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Biomedical subjects

H C Chang

Publications and source records attributed to H C Chang.

At least 127 records · Page 7Linked to original sources
PubMed1997

Affinity and kinetics of the interactions between an alphabeta T-cell receptor and its superantigen and class II-MHC/peptide ligands.

Immune activation is mediated by a specific interaction between the T-cell receptor (TCR) and an antigenic peptide bound to the major histocompatibility complex (MHC). T-cell activation can also be stimulated by superantigens which bind to germline-encoded variable domain sequences of certain TCR beta-chains. We have used a surface plasmon resonance biosensor to characterize the molecular interactions between a class II-restricted alphabeta TCR and its superantigen and MHC/peptide ligands. The extracellular domains of the murine D10 TCR (Valpha2, Vbeta8.2) were expressed in insect cells and secreted as a disulfide-linked heterodimer. In the absence of MHC class II, purified soluble D10 TCR bound to Staphylococcus aureus enterotoxin C2 with an association rate of 1.69+/-0.12 x 10(4)M(-1) sec(-1) and a dissociation rate of 1.9+/-0.47 x 10(-2) sec(-1), giving a dissociation constant of 1.1 microM. Binding of the TCR to S. aureus enterotoxin B was barely detectable and could not be measured accurately due to the rapid dissociation rate. Soluble D10 TCR also bound to a soluble murine MHC class II I-A(k) molecule containing a fused antigenic conalbumin peptide and complementary leucine zipper sequences to facilitate efficient chain pairing. The purified I A(k) chimera specifically stimulated proliferation of the D10 T-cell clone, and bound to immobilized soluble D10 TCR with an association rate of 1.07+/-0.19 x 10(4)M(-1)sec(-1) and a dissociation rate of 2.2+/-0.65 x 10(-2) sec(-1), giving a dissociation constant of 2.1 microM.

Animals↗
PubMed1997

Particle-bound PAH content in ambient air.

Ambient air samples from a traffic intersection, an urban site and a petrochemical-industrial site (PCI) were collected by using several dry deposition plates, two Microorifice uniform deposited impactors (MOUDIs), one Noll Rotary Impactor (NRI) and several PS-1 (General Metal Work) samplers from March 1994 to June 1995 in southern Taiwan, to characterize the atmospheric particle-bound PAH content of these three areas. Twenty-one individual polycyclic aromatic hydrocarbons (PAHs) were analyzed primarily by using a gas chromatograph/mass spectrometer (GC/MS). In general, the sub-micron particles have a higher PAH content. This is due to the fact that soot from combustion sources consists primarily of fine particles and has a high PAH content. In addition, a smaller particle has a higher specific surface area and therefore may contain more organic carbon, which allows for more PAH adsorption. For a particle size range between 0.31 and 3.2 microm, both Urban/Traffic and PCI/Traffic ratios of particle-bound total-PAH content have the lowest values, ranging from 0.25 to 0.28 (mean = 0.26) and from 0.07 to 0.13 (mean = 0.10), respectively. This indicates that, during the accumulation process, the PAH mass shifted from a particle phase to a gas phase, or the particles aggregated with lower PAH-content particles, resulting in a reduction in particle-bound PAH content. By using the particle size distribution data, the dry deposition model in this study can provide a good prediction for the PAH content of dry deposition materials. In general, lower molecular weight PAHs had a larger fraction of dry deposition flux contributed by the gas phase; for 2-ring PAH (50.4, 46.3 and 28.4%), 3-ring PAHs (15.2, 15.4 and 11.7%) and 4-ring PAHs (13.0, 3.60 and 5.01%) for the traffic intersection, urban and PCI sites, respectively. For higher molecular weight PAHs-5-ring, 6-ring and 7-ring PAHs-their cumulation fraction (F%) of dry deposition flux contributed by the gas phase was lower than 3.26%. At the traffic intersection, urban and PCI sites, the mass median diameter of dry deposition materials (MMD(F)) of individual PAHs was between 25.3 and 49.6 microm, between 27.6 and 43.9 microm, and between 19.1 and 41.9 microm, respectively. This is due to the fact that PAH dry-deposition primarily resulted from gravitational settling of the coarse particulates (> 10 microm).

Journal Article↗
PubMed1997

Regiospecificity of peroxyl radical addition to (E)-retinoic acid.

