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Biomedical subjects

K Y Zhou

Publications and source records attributed to K Y Zhou.

18 recordsLinked to original sources
PubMed2004

Dynamic depth-dependent osmotic swelling and solute diffusion in articular cartilage monitored using real-time ultrasound.

The objective of this study was to investigate the feasibility of ultrasonic monitoring for the transient depth-dependent osmotic swelling and solute diffusion in normal and degenerated articular cartilage (artC) tissues. Full-thickness artC specimens were collected from fresh bovine patellae. The artC specimens were continuously monitored using a focused beam of 50 MHz ultrasound (US) during sequential changes of the bathing solution from 0.15 mol/L to 2 mol/L saline, 0.15 mol/L saline, 1 mg/mL trypsin solution, 0.15 mol/L saline, 2 mol/L saline and back to 0.15 mol/L saline. The transient displacements of US echoes from the artC tissues at different depths were used to represent the tissue deformation and the NaCl diffusion. The trypsin solution was used selectively to digest the proteoglycans in artC. It was demonstrated that high-frequency US was feasible for monitoring the transient osmotic swelling, solute transport and progressive degeneration of artC in real-time. Preliminary results showed that the normal bovine patellar artC shrank during the first several minutes and then recovered to its original state in approximately 1 h when the solution was changed from 0.15 mol/L to 2 mol/L saline. Degenerated artC showed neither shrinkage nor recovery during the same process. In addition, a dehydrated-hydrated artC specimen showed much stronger shrinkage and it resumed the original state when the solution was changed from 2 mol/L back to 0.15 mol/L saline. The diffusion of NaCl and the digestion process of proteoglycans induced by trypsin were also successfully monitored in real-time.

Animals↗
PubMed2002

Studies on the molecular markers of rhizomes of some Alpinia species.

The rhizomes of Alpinia jianganfeng are used as a traditional Chinese medicine, Jian Gan Feng, to cure rheumatism in Guangdong, China. The rhizomes of some other species of the genus Alpinia such as A. japonica, A. suishaensis and A. nanchuanensis are also used as Jian Gan Feng in Southwest China. However, the identification of the original plants of the crude drugs is difficult. The internal transcribed spacers and the 5.8S coding region of nuclear ribosomal DNA of the four species were sequenced and analyzed. The DNA markers have been determined and they can be used for the molecular identification of these medicinal plants.

Base Sequence↗
PubMed2002

Expression of a novel reticulon-like gene in human testis.

Identification of genes that are specifically expressed in the adult testis or the fetal testis is important for the study of genes related to the development of the testis. In this study, a human testis cDNA microarray was established. PCR products of 9216 clones from a human testis cDNA library were dotted on a nylon membrane; mRNA from adult and fetal testes were purified and probes were prepared by a reverse transcription reaction with testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes, and 96.8 and 95.4% of clones were positive, respectively. In total, 731 clones were differentially expressed: 592 were highly expressed in adult testis and 139 were highly expressed in fetal testis. Among these genes, a new reticulon (Rtn)-like gene was detected and named Rtn-T. Rtn-T was highly expressed in adult human testis. The cDNA of Rtn-T contains 3491 bp and the putative protein had 968 amino acids. This protein is homologous to the six known members of the Rtn family (KIAA0886, Rtn xL, reticulon 4a, Nogo-A, Nogo-A short form, and brain my043) but was different at the 5' end. All homologues originate from one gene, and result from both different promotor regions and different splicing. Rtn-T lacks the first exon and contains a second exon that is lacking in the other homologues. Rtn-T is shorter than KIAA0886, Rtn xL, reticulon 4a and Nogo-A, but longer than the Nogo-A short form and brain my043. Sequence analysis showed that Rtn-T protein has two hydrophobic regions that may be membrane-spanning domains. Expression profiles showed that Rtn-T is specifically and strongly expressed in testis. The results of the present study indicate that the Rtn-T gene is differentially expressed in adult and fetal testes and encodes a membrane protein that may have a function in testis development.

Adult↗
PubMed2001

Characters of nrDNA ITS region sequences of fruits of Alpinia galanga and their adulterants.

