Search

In order to elucidate the roles of 17β-HSDs in fish gonadal steroidogenesis, three types of 17β-HSDs (17β-HSD1, 17β-HSD8 and putative 17β-HSD12) were cloned and characterized from the Nile tilapia, Oreochromis niloticus. The cloned cDNAs of 17β-HSD type 1, 8 and 12 were 1504, 1006 and 1930 bp long, with open reading frames encoding proteins of 289, 256 and 314 aminoacids, respectively. Tissue distribution pattern analyzed by RT-PCR and Northern blot showed that 17β-HSD1 was dominantly expressed in the ovary, while the putative 17β-HSD12, one of the two duplicates found in fish, is a male specific enzyme and expressed exclusively in testis (detected by RT-PCR only). On the other hand, 17β-HSD8 was expressed in the brain, gill, heart, liver, intestine, gonad, kidney and muscle of both male and female. Enzymatic assays of the three types of 17β-HSDs were performed using recombinant proteins expressed in E. coli or HEK 293 cells. Tilapia 17β-HSD1 expressed in E. coli had the preference for NADP(H) as cofactor and could catalyze the inter-conversion between estrone and estradiol efficiently as well as the inter-conversion between androstenedione and testosterone, but less efficiently. Tilapia 17β-HSD8 recombinant protein expressed in HEK 293 cells could catalyze the conversion of testosterone to androstenedione, as well as the inter-conversion between estrone and estradiol. However, the putative 17β-HSD12 expressed in E. coli or in HEK 293 cells showed no conversion to any of the four substrates tested in this study. Based on enzyme characterization and tissue distribution, it is plausible to attribute crucial roles to 17β-HSDs in the gonadal steroidogenesis of teleosts.

Abnormal proliferation of vascular smooth muscle cells (VSMCs) is known to be a key event in the development of atherosclerosis and restenosis. The present study examined the effect of a novel synthetic natriuretic peptide, vasonatrin peptide (VNP), on norepinephrine (NE)-induced proliferation of VSMCs from coronary bypass vessels. Human VSMCs were isolated from an internal mammary artery (IMA) and saphenous vein (SV) by explant culture and stimulated with NE. MTT assay and [3H] thymidine-incorporation were undertaken to analyze cell proliferation and radioimmunoassay was used to determine the level of intracellular cyclic 3’,5’-guanosine monophosphate (cyclic GMP). NE (10-8 - 10-7 mol/l) had a mitogenic effect in human VSMCs from both SV and IMA. However, NE-stimulated proliferation of VSMCs from SV was greater than that from IMA. Furthermore, low concentration of NE (10-10 mol/l) promoted cell growth in SV-derived cells but not in IMA-derived cells. VNP (10-8 - 10-6 mol/l) reduced NE-induced cell proliferation and increased intracellular cyclic GMP, which were abrogated by HS-142-1. In addition, the growth inhibition of VNP was mimicked by 8-bromo-cGMP. These results indicate that VNP has a significant inhibitory effect on NE-stimulated proliferation of human VSMCs from both IMA and SV, which is mediated by guanylate cyclase-linked receptors by increasing cyclic GMP.

Interleukin-6 (IL-6), a pleiotropic cytokine, is postulated to be involved in the pathogenesis of sick euthyroid syndrome, although the direct in vitro effects of IL-6 on human thyroid function are controversial. Because IL-6 signal can be transduced when the complex of IL-6 and soluble IL-6 receptor (sIL-6R) binds to gp 130, an IL-6 signal transducer, we studied the effects of IL-6 and sIL-6R on thyroid function, using human thyroid follicles obtained from patients with Graves' disease. IL-6 alone had no inhibitory effect on TSH-induced thyroid function (125I incorporation and organic 125I release), even at supraphysiological concentrations. However, in the presence of physiological concentrations of sIL-6R (100 ng/ml), IL-6 inhibited thyroid function dose dependently and completely, accompanied with the decreased ratio of 125I-T3/125I-T4 not only in the thyroid follicles but also in the culture medium. Thyroid follicles did not secrete sIL-6R but produced IL-6 constitutively. Consistent with these findings, sIL-6R inhibited thyroid function slightly at high concentrations. Furthermore, RT-PCR analyses revealed that human thyroid follicles expressed the messenger RNAs for IL-6 and gp130 but scarcely messenger RNA for IL-6R. These in vitro findings suggest that IL-6 alone hardly affects thyroid function in thyroid follicles in which IL-6R gene is scarcely expressed. However, because sIL-6R is present abundantly in serum, IL-6 in vivo would be capable of inhibiting the synthesis and release of T4 and, to a greater extent, T3 from the thyroid gland. These in vitro findings are at least partly related to the development of sick euthyroid syndrome.