The regiochemistry of peroxyl radical addition to (E)-retinoic acid (RA) was investigated. Peroxyl radicals, generated by reaction of 13-hydroperoxy-(9Z,11E)-octadecadienoic acid with hydroxo(porphyrinato)iron(III) in Tween 20 micelles, were reacted with RA. The major, and virtually exclusive, RA oxidation product was 5,6-epoxy-RA which was identified on the basis of cochromatography with the synthetic synthetic oxirane (in a reverse phase HPLC system), electronic absorption spectroscopy, high-field 1H-NMR, and EI mass spectrometry. These results suggest that peroxyl radicals react with RA by regioselective addition to either C5 or C6 yielding an endocyclic tertiary allylic or tertiary carbon-centered radical adduct, respectively. Subsequent beta-elimination of an alkoxyl radical yields the oxirane. Computational studies were carried out in order to gain mechanistic insights into the observed regiospecificity of the peroxyl radical-dependent epoxidation reaction; molecular mechanics and semiempirical quantum mechanical calculations were carried out using Tripos force field parameters and AM1, respectively. The results suggest that the regiospecific epoxidation may be influenced by the 5,6-olefinic function behaving as a partially-isolated double bond as well as inherent allylic A1,2 strain in the substituted cyclohexene ring as a consequence of substitutions at C1 and C6. In addition, calculated heats of formation indicated preferential peroxyl radical addition to C5 versus C6; this may reflect differences in the geometries of sp2-orbitals containing the radical densities rather than resonance contributions by the highly conjugated polyene system.

Chromatography, High Pressure Liquid↗
PubMed1997

HIV-1 Tat protein exits from cells via a leaderless secretory pathway and binds to extracellular matrix-associated heparan sulfate proteoglycans through its basic region.

OBJECTIVE: To analyze the mechanisms of release and the extracellular fate of the HIV-1 Tat protein and to determine the Tat domain binding to the extracellular matrix. DESIGN AND METHODS: Release of Tat was studied by pulse-chase experiments with Tat-transfected COS-1 cells in the presence or absence of different serum concentrations, temperatures and drugs inhibiting the classical secretion pathway or endo-exocytosis, such as brefeldin A and methylamine. The binding of extracellular Tat to heparan sulfate proteoglycans (HSPG) was determined by using trypsin, heparin or heparinase in pulse-chase experiments, by gel shift and competition assays with radiolabeled heparin, and by heparin-affinity chromatography. The mapping of the Tat binding site to heparin was defined by functional assays of rescue of Tat-defective HIV-1 proviruses. RESULTS: Tat is released in the absence of cell death or permeability changes. Tat release is dependent upon the temperature and serum concentration, and it is not blocked by brefeldin A or methylamine. After release, a portion of the protein remains in a soluble form whereas the other binds to extracellular matrix (ECM)-associated HSPG. The HSPG-bound Tat can be retrieved into a soluble form by heparin, heparinase or trypsin. Binding to heparin is competed out by heparin-binding factors such as basic fibroblast growth factor (bFGF), and it is mediated by the Tat basic region which forms a specific complex with heparin which blocks HIV-1 rescue by exogenous Tat and allows purification of a highly biologically active protein. CONCLUSIONS: These results demonstrate that Tat exits from intact cells through a leaderless secretion pathway which shares several features with that of acid FGF or bFGF. The released Tat binds to HSPG through its basic region and this determines its storage into the ECM, as occurs for bFGF.

Animals↗
PubMed1997

In vivo analysis of the Hsp90 cochaperone Sti1 (p60).

Hsp90 interacts with Sti1 (p60) in lysates of yeast and vertebrate cells. Here we provide the first analysis of their interaction in vivo. Saccharomyces cerevisiae mutations that eliminate Sti1 or reduce intracellular concentrations of Hsp90 individually have little or no effect on growth at normal temperatures. However, when combined, the mutations greatly reduce or eliminate growth. Furthermore, overexpression of Sti1 has allele-specific effects on cells carrying various hsp90ts point mutations. These genetic interactions provide strong evidence that Hsp90 and Sti1 interact in vivo and that their functions are closely allied. Indeed, deletion of STI1 reduces the in vivo activity of the Hsp90 target protein, glucocorticoid receptor (GR). Mutations in GR that eliminate interaction with Hsp90 also eliminate the effects of the sti1 deletion. Examination of GR protein complexes in the sti1 deletion mutant reveals a selective increase in the concentration of GR-Ydj1 complexes, supporting previous hypotheses that Ydj1 functions at an early step in the maturation of GR and that Sti1 acts at an intermediate step. Deletion of STI1 also reduces the in vivo activity of another, unrelated Hsp90 target protein, v-Src. Our data indicate that Sti1 is a general factor in the maturation of Hsp90 target proteins and support earlier suggestions that Hsp90 matures even very different target proteins by a similar mechanism.

