubMed1996

[The PCR amplification, cloning, sequencing, expression in E. coli of gene encoding endoflagella subunit protein (fla B) from Leptospira interrogans serovar lai].

A pair of oligonucleotide primers were designed by ourselves to amplify the endoflagella gene of L. interrogans serovar lai. A fragment about 840 bp was generated with PCR and inserted into plasmid pUC8 after the fragment and pUC8 were digested respectively with Bam HI and Pst I. A recombinant plasmid (designated as pLF1) was obtained. SDS-PAGE analysis indicated that a 33 kd was expressed in E. coli JM103 harboring pLF1 and the expression level of the protein was 11% of total bacterial soluble proteins. Western blot analysis showed that the protein band could be recognized by the antiserum against the endoflagella (Axiall filament) of Leptospira interrogans serovar lai. Nucleotide seguence data showed an open reading frame encoding 282 aminoacids residues, corresponding to a protein of molecular weight 33.6 kd. The G + C content of endoflagella subunit protein gene was 48 mol%. Therefore, the G + C content of the leptospiral fla B Gene is significantly higher than the reported 39 mol% G + C content of leptospiral genome of L.interrogans serovar lai but similar to the G + C of the Treponema pallidum genome. Comparison of the deduced endoflagellar subunit protein (fla B) amino acid sequence with flagellins from other bacteria revealed a high level of identity with the Treponema pallidum fla B proteins. Immunization/protection experiment was performed on the model of BALB/c mice and showed that the survival rate in the group JM103-pLF1 was higher than that in the group JM103-pUC8, but statistically the difference between them was significant (P < 0.05) and pLF1 did not induce significant levels of agglutinating antibodies against L.interrogans serovar lai.

Amino Acid Sequence↗
PubMed1996

Studies on prophylactic effect of artesunate on schistosomiasis japonica.

OBJECTIVE: To examine the prophylactic effect of artesunate on schistosomiasis japonica by killing schistosomula. METHODS: Mice, rabbits and dogs after infection with cercariae of schistosoma japonicum (S. japonicum) were treated with artesunate on the 7th day at a dose of 300 mg/kg, 20-40 mg/kg and 30 mg/kg respectively once a week for 4-6 weeks. A double-blind test was used. A total of 864 persons in highly endemic areas for schistosomiasis were administered either artesunate at a dose of 6 mg/kg once a week for 8 weeks or identical placebo with the same dose-schedule during transmission season for S. japonicum. Four weeks after the last dosing, fecal examinations for S. japonicum eggs and miracidia were carried out to evaluate the prophylactic effect. RESULTS: Worm reduction rates in mice, rabbits and dogs were 77.50-90.66%, 99.53% and 97.10% respectively. All of the 467 residents in 2 trials were free from eggs or miracidia upon stool examinations whereas in the control groups with placebo, 15 out of 218 (6.9%) and 26 out of 179 (14.5%) were stool eggs and/or miracidia positive in the first and second trial, respectively. Side effects were mild. No significant changes in routine blood and urine tests, electrocardiogram, hepatic and renal functions were observed after artesunate administration. CONCLUSIONS: Animal experiments and field trials have demonstrated good efficacy of artesunate on killing schistosomula with little side effects. Thus, the drug is suggested to be used on the population in endemic areas for prevention of schistosomiasis.

Adolescent↗
PubMed1996

[Experimental study of the protective effects of astragalus and salvia miltiorrhiza bunge on glycerol induced acute renal failure in rabbits].

The astragalus and salvia miltiorrhiza bunge alcohol extracts were used in preventive treatment of glycerol induced acute renal failure (ARF) in rabbits. The experimental rabbits were divided: astragalus group (AB), salvia miltiorrhia bunge group (SM), two extracts mixture group (ABSM), and normol saline control group (C). The extracts, mixtures and normol saline were administrated before and after the induced ARF. The counts of Na, K, creatinine in urine and blood and urinary AAP, NAG were determinated during 24-14 days periodically in all 4 groups, and the renal tissues were taken from same periods for pathomorphological studies by microscopy and transmission electromicroscopy. The damage of the glycerol induced ARF was not only in the convolute tubules but also in the glomerulus. The glomerulor filtration rate reduced abruptly, and oligouria or auria developed. The study of renal functions and renal morphogy showed that the AB and ABSM groups were damaged more lesser and promptely repaired than the SM and C groups in the early stage. No glomerular sclerosis was noted in the AB, ABSM groups in later stage, but it occurred sporadically or diffusely in the SM, C groups. Therefore, astragalus is an ideal protective drug of traditional chinese medicine for ARF.

Acute Kidney Injury↗
PubMed1996

[Research on the recombinant plasmid pDJH2 of L. interrogans serovar lai: sequencing and alignment with other known bacterial Omp sequence].