A special local burn wound treatment, using a healing promotion factor, attempted to achieve earlier wound closure. Clinically it is not difficult to harvest autologous human platelet-derived wound healing factor (HPDWHF) from burn patients. Another study has proved that porcine pituitary extract (PPE) could be one of the supplements for growing human keratinocytes in vitro (Wang HJ, Chen TM, Tung YM et al. Burns 1995). Initially, using the ELISA immunoassay, we detected titres of platelet-derived growth factor heterodimer AB (PDGF-AB) and epidermal growth factor (EGF) in HPDWHF and PPE. After elevating a 6 X 9 cm2 flap on the backs of Sprague-Dawley (SD) rats 1 ml of heterologous HPDWHF and PPE were sprayed topically on the wound and followed by grafting six pieces of 1 cm2 autologous full-thickness skin. The flap wound was then closed with a piece of large-pore Biobrane interposed between the skin graft wound and the flap. On postgraft day 7, the wound was reopened to measure the area of the full-thickness skin graft (FTSG) using micrographic paper; the results are reported as the percentage of graft expansion beyond the original size. In the cytokine study, we found that the concentrations of PDGF-AB dimer in HPDWHF and PPE were 5222 +/- 102 and 375 +/- 12 pg/ml (both values are reported after a 1:10 dilution) (n = 3), whereas the EGF concentration was 245 +/- 9 pg/ml, and undetectable in undiluted PPE (n = 3), thereby proving that the PDGF-AB and EGF fibres in the HPDWHF were significantly higher than in the PPE Both the HPDWHF (n = 6, with each animal grafted with six skin graft samples) and PPE (n = 7) demonstrated significant accelerations of FTSG growth, with 14.41 +/- 1.08 per cent and 13.16 +/- 3.25 per cent increases in the FTSG size when compared to the sham control group 1.39 +/- 3.26 per cent (n = 5) (P0.05). Comparisons between the treatment groups showed no evidence to indicate that the HPDWHF is superior to the PPE in accelerating FTSG growth (P0.05). This supports the hypothesis that PDGF appears to transduce its signal through wound macrophages and may trigger the induction of positive autocrine feedback loops and synthesis of endogenous wound PDGF and other growth factors, thereby enhancing the cascade of tissue repair (Pierce GF, Mustoe TA, Altrock B et al. 1991; J Cell Biochem 1991; 45: 319-326), and the effect that PDGF-AB on wound healing is not dose dependent (Mustoe TA, Cutler NR, Allman RM et al. Arch Surg 1994; 129: 213-219). In future studies we expect to show that autologous HPDWHF and heterologous PPE will promote skin graft wound healing in burn patients since they contain a high concentration of PDGF-AB.