Amino Acid Isomerases↗
PubMed1997

Clinical experience of HIV/AIDS in a municipal hospital in Taiwan.

To describe the spectrum of epidemiological and major clinical manifestations of patients infected by human immunodeficiency virus type 1 (HIV-1) in a municipal hospital, a retrospective review was done of 53 HIV-1-infected patients who had been admitted to Taipei Municipal Jen-Ai Hospital between January 1990, and July 1996. The majority (94.3%) of the patients in the cohort were male. Peak incidence was found in the fourth decade (28.3%). Forty-four (83%) patients presented in the first hospital stay with acquired immunodeficiency syndrome (AIDS). The mean duration between establishment of diagnosis of HIV-1 infection and that of AIDS was 11.2 (0-84) months. Heterosexual transmission accounted for 54.7% of the infections in the study group, and bi-/homosexual men made up another 32%. Psychosis of new onset was noted in two patients. In all AIDS indicator conditions, Pneumocystis carinii pneumonia (PCP) was the leading opportunistic infection among AIDS patients. PCP was also on the top of initial manifestations of HIV-1 infection. One patient with Penicillium marneffei infection was diagnosed to have AIDS. The mean CD4 count at admission of AIDS patients was much lower than that of non-AIDS patients (32 vs. 297/microliter, p < 0.0005). During the follow-up period 24 of 53 patients died. Mean survival time of 23 expired patients after establishment of diagnosis of AIDS was 6.4 (0-29) months. The results indicated that males outnumbered females greatly in the number of cases. Sexual activity remained the most important route of infection. Psychosis of new onset may be an early manifestation of HIV-associated encephalopathy and requires more attention. In addition, the outcome was poor as most patients in this area did not become aware of risk of HIV-1 infection until they were seriously illed with full-blown AIDS that they would seek medical help. PCP was the most common incentive for medical consultation. Penicillium marneffei infection is endemic in southeast Asia, and should be classified as an AIDS indicator condition in Taiwan.

AIDS-Related Opportunistic Infections↗
PubMed1997

Reflux esophagitis after proximal subtotal gastrectomy.

BACKGROUND: Carcinoma of the cardia of stomach or carcinoma of the esophagocardiac (EC) junction has a poor prognosis. Proximal subtotal gastrectomy is often performed for a smaller tumor. It achieves a prognosis similar to that of total gastrectomy and prevents pernicious anemia. A better functional result was expected than that total gastrectomy did from us, but the clinical observation was surprising. Most patients had symptoms of reflux esophagitis after surgery. All of these patients were reevaluated. METHODS: From November, 1990 to March, 1996, there were 27 patients who received proximal subtotal gastrectomy at this hospital. Twenty of these were involved in this study. Floppy Nissen's fundoplication was performed during operation as a anti-reflux procedure. Continuous 24-h pH monitoring, esophageal manometry, and endoscopic examination were performed for study. RESULTS: Most of the patients had symptoms of gastroesophageal reflux. 80% (16/20) of the patients were found to have reflux esophagitis under endoscopy. The patients had higher pH scores (mean 85.8 +/- 45.6) than normal (< 17.5), lower pressure of LES (8.03 +/- 7.01 mmHg), shorter total lengths (mean 1.60 +/- 0.56 cm) and intraabdominal lengths (mean 1.29 +/- 0.30 cm) of new lower esophageal sphincters. CONCLUSIONS: Clearly there was failure to prevent reflux esophagitis through floppy Nissen's fundoplication. Severe gastroesophageal reflux did appear after proximal gastric resection. A gastric remnant with safe cut-margin is often too short to perform complete wrapping. Proximal subtotal gastrectomy is not recommended for early cancer of the cardia of the stomach.

Aged↗
PubMed1997

Identification and differentiation of bear bile used in medicinal products in Taiwan.