The Leptospira whole cell vaccine (LWCV) currently used in China is safe and effective, out the immunity following vaccination with two doses of the fluid medium vaccine is of low order. The duration of immunity conferred by this vaccine is rather short, six months or at most one year. Therefore, it is necessary to develop new generation vaccines against Leptospirosis for the developing world. In this paper we report the sequencing of the insert fragment of pDJH2 from genomic DNA of L. interrogans sevovar lai strain 017 and its alignment with other bacterial omp sequences. A genomic library of Leptospira interrogaans serovar lai strain 017 was constructed with the plasmid vector pUC18. A recombinant plasmid designated pJDH2 was screened from the genomic library. Inserted fragment of pDH2 is 1.9 kb by gel electrophoresis. Immunization/protection was studied in BALB/c mice model. The results showed highly significant difference between pDJH2 and pUC18 (control). Inserted fragment of pDJH2 DNA sequencing was performed by Dr Yan Zhengxin (Max-Planck-Institut for Biology. Tubingen, Germany). Insert fragment was cloned into pBluescript II KS-(stratagene) and sequenced by using AB1 (Applied Bio Systems, Model 373A). Two open reading frames of 565 and 662 nucleotides were identified. There were identifiable initiation codons, terminators, Shine-Dalgano ribosome combining site, Pribnow boxes and Sextama boxes within the 2 sequenced regions. Nucleotide sequences were analysed using Gene Work, a suit of computer program developed by Department of Biochemistry St. Jude Children's Research Hospital Memphis. U.S.A. The results of formatted alignment showed the predicted nucleotide sequence of ORF1 of the serovar lai had significant similarity with ORF2 (49.36%). L. kirschneri ompL1 (49.26%), Borrelia burgdoferi omp (48.97%), Treponema phagedenis omp (47.3%); Salmonella typhimurium ompC(46.87%), Yersinia enterocolitica ompH (46.7%), Leptospira borgpeterseni pfap (46.3%), and Serratia marcescens omp (43.3%). The close relationship of the pDJH2 ORF1 and ORF2 nucleotide sequences from Leptospira kirschneri ompL 1 is apparent. Whether the recombinant pDJH2 will prove useful for vaccine development remains to be tested.

Animals↗
PubMed1996

[Subclones of fragment DNA recombinant rpDJH2 of L. interrogans serovar lai strain 017 and it's expression of high level in E. coli].

Fragment of 1.9 kb recombinant DNA of pDJH2 was linked with vectors pT7-7 and pRSETs. Then they were transformed into E. coli JM109 (DE3) respectively. Expression of subclones was achieved in E. coli JM109 (DE3) with IPTG inducement. SDS-PAGE showed that the molecular weights of products were 68kd and 23 kd respectively. The amount of production seemed to be higher than that of the outer membrane proteins of L. interrogans serovar strain 017 in nature. Immunoblotting of pDJt and pDJrB2 (both are subclones) with the specific antiserum of anti-OMP of L. interrogans serovar lai strain 017 and the experiment of initiative immuno-protection in guinea pigs showed both protein-68 kd and 23 kd might be the antigens of immuno-protection on the outer membrane of L. interrogans serovar lai strain 017.

Animals↗
PubMed1996

[The role nitric oxide and other neurotransmitter in canine penile erection].

The role on nitric oxide and its relative factors (cGMP, cAMP, methylene blue) was studied in canine erection induced by stimulating pelvic nerves, and the effect of cholinergic neuroeffectors and the sinusoidal endothelium was also observed in this experiment. The results indicate that intracavernous injection of nitric oxide can evoke a penile tumescence, which is similar to that of the neurostimulation. The results also suggest that the cholinergic nerves and the sinusoidal endothelium are involved in erection, and the effect of the former must depend on mediation of the latter. The study supports that cholinergic and nonadrenergic noncholinergic (NANC) neuroeffectors take part in penile erection, and nitric oxide may be one of chief NANC neurotransmitters.

Animals↗
PubMed1995

Monobromobimane as an affinity label of the xenobiotic binding site of rat glutathione S-transferase 3-3.