We evaluated the putative factors for the onset of Guillain-Barre syndrome (GBS) in Harbin, China by a case-control study based on the information from GBS patients identified from a population-based incidence survey, which is the first study of this kind in China. Sixty-nine GBS patients were identified during a 1-year period from 1 October 1997 to 30 September 1998, and they were matched with 69 controls for gender and age (+/-5 years). GBS diagnosis was validated by senior neurologists and GBS patients were followed up for 6 months after onset. Odds ratio (OR) and 95% confidence intervals (CI) for each putative factor for the onset of GBS were calculated and compared between GBS cases and controls. Precedent respiratory infections within 2 months before onset were found to be significantly more frequent in GBS patients than in controls (OR = 1.68, 95% CI: 1.21-2.33). Although the number of cases with gastroenteritis among GBS patients was more than double of that in the controls, the difference was not statistically significant (OR = 2.25, 95% CI: 0.73-6.96). Other putative factors as well as characteristics regarding family situation, education level, occupation, etc., were not found to be statistically different between GBS patients and controls.

The authors carried out a door-to-door survey to determine the prevalence of epilepsy among 55,000 people in China. The lifetime prevalence was 7.0/1000, and 41% of all persons had never received appropriate treatment. The prevalence of active epilepsy was 4.6/1000, and 63% of people with active epilepsy had not received antiepileptic treatment in the week before the survey. Figures for the prevalence and the treatment gap were significantly higher than previous estimates.

To investigate and compare the common effect of Bailong, a Chinese herbal preparation, and Hexamethylene bisacetamide (HMBA), a western cell differentiation inducer, on growth regulation and phenotype of human gastric cancer (MGC80-3) cells in different phases of cell cycle.Synchronized MGC80-3 cells in monolayer cultures with nitrous oxide under high pressure were double blocked with overdosage of TdR in the 4 cellular phases (G1, S, G2 and M), and then collected and treated with Bailong and 5 mmol/L HMBA respectively.Both medicines could suppress the cell proliferation, soft agar cloning growth and microfilament assembling of different phases of cells, the effect on cells of G1 phase was the most significant one.Bailong and HMBA had significant common characteristics in inducing cell differentiation of MGC80-3 cells in different phases of cell cycle.

To investigate the common regulative effects of the Chinese drug Bailong and hexamethylen bisacetamide (HMBA) on expressions of oncogenes (c-H-ras and c-myc), and tumor suppressor genes (Rb, p53 and p21) of MGC80-3 in human cancer cell cycle.Adopting RNA Northern Blot to survey the levels of gene expressions of MGC80-3 different phases cells treated with Bailong and HMBA respectively.In different phases of MGC80-3 cells treated with Bailong and differentiation inducer HMBA, expressions of oncogenes c-H-ras and c-myc were inhibited by over 50.0%, messenger kinase subspecies PKC-alpha gene is similar with the expression inhibition of oncogenes, except effect of Bailong on the G2 phase in cell cycle. Effect of Bailong differs greatly from HMBA in the expression of tumor suppression genes. The expression of Rb and p21 in cells treated by HMBA did not increase but were inhibited by 39.5% and 33.3% respectively in G1 phase. The level of Rb gene expression was decreased, too by 3.0% in S phase. Comparison with HMBA the expression of Rb and p21 genes were increased after treatment by Bailong in all cell cycle. But the effect of Bailong on the expression of p53 gene which was increased obviously by 125.0%-233.4% in majority phase of MGC80-3 cells is similar to HMBA.(1) The effect of Bailong on the regulation of oncogenes and tumor suppressor gene is similar to HMBA but the effect of Bailong is better than that of HMBA. (2) Molecular mechanism of the Bailong or HMBA on the proliferative inhibition and differentiation of MGC80-3 related to regulation of the Bailong and HMBA on the oncogenes and tumor suppressor genes in cell cycle of MGC80-3.