One hundred eighty-three suspect bear bile used in medicinal products, collected in Taiwan as gall bladders or dried powder forms, were analyzed using FTIR, HPTLC, and HPLC techniques to identify whether they are indeed bear bile. Those confirmed were further examined to determine whether the observed analytical parameters can be reliably used for source inference, i.e., differentiating products among North American black bear, farmed Asiatic black bear, polar bear, etc. Our data suggested that North American and polar bears contain a higher concentration of TC (relative to TUDC and TCDC), whereas the relative concentration of TC in Asiatic bears (wild or farmed) is much lower. Thus, the relative concentration of TC can potentially be used for differentiating Asiatic bear bile from North American and polar bear products, but it cannot be used for the differentiation of wild and farmed bear bile as suggested in an earlier report by Espinoza et al. The origin of the 183 samples analyzed were found to be as follows: 118 (64%), bile salts, or gall bladders were of domestic pig; 56 (31%), bile products of Asiatic bear; 4 (2.2%), Asiatic bear mixed with pig bile salts; 3 (1.6%) goat gall bladders; 1 (0.55%) water buffalo bile salts; and 1 (0.55%), pig bile salts mixed with water buffalo bile salts.

Animals↗
PubMed1996

Crystallization of a deglycosylated T cell receptor (TCR) complexed with an anti-TCR Fab fragment.

A strategy to overexpress T cell receptors (TCRs) in Lec3.2.8.1 cells has been developed using the "Velcro" leucine zipper sequence to facilitate alpha-beta pairing. Upon secretion in culture media, the VSV-8-specific/H2-Kb-restricted N15 TCR could be readily immunopurified using the anti-leucine zipper monoclonal antibody 2H11, with a yield of 5-10 mg/liter. Mass spectrometry analysis revealed that all attached glycans were GlcNAc2-Man5. Following Superdex 200 gel filtration to remove aggregates, wild-type N15 or N15(s), a C183S variant lacking the unpaired cysteine at amino acid residue 183 in the Cbeta domain, was thrombin-cleaved and endoglycosidase H-digested, and the two derivatives were termed iN15DeltaH and N15(s)DeltaH, respectively, and sized by Superdex 75 chromatography to high purity. N-terminal and C-terminal microsequencing analysis showed the expected unique termini of N15 alpha and beta subunits. Nevertheless, neither protein crystallized under a wide range of conditions. Subsequently, we produced a Fab fragment of the murine TCR Cbeta-specific hamster monoclonal antibody H57 and complexed the Fab fragment with iN15DeltaH and N15(s)DeltaH. Both N15(s)DeltaH-Fab[H57] and iN15DeltaH-Fab[H57] complexes crystallize, with the former diffracting to 2.8-A resolution. These findings show that neither intact glycans nor the conserved and partially exposed Cys-183 is required for protein stability. Furthermore, our results suggest that the H57 Fab fragment aids in the crystallization of TCRs by altering their molecular surface and/or stabilizing inherent conformational mobility.

Animals↗
PubMed1996

A cyclophilin function in Hsp90-dependent signal transduction.

Cpr6 and Cpr7, the Saccharomyces cerevisiae homologs of cyclophilin-40 (CyP-40), were shown to form complexes with Hsp90, a protein chaperone that functions in several signal transduction pathways. Deletion of CPR7 caused severe growth defects when combined with mutations that decrease the amount of Hsp90 or Sti1, another component of the Hsp90 chaperone machinery. The activities of two heterologous Hsp90-dependent signal transducers expressed in yeast, glucocorticoid receptor and pp60(v-src) kinase, were adversely affected by cpr7 null mutations. These results suggest that CyP-40 cyclophilins play a general role in Hsp90-dependent signal transduction pathways under normal growth conditions.

Amino Acid Isomerases↗
PubMed1996

Characterization of the replicon of plasmid pSW500 of Erwinia stewartii.