The fruits of Alpinia galanga (L.) Sw. are used as a traditional Chinese medicine; but the dry fruits of A. conchigera, A. suishaensis, A. maclurei and A. polyantha are also used as the medicine in local areas. Because dry fruits of these related plants are similar to those of Alpinia galanga (L.) Sw. in odor, morphological characters and chemical components, and even anatomical characters, it is difficult to identify the medicine. Nuclear ribosomal DNA internal transcribed spacer (ITS) regions of the five taxa were directly sequenced using an automated sequencer. Sequence analysis showed that the ITS 1 ranges from 177 to 178 base pairs (bp), and the ITS 2 from 225 to 234 bp. The size of the 5.8S coding region is 164 bp for all species. Also, the pairwise sequence divergence is higher and some molecular markers were determined. According to these molecular markers, Alpinia galanga (L.) Sw. and the related species can easily be distinguished from each other. Therefore, evidence from nrDNA ITS sequence variation can identify the medicine at the DNA level.

Base Sequence↗
PubMed2001

[Nuclear ribosomal RNA small subunit (18S rRNA) nucleotide sequencing and characterization of sailonggu(whole bone of Myospalax baileyi Thomas)].

OBJECTIVE: Sequencing the nuclear ribosomal RNA small subunit (18S rRNA) gene of Myospalax baileyi (Cricetidae) to develop an ultimate and definitive means for origin identification of genuine Sailonggu. METHODS: The total DNA was prepared from dried tail tissues. The nuclear 18S rRNA gene region was amplified by PCR using a consensus primer set and its nucleotide sequence was determined by PCR direct sequencing. The characteristic analysis of 18S rRNA sequences was generated using software program Genetyx-SV/R Version 10.1. RESULTS: The entire 18S rRNA gene region of M. baileyi spanded 1851 bp in length. Although multiple alignment of sequence indicates that there are only lower homology (72.04%-72.18%) comparing with its two alias Mus musculus (GenBank Accession number X00686) and Rattus norvegicus (M11188) (Muridae), their highly conservative domain is located in 1020-1509 nt. There are many variable sites from upstream of 5'-end, which could provide a novel information for molecular recognition of Sailonggu. CONCLUSION: DNA sequencing could be a useful and reliable tool in the origin identification of genuine Sailonggu.

Animals↗
PubMed2001

[Study on allele-specific diagnostic PCR of the traditional Chinese medicines of the deers].

AIM: To develop a convenient and accurate method of DNA molecular marker for the identification of traditional Chinese medicines made of deers, consisting of pilose antler, penis and testis, tendon and foetus. METHODS: Based on the analysis of DNA sequence of mitochondrial Cyt b gene from original animals of both genuine crude drugs, Cervus nippon and Cervus elaphus, and adulterants, a pair of allele-specific primers named as ILu01-L and ILu01-H were designed for distinguishing geniune crude drugs of deers from their adulterants. RESULTS: The results of diagnostic PCR annealing at 64 degrees C for original animals showed that a 365 bp fragment was only amplified from DNA templates of Cervus nippon and Cervus elaphus. For the identification of medicinal materials total of 43 samples from 6 packages were tested under the same reaction conditions except for DNA templates extracted from these crude drugs. Only 9 samples mentioned above was shown to generate positive amplificon. The result indicate that of 8 samples from 1 package of pilose antler and only 1 sample of deer tendon was genuine crude drug. After that, 3 amplified fragments selected randomly were performed with sequencing analysis with the purpose of verifying the results from diagnostic PCR. Data from sequencing confirmed the reliability of diagnostic PCR identification. CONCLUSION: The diagnostic primers designed in the present study were highly specific for Cervus nippon and Cervus elaphus, and they could be used for the authentication of traditional Chinese medicines made from the deer. The quality of the crude drugs of the deer in the current market is a problem and more effective quality control for these traditional Chinese medicines is urgently needed.

Alleles↗
PubMed2001

[rDNA its sequencing of Herba Dendrobii (Huangcao)].