To study the features of cell death induced by the anticancer antibiotic lidamycin (LDM) in human hepatoma BEL-7402 cells.Chromatin condensation was observed by co-staining with fluorescent dyes, hoechst 33342 and propidium iodide. "G1 sub-peak" was detected by flow cytometry and DNA ladder was observed using agarose gel electrophoresis. The caspase-3, 6 activities were measured with kits specific for them.Typical apoptotic chromatin condensations appeared when the BEL-7402 cells were treated with the conventional antitumor agent mitomycin C30 mumol.L-1 for 12 h. However, an abnormal type of chromatin condensation occurred when the cells were treated with LDM 1 mumol.L-1 for 6 h, which was characterized with keeping the completeness of nuclear membrane and not forming apoptotic bodies. The DNA ladder patterns were observed using agarose gel electrophoresis. The "G1 sub-peak" occurred only in the cells treated with LDM for 24 h, though chromatin condensation was earlier detected in treatment with LDM for 6 h. The caspase-3, 6 activities were increased about 5 and 4 folds, after the cells were treated with LDM 1 mumol.L-1 for 6 h, as did mitomycin C. The time of initiating chromatin condensation was earlier than that of the high peak activities of caspase-6.The characterization of cell death induced by lidamycin in the human hepatoma BEL-7402 cells differs from typical apoptosis. The results make it helpful to explain the molecular mechanism of the highly potent cytotoxicities of lidamycin toward tumor cells.

To study the regulation of bailong, a Chinese herbal anticancer preparation and hexamethylene bisacetamide (HMBA) on cyclin dependent kinase inhibitor P16 (CKI-P16) genes in human cancer cells.The expression of CKI-P16 in different human cancer cells treated by Bailong or HMBA under different condition, was examined using Northern hybridization, Western blotting assay, etc.After being treated by Bailong or HMBA, the P16 expression increased. This effect was closely related to co-regulation of cAMP-PKA and DAC-PKC signal pathway. When PKA pathway was blocked with PKA inhibitor, the P16 expression decreased, while PKC pathway was blocked, it enhanced.(1) The low P16 expression in G1 phase of cancer cell, as compared with that in S, G2 and M phases, might be an important factor responsible to the incompetence of P16 in inhibiting effectively the malignant change of cancer cell. (2) Mechanism of Bailong and HMBA on cancer cell proliferation inhibition might be correlated with the enhancement of P16 expression in G1 phase of cancer cells. (3) Regulation and expression of Bailong and HMBA on P16 showed the common character of Chinese and western medicine in regulating cancer cells. (4) This study elucidated that upstream of P16 was related to cAMP/PKC signal pathway closely.

One of the most profound circulatory changes that occur in man during exposure to microgravity is a cephalad redistribution of fluid. Central venous pressure (CVP) is not only an important standard for determining the cephalad redistribution of fluid but also the unique valuable means for measuring continuously cardiac filling pressure under simulated weightlessness (SW) and microgravity, and has a relationship with neurohumoral [correction of nerohumoral] regulation. With the development of manned spaceflight, the researches in space suggested that CVP did not increase, which is directly contradictory to the previous results under SW. This paper mainly discusses the changes of CVP and the effects of countermeasures on CVP under SW and microgravity, and the mechanisms of decreased CVP under microgravity. It is postulated that more suitable SW models, more studies about the area of low pressure including especially pulmonary circulation, and comprehensive studies in multiple systems are needed.

Objective. To accumulate data for further studies on the mechanisms of the decrease of cardiovascular function after weightlessness. Method. Cardiopulmonary circulatory function under supine position and head up tilting (HUT) for 20 min before and after exposure to 7 d 6 degrees head down tilt (HDT) in 6 healthy adults who aged 19-21 were measured with a XXH-2000 lesser (pulmonary) circulation and cardiac function instrument. Result. The 7 d HDT resulted in orthostatic intolerance of the subjects; the reduction of RVET (j-z) and the elevation of RPEP [RVET(Q-j)/(j - z)] indicated a decrease of the right heart function. An obvious decrease of the right heart reserves was also observed under HUT after HDT. Conclusion. hc and hc/hz were demonstrated to be useful parameters to evaluate orthostatic tolerance or to forecast orthostatic syncope. The lesser (pulmonary) circulation and cardiac function testing methods are valuable in evaluating the cardiopulmonary circulatory function during HUT.