Monobromobimane (mBBr), besides being a substrate in the presence of glutathione, inactivates rat liver glutathione S-transferase 3-3 at pH 7.5 and 25 degrees C as assayed using 1-chloro-2,4-dinitrobenzene (CDNB). The rate of inactivation is enhanced about 5-fold by S-methylglutathione. Substrate analogs bromosulfophthalein and 2,4-dinitrophenol decrease the rate of inactivation at least 20-fold. Upon incubation for 60 min with 0.25 mM mBBr and S-methylglutathione, the enzyme loses 91% of its activity toward CDNB and incorporates 2.14 mol of reagent/mol of subunit, whereas incubation under the same conditions but with added protectant 2,4-dinitrophenol yields an enzyme that is catalytically active and contains only 0.89 mol of reagent/mol of subunit. mBBR-modified enzyme is fluorescent, and fluorescence energy transfer occurs between intrinsic tryptophan and covalently bound bimane in modified enzyme. Both Tyr115 and Cys114 are modified, but Tyr115 is the initial reaction target and its modification correlates with loss of activity toward CDNB. The fact that the activity toward mBBr is retained by the enzyme after modification suggests that rat isozyme 3-3 has two binding sites for mBBr.

Affinity Labels↗
PubMed1995

Radiation-induced changes in nucleoid halo diameters of aerobic and hypoxic SF-126 human brain tumor cells.

Nucleoid halo diameters were measured to assay changes in DNA supercoiling in human brain tumor cell line SF-126 after irradiation under aerobic or hypoxic conditions. In unirradiated aerobic cells, a typical propidium iodide titration curve showed that with increasing concentrations of propidium iodide, the halo diameter increased and then decreased with the unwinding and subsequent rewinding of DNA supercoils. In irradiated cells, the rewinding of DNA supercoils was inhibited, resulting in an increased halo diameter, in a radiation dose-dependent manner. To produce equal increases in halo diameter required about a threefold higher radiation dose in hypoxic cells than in aerobic cells. Quantitatively similar differences in the radiation sensitivities of hypoxic and aerobic cells were demonstrated by a colony-forming efficiency assay. These findings suggest that the nucleoid halo assay may be used as a rapid measure of the inherent radiation sensitivity of human tumors.

Aerobiosis↗
PubMed1995

p53 controls proliferation of early B lineage cells by a p21 (WAF1/Cip1)-independent pathway.

We examined a role of p53 in early B cell development using fetal liver (FL) cells from p53 deficient (-/-) mice. Expression of p53 was detected in early B lineage (pre-B) cells developed in the stromal cell-dependent FL cell culture. Proliferation of the pre-B cells from the p53(-/-) mice was augmented, suggesting that p53 negatively regulates proliferation of pre-B cells. Although p21 (WAF1/Cip1) is a downstream effector of p53 and mediates growth arrest by inhibiting the action of cyclin-dependent kinases, expression of p21 was also induced in the pre-B cells from p53(-/-) mice. These results indicate that expression of the p21 in pre-B cells is independent of p53 and that the regulatory effect of p53 on proliferation of pre-B cells is mediated by a p21-independent pathway.

Animals↗
PubMed1995

Typing of Staphylococcus aureus colonising human nasal carriers by pulsed-field gel electrophoresis.

Colonisation by Staphylococcus aureus in the nares of 120 outpatients and 63 healthy adults was studied for c. 2 years. Two states of carriage of S. aureus were confirmed: persistent carriage and persistent non-carriage. The states of carriage and non-carriage were quite stable and > 60% of the population of any of the study groups were stable non-carriers. The results of typing the strains isolated from the same individuals at different times with DNA fingerprinting by digestion with SmaI enzyme showed that all the stable carriers were persistently infected with the same strain and that changes in the strain seldom occurred.

Adult↗
PubMed1995

Typing of Staphylococcus epidermidis colonizing in human nares by pulsed-field gel electrophoresis.

From the nares of 11 healthy adults, 253 strains of coagulase negative staphylococcus were isolated and 88% of them were identified as Staphylococcus epidermidis using the API STAPH system. Chromosomal DNA fingerprinting of the isolated strains revealed that each person carried multiple types of S. epidermidis in his or her nares. The colonization of the strains was not stable; the types of the isolates changed in the first and the second examinations 5 months apart. The results contrasted with previous findings in which only one strain of S. aureus colonized persistently in the nares of healthy adults.

Adult↗
PubMed1995

Role of nitric oxide in regulation of long-term pressure-natriuresis relationship in Dahl rats.

The aim of this study was to determine the role of nitric oxide (NO) in the development of salt-induced hypertension in the Brookhaven strain of Dahl rats. Six- to seven-week-old conscious salt-sensitive (S) and salt-resistant (R) rats with indwelling arterial and venous catheters received low-, normal-, and high-sodium intakes sequentially over a 16-day period, and L-arginine was infused intravenously at 2 or 4 mg.kg-1.min-1 over this time. The S rats had an impaired NO production as evidenced by a decreased urinary nitrate plus nitrite excretion. The administration of the low or high dose of L-arginine increased the whole body NO production of the S rats to that of the control R rats, and the high dose of L-arginine prevented the shift of long-term pressure-natriuresis relationship, the elevation of arterial pressure, and the increase in salt sensitivity of arterial pressure in the S rats. The sodium and water balances were not different between the age-matched R and S rats. In conclusion, a continuous infusion of L-arginine prevented both the changes in the pressure-natriuresis relationship and the development of salt-induced hypertension in Dahl S rats.