A 1.6-kb DNA region required for the replication of pSW500 from Erwinia stewartii SW2 has been identified. DNA sequencing analysis revealed that this DNA fragment consists of a DnaA box, seven 16-bp direct repeats, and a 1005-bp open reading frame. The seven direct repeats have been demonstrated to mediate the incompatibility function of the plasmid. Primer extension analysis showed that the 1005-bp ORF is transcribed in vivo and the +1 site of the transcript is located 113 bp upstream from the translation initiation codon of the ORF. Complementation studies showed that this ORF is required for the replication of the plasmid and may encode a replication protein, RepA. Gene fusion studies revealed that the expression of repA is autoregulated by RepA. We also found that the pSW500 replicon has a copy number of approximately two and that the plasmid is stably maintained in Escherichia coli, thus demonstrating that the replicon contains all the elements required for copy number control and plasmid stability in E. coli. Curing of pSW500 from E. stewartii SW2 revealed that loss of pSW500 did not have any obvious effect on morphology or physiology of the cells, suggesting that pSW500 does not encode a function that is indispensable for the survival of the organism.

Amino Acid Sequence↗
PubMed1996

Nonenzymatic reduction of tetrachloro-1, 4-benzoquinone by reduced nicotinamide adenine dinucleotide phosphate in an aqueous system.

NADPH was shown to reduce tetrachloro-1,4-benzoquinone (TCBQ) to tetrachloro-1,4-benzene diol which was identified on the basis of cochromatography with synthetic standard in a reverse phase HPLC system, electronic absorption spectroscopy, and mass spectrometry. Conversely, NADPH was shown to undergo TCBQ-dependent oxidation as evidenced by uv spectroscopic analysis. ESR spectroscopy demonstrated that NADPH-dependent reduction of TCBQ proceeds through the intermediacy of a semiquinone radical intermediate. In addition, molecular mechanics and molecular orbital calculations were carried out on TCBQ to obtain partial atomic charges and the results suggest that the first electron transfer from the reduced nicotinamide moiety to TCBQ occurs at a carbonyl carbon. Kinetic analysis indicated that the rate of NADPH oxidation is first order with respect to [TCBQ], second order with respect to [NADPH], and an Arrhenius plot yielded an activation energy of 26 kJ x mol (-1) for the rate limiting step. Mechanisms are proposed which are consistent with these results.

Chloranil↗
PubMed1996

Protein phosphatase 2A positively and negatively regulates Ras1-mediated photoreceptor development in Drosophila.

Protein phosphatase 2A (PP2A), a heterotrimeric serine/threonine phosphatase present in most tissues and cell types, has been implicated in the regulation of cell cycle progression, DNA replication, transcription, and translation. Here we present genetic evidence suggesting that PP2A functions downstream of Ras1 in the Sevenless receptor tyrosine kinase (RTK) signal transduction pathway that specifies R7 photoreceptor cell fate in the developing Drosophila eye. Ras1 and downstream cytoplasmic kinases, Raf, MEK, and MAPK, comprise an evolutionarily conserved cascade that mediates the transmission of signals from RTKs at the plasma membrane to specific factors in the nucleus. Using transgenic flies expressing constitutively activated Ras1 or Raf proteins that function independently of upstream signaling events, we show that a reduction in the dose of the gene encoding the catalytic subunit of PP2A stimulates signaling from Ras1 but impairs signaling from Raf. This suggests that PP2A both negatively and positively regulates the Ras1 cascade by dephosphorylating factors that function at different steps in the cascade.

Animals↗
PubMed1996

Okadaic acid, sphingosine, and phorbol ester reversibly modulate heat induction on protein kinase FA/GSK-3 alpha in A431 cells.

Exposure of A431 cells to a rapid and sudden increase from 37 degrees C to 46 degrees C for 30 min could induce an increase in protein level and cellular activity of protein (kinase FA/GSK-3 alpha) up to approximately 200% of control level. However, when cells were first treated with 500 nM tumor promoter phorbol ester TPA at 37 degrees C for 30 min to activate cellular protein kinase C (PKC) or with 400 nM okadaic acid at 37 degrees C for 30 min to inhibit cellular protein phosphatases followed by heat shock at 46 degrees C for another 30 min, the heat induction on kinase FA/GSK-3 alpha was found to be completed blocked. In sharp contrast, when cells were first treated with 1 microM TPA at 37 degrees C for 24 h or with 5 microM sphingosine at 37 degrees C for 30 min to down-regulate cellular PKC, the heat induction on kinase FA/GSK-3 alpha was found to be reversely promoted up to approximately 250% of control level, demonstrating that kinase FA/GSK-3 alpha may not represent a constitutively active/mitogen-inactivated protein kinase as previously conceived. Taken together, the results provide initial evidence that TPA/sphingosine and okadaic acid could reversibly modulate the heat induction on kinase FA/GSK-3 alpha in A431 cells, suggesting that phosphorylation/dephosphorylation mechanisms are involved in the regulation of the heat-shock induction of kinase FA/GSK-3 alpha, representing a new mode of signal transduction for the regulation of this multisubstrate protein kinase and a new mode of signaling pathway modulating the heat-induction process.