Fluid is transferred cephalad in microgravity and simulated microgravity, and the pulmonary circulation is the first to be affected. Blood and fluid contents in the lungs increase, but unevenly distributed among various zones of the lungs. Blood vessels in the lungs are filled and distend. Capillary changes have been observed in the animal model of simulated microgravity. Studies about the changes of regulative function of pulmonary circulation are relatively rare. Observations of the reactivity of pulmonary vessels may help to understand the mechanisms of the changes in pulmonary circulation during microgravity.

Electrocardiography (ECG) is still the only method for continuous medical observation during various stages of foreign manned spaceflight activities, and it also serves as an important item for medical monitoring in the project for manned spaceflight activities in our country. With electrocardiography, pulse or heart rate (one of the four life signs) and cardiac function can continuously be observed. Cardiac phase relative to ECG is a main index reflecting cardiac function. Understanding of the results of ECG and cardiac phase and their mechanisms during various stages of foreign manned spaceflight activities will be helpful in establishing emergency measures in the course of manned spaceflight activities in our country.

Arterial blood pressure (BP) is one of the four life signs in human beings, it could reflect cardiovascular function in some degrees. Thus, BP serves as an important cardiovascular parameter during various stages in foreign spaceflight activities. Although the reports about BP are not always in consistency during spaceflight, hypotension commonly occurs during orthostatic stress after spaceflight. The mechanisms of orthostatic intolerance after spaceflight are virtually the same as that of orthostatic hypotension after spaceflight. This paper simply reviewed the reports on BP during various stages in foreign spaceflight activities and the researches on the mechanism of orthostatic hypotension after spaceflight.

We investigated the incidence of loss of heterozygosity (LOH) and microsatellite instability in sporadic prostate cancer and surrounding tissue at loci encompassing the HPC1 and PTEN genes.Surgical specimens from 63 patients with sporadic stage T3 or T4 prostatic adenocarcinoma were analyzed for LOH and microsatellite instability. Microdissected tissue included morphologically normal foci, benign prostatic hyperplasia (BPH) and prostatic adenocarcinoma. LOH analysis was performed using 4 microsatellite markers that map in the region of the 1q24 to 25 locus of the putative prostate cancer susceptibility gene HPC1 and 4 that map in the region of the 10q23 locus of the PTEN gene.The incidence of LOH on 10q was consistent with that previously reported in prostatic tumors. LOH associated with the PTEN locus was recorded in morphologically normal foci, BPH and adenocarcinoma. Sequence analysis of PTEN in a limited number of lesions revealed mutations in nontumor and tumor tissue. Analysis of the DS10215 locus showed significant LOH in tumor but not in benign tissue, suggestive of a tumor suppressor gene in this region associated with prostatic neoplastic progression. In contrast, no significant LOH was observed in the same tissues at 4 loci on chromosome 1q. In this study we recorded elevated levels of microsatellite instability in benign prostatic tissue with an additional increase associated with prostatic adenocarcinoma.The low incidence of LOH in the region of the HPC1 locus in all prostate lesions studied suggests that this putative hereditary prostate cancer susceptibility locus does not appear to have a role in sporadic prostate cancer, at least not in the context of LOH. In contrast, analysis of the same tissues for LOH at chromosome 10q confirmed frequent alterations in this region linked to late stage prostate cancer. PTEN mutations in microdissected morphologically normal and BPH tissue showed alterations in nontumor tissue surrounding adenocarcinoma. Microsatellite instability was increased in adenocarcinomas over an elevated background recorded in surrounding tissues.

The main differences between physiological effects of microgravity in spaceflight and simulated microgravity on humans appeared in the circulation of the low pressure side, in humoral and electrolyte metabolism. For a further understanding of the physiological effects of microgravity, some improvement of the current human experimental models are needed. It is possible to choose more adequate models closer to the situation in microgravity through measuring cardiovascular parameters, fluid regulation and renal excretion variables under head-up tilt (HUT) plus lower body positive pressure condition, or under head-down tilt (HDT) plus upper body negative pressure in various angles and pressure levels.