Animals↗
PubMed1995

[An epidemiological study on the characteristics of smoking behaviors among 910 young workers in Shangqiu City, Henan Province].

This paper dealt with an epidemiological investigation on the smoking behaviors of the young people in Shangqiu city, Henan province. The period of their working history was less than 5 years and their ages younger than 30 years. The interviewees were chosen from the hospitals, middle or professional schools, and large factories in the city. There were 587 males and 339 females included in this sample. Everyone was asked to complete the self-administered questionnaire in the workplace. The total number of sample was 926. The rate of respondence was 98.3%. Of the 926 questionnaires, 910 (98.3%) were identified as qualified. The investigation showed that the overall smoking rate was 32.1%, with male 64.2% and female 1.5%, respectively. The smoking rate of male was approaching the highest rate of adult males in our country, much higher than that of adolescents in school. The rates were parallelly increasing with years of working. About 50% of the young smokers began to be habitual since started their job, especially from the first four years of work. More than 80 percent of the smoking young workers were mild or light smokers, and did not smoke in "Ban Smoking Place". However, the most popular places for smoking were the place where there is a sign at their workplaces. They smoke most often with their colleagues and friends.

Adult↗
PubMed1994

Neuropeptide Y N-terminal deletion fragments: correlation between solution structure and receptor binding activity at Y1 receptors in rat brain cortex.

N alpha-Acetyl (Ac), N-terminal deletion fragments of porcine neuropeptide Y (NPY) have been synthesized and characterized for solution conformation properties by circular dichroism and for receptor binding activity at benextramine-sensitive Y1 binding sites in rat brain cortex. Sequential deletion of Tyr1, Pro2, and Ser3 had no effect on the structural (alpha-helical content of 32.5, 30.6, and 30.7%, respectively, at 1 x 10(-5) M) or aggregation (monomer to dimer transition for N alpha-Ac-NPY3-36 and N alpha-Ac-NPY4-36) properties of NPY. In contrast, deletion of Tyr1 decreased receptor binding activity in rat brain cortex by 4-fold (IC50 = 13.0 nM versus 3.75 nM for NPY), but further deletion of Pro2-Ser3 had no additional detrimental effect on receptor binding activity relative to the desTyr1 analog. Thus, Pro2 and Ser3 do not contribute either to the stability of the NPY tertiary structure nor directly to the receptor-ligand interactions. Additional removal of N-terminal amino acids Lys4-Pro5 decreased the helical content and abolished aggregation to a dimeric form of the resultant analog, results suggesting that the residues around Pro5 are important for formation of NPY's compact, pancreatic polypeptide (PP)-fold structure. This loss in structure also correlated with a further 2-3 fold drop in receptor binding activity. These structure-activity correlations provide evidence for the importance of the PP-fold structure in the activity of NPY at Y1 receptors in rat brain cortex.

Amino Acid Sequence↗
PubMed1994

Effect of beta-lymphocyte- and NPC-derived EBV-LMP1 gene expression on in vitro growth and differentiation of human epithelial cells.

The effect of expression of the Epstein-Barr-virus (EBV) latent membrane protein (LMP1) derived from B-lymphocytes (B) and nasopharyngeal carcinoma (NPC) (C) on the in vitro growth and differentiation of a human keratinocyte line, Rhek-1, was analyzed in clonal growth and in in vitro differentiation assays. In contrast to the polygonal parental cells, the B-LMP1-expressing sublines were spindle-shaped while the C-LMP1-expressing cells were pleomorphic. Both B- and C-LMP1-expressing sublines showed increased proliferation as evidenced by: (1) higher colony-forming efficiency (CFE) and larger colony size at reduced serum levels; (2) an increased number of epithelial cell layers formed in the air-liquid-interface culture system and (3) increased expression of proliferative cell nuclear antigen (PCNA). At low serum concentration, the C-LMP1-expressing sublines formed larger colonies than those expressing B-LMP1. In the air-liquid-interface culture system, both B- and C-LMP1-expressing lines showed reduced epithelial differentiation resulting in reduced stratification and reduced involucrin expression similar to those of the cancer cell line, Siha. The results of the present study indicate that the expression of LMP1 in human keratinocytes is associated with morphological transformation and predisposes these cells to a more neoplastic phenotype. The structural difference between the 2 genes responsible for the functional differences and transforming ability will be pinpointed in further experiments.

Antigens, Viral↗