AIM: To study the genetic diversity of ITS sequences of Herba Dendrobii (Huangcao) and analyze the utility of ITS sequences in molecular authentication of Herba Dendrobii (Huangcao) and phylogenetic of Dendrobium. METHODS: The ITS gene fragment was amplified using a pair of primers. The PCR products were purified and sequenced by the methods of Sanger Dideoxy. RESULTS: The DNA sequence of 228-233 bp ITS 1, 242-247 bp ITS 2 gene fragments and 5.8S rDNA were obtained from 14 samples of Dendrobium. The interspecific substitution varies from 11.79% to 31.58% at ITS 1 and 10.29% to 25.30% at ITS 2. The intraspecific substitution of D. nobile is 0.87% at ITS 1 and without difference at ITS 2. The substitution between Dendrobium and outgroup varies from 23.56% to 36.89% at ITS 1 and 26.52% to 33.31% at ITS 2. The phylogenetic tree based on ITS 1 and ITS 2 data was set up. CONCLUSION: The ITS 1 and ITS 2 gene fragments were highly conservative at intraspecific level in Dendrebium, while they were less conservative at interspecific level in D. nobile. They were least conservative between Dendrobium and outgroup. Hence, the sequence of this fragment is a good molecular marker for authentication of the Huangcao. But, further study is necessary for the phylogenetic of Dendrobium.

Base Sequence↗
PubMed2001

[Identification of Chinese crude drug snake gallbladder by DNA molecular marker].

AIM: It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS: DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS: Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION: The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market.

Animals↗
PubMed2000

Isolation and characteristics of autoreactive T cells specific to aggrecan G1 domain from rheumatoid arthritis patients.

Our previous work showed that the cartilage proteoglycan aggrecan could induce an erosive polyarthritis and spondylitis in BALB/c mice and the G1 globular domain of the aggrecan (G1) contained the arthritogenic region. To elucidate whether autoreactive T cells to G1 are expressed in rheumatoid arthritis patients, we analyzed the frequency of human G1-specific T cells in the peripheral blood of five rheumatoid arthritis patients and tried to establish G1-reactive T cell lines from these rheumatoid arthritis patients. The results showed that the G1-specific T cells in PBL were detectable at the range of 4.97 +/- 0.5 x 10(-6) in peripheral blood lymphocytes. We have also generated 15 G1-specific T lymphocyte lines from these patients with a standard split-well method. All these cells expressed fine specificity to human recombinant G1, but not to unrelated antigen. All the 15 lines expressed a pan-T cell marker and 13 of them selectively used the alphabeta T cell receptor. Two of them used gammadelta T cell receptor. The 13 of these T cell lines was CD4 positive. One line expressed CD8. One line expressed both CD4 and CD8. Moreover, 14 out of 15 lines expressed the Th-1 cytokine profile, characterized by interferon-gamma positivity and IL-4 negativity. No Th-2 type cell line was generated. These data provide strong evidence in favor of the presence of autoreactive T cells in the rheumatoid arthritis patients. What is the mechanism(s) that these autoreactive T cells attack self-target and whether these G1-specific, Th-1 type T cell lines can induce arthritis in immune deficiency mice are currently under investigation.

Adult↗
PubMed1999

[RAPD analysis of genetic diversity and phylogenetic relationship of the Taihu pigs].

The genetic diversity and phylogenetic relationship of the Taihu pigs were studied using RAPD technique. A total of 57 individuals were used, and the phylogenetic tree was reconstructed using UPGMA method based on 198 RAPD markers which were amplified with 13 decamer primers. The results indicate as the following: (1) The similarity indexes within each strain and between the strains of the Taihu pigs are all high. The average genetic distance between the strains is 0.052. These results indicate that the Taihu pigs exhibit relatively low level of genetic variation and poor genetic diversity; (2) The Hongdenglong pig is also a strain of the Taihu pigs; (3) The Taihu pigs have close phylogenetic relationship with the Chinese wild boar occurred in the Taihu Lake area; (4) Two lineages diverged in the early stage during the course of the formation of the Taihu pigs. One of the lineage gave rise to the strains of Mizhu, Erhualian, Jiaxinhei and Hongdenglong, another lineage gave rise to the strains of Shawutou, Meishan, Fengjing and Hengjing. The strategy for genetic diversity conservation of the Taihu pigs was discussed.

Animals↗
PubMed1998

Determination of hepatotoxic pyrrolizidine alkaloids by on-line high performance liquid chromatography mass spectrometry with an electrospray interface.