Amino Acid Sequence↗
PubMed1996

A prospective trial of tacrolimus (FK 506) in clinical heart transplantation: intermediate-term results.

Between January 1, 1989, and December 31, 1994, we have treated 122 primary heart recipients with FK 506 (group I) and 121 with cyclosporine (group II). Fifty patients in the cyclosporine (CyA) group received no lympholytic induction (CyA alone) and 71 others received lympholytic induction with either rabbit antithymocyte globulin or OKT3 (CyA+LI). The mean follow-up was longer in the FK 506 group than in the CyA groups (3.2 +/- 1.3 vs 2.3 +/- 1.8 years; p< 0.01). Patient survival did not differ on the basis of the type of immunosuppression used. At 3 months after transplantation, the freedom from rejection in the FK 506 group was higher than that of the CyA-alone group (47% vs 22%, p < 0.01) but similar to that of the CyA+LI group (47% vs 53%). The linearized rejection rate (episodes/100 patient-days) of the FK 506 group (0.09 episodes) was lower (p < 0.05) than that of the CyA-alone group (0.26) and the CyA+LI group (0.13). The requirement for pulsed steroids to treat rejection was less in common in the FK 506 group than in either CyA group. Eighteen patients in the CyA group had refractory rejections; all resolved with FK 506 rescue. Two patients in the FK 506 group had refractory rejection that resolved with total lymphoid irradiation (n=1) and methotrexate therapy (n=1). Patients receiving FK 506 had a lower risk of hypertension and required a lower dose of steroids. Although the mean serum creatinine concentration at 1 year was higher in the FK 506 group, this difference disappeared after 2 years. No patients required discontinuation of FK 506 because of its side effects. Our intermediate-term results indicate that FK 506 compares favorably with CyA as a primary immunosuppressant in heart transplantation.

Adult↗
PubMed1996

A screen for genes that function downstream of Ras1 during Drosophila eye development.

Cell-fate specification of the R7 photoreceptor cell is controlled by the sevenless receptor tyrosine kinase (SevRTK) and Ras1, the Drosophila homologue of mammalian H-ras, K-ras and N-ras oncogenes. An activated form of Ras1 expressed under control of the sevenless enhancer/promoter (sev-Ras1V12) induces production of supernumerary R7 photoreceptor cells, which causes the eye to become rough in appearance. To isolate mutations in genes functioning downstream of Ras1, we carried out a screen for dominant suppressors and enhancers of this rough eye phenotype. Approximately 850,000 mutagenized flies were screened, and 282 dominant suppressors and 577 dominant enhancers were isolated. Mutations in the Drosophila homologues of Raf, MEK, MAPK, type I Geranylgeranyl Transferase and Protein Phosphatase 2A were isolated, as were mutations in several novel signaling genes. Some of these mutant genes appear to be general signaling factors that function in other Ras1 pathways, while one seems to be more specific for photoreceptor development. At least two suppressors appear to function either between Ras1 and Raf or in parallel to Raf.

Animals↗
PubMed1996

Pulse spectrum analysis of chemical factory workers with abnormal blood test.

Double blind tests to check the correlation between pulse diagnosis and liver function tests were performed. Blood tests including T-Bil, D-Bil, SGOT and SGPT of 70 chemical factory workers were compared with pulse analysis. Special attention was paid to the indicators of liver and lung meridians for pulse diagnosis. It was found that using the criterion (1) C1 > or = 3+ and C1 + C4 > or = 4+ (hyperfunction) and (2) C1 < or = 3-(hypofunction) as abnormal liver meridian (for C1, every 5% above normal give one +, every 5% below normal give one-; for C4 every 10% above normal give one +, every 10% below normal give one -). The correlation or agreement between the blood tests and the pulse diagnosis was very high (noncorrelation chance checked by X2-test, P < 0.001, degree of agreement checked by Kappa test Ka = 0.61 which means a substantial relationship). Suggestions such as more tests, more criteria and precautions for future study are also proposed.

Adult↗