Orthostatic intolerance commonly occurs after spaceflight, but its mechanisms remain to be clarified. It is believed that the reduction of blood volume might be one of the important factors. So the current countermeasures against orthostatic intolerance are aimed to control the changes of blood volume. The main differences between physiological effects of microgravity and simulated microgravity on humans appear in the circulation of the low pressure side and in humoral and electrolyte metabolism. Reflexes elicited from the low pressure side are very important for regulation of the extracellular fluids. The authors postulated that for a further understanding of mechanisms of orthostatic intolerance after spaceflight and to establish appropriate countermeasures against orthostatic intolerance, it is important to study the course of changes of the circulation in the low pressure side and the reflexes thus elicited with more appropriate method of simulation.

Loss of heterozygosity (LOH) on 10q is associated with late-stage events in urothelial neoplastic progression. The tumor suppressor gene PTEN, which is mutated or homozygously deleted in numerous cancers, maps to a region of 10q within the reported region of minimal loss in bladder tumors. In two recent studies alterations in the PTEN gene occur at a low frequency in bladder tumors displaying 10q LOH. We have screened 35 late-stage bladder tumors for mutations in PTEN and MXI1, both genes mapping to chromosome 10q. Using single-strand conformation polymorphism analysis, we identified 6 tumors harboring mutations in PTEN and 2 additional tumors displaying homozygous deletion at this locus. No MXI1 mutations were identified within the same tumor panel. Of 16 bladder tumor cell lines analyzed, 2 showed homozygous deletion of PTEN and 3 harbored point mutations resulting in an amino acid change. Two cell lines harbored missense mutations in MXI1. We report a significantly higher frequency of PTEN alterations in bladder carcinoma (23%) than was previously recorded, with no accompanying mutations in the MXI1 gene.

This study presents evidence that phosphoinositide (PI) 3-kinase is involved in T cell Ca(2+) signaling via a phosphatidylinositol 3,4, 5-trisphosphate PI(3,4,5)P(3)-sensitive Ca(2+) entry pathway. First, exogenous PI(3,4,5)P(3) at concentrations close to its physiological levels induces Ca(2+) influx in T cells, whereas PI(3,4)P(2), PI(4, 5)P(2), and PI(3)P have no effect on [Ca(2+)](i). This Ca(2+) entry mechanism is cell type-specific as B cells and a number of cell lines examined do not respond to PI(3,4,5)P(3) stimulation. Second, inhibition of PI 3-kinase by wortmannin and by overexpression of the dominant negative inhibitor Deltap85 suppresses anti-CD3-induced Ca(2+) response, which could be reversed by subsequent exposure to PI(3,4,5)P(3). Third, PI(3,4,5)P(3) is capable of stimulating Ca(2+) efflux from Ca(2+)-loaded plasma membrane vesicles prepared from Jurkat T cells, suggesting that PI(3,4,5)P(3) interacts with a Ca(2+) entry system directly or via a membrane-bound protein. Fourth, although D-myo-inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4, 5)P(4)) mimics PI(3,4,5)P(3) in many aspects of biochemical functions such as membrane binding and Ca(2+) transport, we raise evidence that Ins(1,3,4,5)P(4) does not play a role in anti-CD3- or PI(3,4,5)P(3)-mediated Ca(2+) entry. This PI(3,4,5)P(3)-stimulated Ca(2+) influx connotes physiological significance, considering the pivotal role of PI 3-kinase in the regulation of T cell function. Given that PI 3-kinase and phospholipase C-gamma form multifunctional complexes downstream of many receptor signaling pathways, we hypothesize that PI(3,4,5)P(3)-induced Ca(2+) entry acts concertedly with Ins(1,4,5)P(3)-induced Ca(2+) release in initiating T cell Ca(2+) signaling. By using a biotinylated analog of PI(3,4,5)P(3) as the affinity probe, we have detected several putative PI(3,4,5)P(3)-binding proteins in T cell plasma membranes.