The present study describes the determination of two different types of hepatotoxic pyrrolizidine alkaloids (PAs) and also distinguishing the hepatotoxic PAs from non-toxic ones by both in-source collision-induced dissociation high performance liquid chromatography mass spectrometry (CID-HPLC/MS) and HPLC/MS/MS (CID in the collision cell), using electrospray ionization. The mass spectra provided molecular ions and characteristic fragment ions, which could be used readily for a rapid identification of different types of PAs. Applications of both in-source CID-HPLC/MS and HPLC/MS/MS analytical methods were successful for the determination of PAs in blood samples obtained from rats dosed with PAs and in the PA-containing plant. The results demonstrated that the developed HPLC/MS methods with two different CID techniques provided a very simple and rapid analysis for an unequivocal diagnosis of PA poisoning and for definitive identification of PAs in plants or herbal medicines.

Animals↗
PubMed1998

Application of capillary electrophoresis, liquid chromatography, electrospray-mass spectrometry and matrix-assisted laser desorption/ionization - time of flight - mass spectrometry to the characterization of recombinant human erythropoietin.

High performance capillary electrophoresis (HPCE), high performance liquid chromatography (HPLC), matrix-assisted laser desorption/ionization - time of flight - mass spectrometry (MALDI-TOF-MS), on-line CE-electrospray ionization-mass spectrometry (CE-ESI-MS) and on-line LC-ESI-MS have been employed to characterize a heterogeneous glycoprotein, recombinant human erythropoietin (rHuEPO) expressed from Chinese hamster ovary (CHO) cells. The analysis was demonstrated through two specific levels of detail: the intact protein and tryptic digests of the protein. Six glycoforms of rHuEPO were separated by HPCE; seventeen tryptic fragments in a total of 21 nonglycosylated and glycosylated peptides were characterized; the O-linked glycopeptides were analyzed directly by CE-ESI-MS and LC-ESI-MS. In particular, four glycans of O-acetylation of sialic acid were identified in the O-linked glycosylated fragments. The molecular weight of rHuEPO was accurately determined by MALDI-TOF-MS.

Amino Acid Sequence↗
PubMed1997

[A preliminary study on the identification of crude snake drugs by molecular genetic markers].

DNA was amplified from 11 samples of crude snake drugs by 2 random primers. "Wushaoshe" (Zaocys dhumnades) and its substitutes, Elaphe taeniurus, E. rufodorsata, E. carinata and Sinonatrix annularis, and "Jinqian Baihyashe" (Bungarus multicinctus) and its adulterants, Dinodon rufozonatum and Sinonatrix annularis, can be distinguished by electrophoresis pattern of the amplified products. The results showed that the molecular genetic marker technique can be used as a new method for the identification of crude snake drugs.

Animals↗
PubMed1996

[Isolation and amplification of DNA from tortoise and turtle shells].

DNA was extracted from tortoise and turtle shells, two Chiense crude drugs which have been preserved for at least nine years. The cytochrome b sequence was amplified from the extracts using the polymerase chain reaction. Treatments with detergent and ultraviolet light were employed to destroy possible extraneous contamination. A control extract was set in parallel with the extracts of the specimens in order to detect contamination in solutions and reagents. The results show that significant amounts of genetic information can survive in the shells and may have important implications for their identification.

Animals↗
PubMed1980

Observations on Baiji (Lipotes vexillifer) and finless porpoise (Neophocaena asiaeorientalis) in the lower reaches of the Chang Jiang.

Lipotes usually occurs in pairs or in small loose groups. The calves are born in March-April. The Finless porpoises are usually seen in groups of five or six individuals. The blow of Lipotes lasts 0.2-0.6 sec and that of Neophocaena 0.2-0.5 sec. In Lipotes, a short dive would take 10-20 sec and a long dive up to 1-2 min (average value being 31 sec). In Neophocaena a long dive would reach a maximum of 65 sec with an average of 17.5 sec. Lipotes has a pair of functional eyes. The selective secondary atrophy of a number of eye muscles and nerves and the position of the eyes in the head correspond with the physical conditions of the environments, so that the field of vision is directed forwards and upwards. When pursued by motor boat, both species react with a long dive and a change of direction under water. With the Baiji this would occasionally cause fatal injuries by the propeller of the ship. Such a case seems much less often with Neophocaena. The population of Lipotes is at present very small. In the stretch of the river (between Nanjing and Taiyangzhou) under exploration, we found scarcely more than one dolphin per 4 km. The cause for its scarcity is mostly due to accidents or wounds caused by propellers.

Adaptation, Physiological↗