The purpose of this study was to investigate the influence of unilateral condylar neck fracture upon the stress distribution in the mandible and the temporomandibular joint (TMJ) to realize primarily the biomechanical mechanism of the temporomandibular joint disorders (TMD) caused by TMJ injury.The character of stress distribution in the mandible of a normal human and a patient with TMD induced by unilateral condylar neck fracture followed by malpositioned healing was analyzed quantitatively and compared during centric occlusion by combining spiral computed tomography scanning technology with the three-dimensional finite element method.The patient with unilateral condylar neck fracture followed by malpositioned healing had unbalanced stress distribution in the mandible, such as differing nature and value of stress. The stress in the fractured side was higher than in the nonfractured side, in which the condyle was more evident. The maximum principal stress and minimum principal stress of each region in the fractured mandible were much higher than those in a normal mandible. Stress quality also differed.The abnormal stress changes after unilateral condylar neck fracture with malpositioned healing may be related to the effect of condylar shape change on masticatory muscle function and occlusion. TMJ injury not only damaged the condylar structure, but also compromised its biomechanic environment. It is preliminarily thought that unbalanced stress distribution, evidently increased stress, and varied stress character are biomechanic mechanisms of TMD. Condylar dislocated fracture should be treated promptly and properly to maintain the proper prognostic stress distribution.

The exposure of cultured mouse melanoma (B-16) cells, to camptothecin (CAM) after irradiation of the cells with UV-A synergistically enhanced cytotoxicity. CAM treatment prior to irradiation had the same effect. Similar results were also obtained with the combination of gamma-ray irradiation and CAM in this model. Thus, the sensitivity of the cells to UV-A and gamma-ray irradiation was significantly elevated by CAM. These findings have potential clinical utility.

Based on preceding experiment, we further studied the co-regulative effects of PKA-R II and PKC-alpha on expression of oncogenes in human gastric cell line MGC 80-3. The c-myc and c-H-ras expression were suppressed in MGC 80-3 cells during HMBA-induced differentiation. At the same time, PKA-R II showed nuclear translocation from cytoplasm, whereas the expression of PKC-alpha shifted from nucleus to cytoplasm. PKA inhibitor (Sigma) was added to block cAMP-PKA pathway when cell differentiation were induced by HMBA. The PKA-R II was still located in cytoplasm but expression of PKC-alpha translocated again into nucleus. Meanwhile, the c-myc and c-H-ras again expressed. This suggested that the changing regulation of oncogene expression were closely related to signalling from nuclear translocation of kinase subspecies. It thus shows the co-regulation effects of two signal system on oncogenes expression.

We have examined the in vivo targeting potential of the Pleckstrin Homology (PH) domain from human beta I sigma II spectrin using a novel Aequoria victoria green fluorescent protein (GFP) fusion vector constructed from a human codon optimized cDNA. This vector efficiently expresses both GFP and the GFP spectrin fusion protein in COS7 and other cell lines. GFP expressed alone shows only diffuse cytoplasmic staining which is not associated with the plasma membrane. In contrast the GFP-beta I sigma II spectrin PH domain fusion protein localizes under the plasma membrane of transfected COS7 cells in vivo. Fixation of cells transfected with GFP alone in -20 degrees C methanol results in the removal of all specific fluorescence. In contrast cells transfected with the GFP-beta I sigma II spectrin construct and fixed -20 degrees C methanol continue to show strong membrane fluorescence, consistent with a role for the spectrin PH domain in membrane localization in vivo.

Sixty cases with hyperlipoproteinemia (30 cases each group) were observed. The total effective rate of the treated group with Yinchen Wuling Powder was 93.3%. And that of the control group with Gynostemma pentophyllum capsule was 86.7%. The result revealed that significant difference existed between the above-mentioned two groups (P0.01). According to the animal experiment, on both prevention and treatment, this recipe could inhibit the increase of the hyperlipemia model rat's serum TCh, TG, LDL-C and the ratio of LDL-C/HDL-C (P0.01). Besides, this recipe had the effect of marked antioxidation. It could reduce the level of peroxidative lipids, and strengthen the glutathion peroxidase activities.

Quantitative analysis of (l)-ephedrine (l-Ep) and (d)-pseudoephedrine (d-Ps) in plasma by high-performance liquid chromatography (HPLC) is described. The newly developed method is based on a precolumn derivatization with 5-dimethyl-aminonaphthalene-1-sulfonyl chloride (DNSC1) in acetonitrile. The diastereomers formed were separated on a reversed phase column by HPLC with fluorescence detection employing 0.6% phosphate buffer (pH 6.5)-methanol (3:8, v/v) as mobile phase. The detection limit of each Ephedra alkaloid stereoisomer was 0.5 ng at a signal-to-noise ratio of 3:1, the linear response to each stereoisomer being 1-800 ng.ml-1. The plasma level profile of l-Ep and d-Ps in guinea pig was investigated by this method. The determination of l-Ep and d-Ps in plasma of a volunteer after oral administration of Xiao Qinglong Heji was also performed.

Although it has been confirmed that insulin dependent diabetes mellitus (IDDM) is a kind of autoimmune disease, islet cell autoantibodies (ICAs), insulin autoantibodies (IAA) and anti-GAD-glutamic acid decardaxylase antibodies have been found in the sera of patients with IDDM and immunotherapies have been used in some patients with IDDM, yet the manner in which the dysfunctional immunosystem acts on beta cells and causes damage to them remains to be clarified. Recently, possible involvement of the abnormal production of IL-6 in autoimmune symptoms was suggested in patients with cardiac myxomas, Castleman's disease and rheumatoid arthritis. However reports of IL-6 abnormal production in patients with IDDM are rare. In the present study, we examined whether or not an abnormal expression of IL-6 mRNA was present in mononuclear cells (PBMNCs) of the peripheral blood of patients with IDDM. We devised a highly sensitive, specific and semiquantitative protocol, ie, reverse transcription and polymerase chain reaction (RT-PCR). We have used such an assay to measure the relative expression levels of IL-6 mRNA in PBMNCs from 12 early IDDM patients (duration6 mon, 8.20 +/- 3.85 yr), 29 newly-diagnosed NIDDM patients (54.85 +/- 9.12 yr), 23 normal children (8.20 +/- 3.26 yr) and 12 normal adults (31.92 +/- 11.22 yr). In this assay significantly high expression levels of IL-6 mRNA were found in PBMNCs from patients with IDDM (P0.05). The relative levels were 0.91 +/- 0.19; 0.10 +/- 0.06; 0.43 +/- 0.08; 0.10 +/- 0.07, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

By using an inducer of differentiation (HMBA) and blockers of signal pathways, the relationship between two second-messenger pathways (DG-PKC, cAMP-PKA) in inducer-mediated MGc80-3 cell differentiation was studied. Cells were treated with HMBA for 24 h, levels of DG decreased by 64.7%, activity of PKC decreased by 28.7%, while levels of cAMP and rate of it's protein binding increased by 62% and 32.6% respectively (after treated for 48 h, the result was more remarkable). PKA-R II distributed in nuclei. H7 (PKC inhibitor) was used to substitute for HMBA to block the DG-PKC pathway. After treatment for 24 h, the levels of DG and activity of PCK both decreased, while the levels of cAMP increased 1.04 times. On the contrary, PKA inhibitor was added while HMBA was used to induce cell differentiation, cAMP-PKA pathway was blocked, levels of cAMP and rate of it's protein binding decreased. But levels of DG and activity of PKC both increased to the levels of control cells. At this time, PKA-R II distributed only in cytoplasm. These results suggest that harmonious relations of the positive and negative regulation of cAMP-PKA and DG-PKC systems in cells during the proliferation and differentiation of cells. It also showed that positive regulation of PI system may play a leading role in MGc80-3 cells. It let to normal regulation of two signal pathway out of control to remain malignant phenotype of these cells.

The levels of tumor-associated antigens (TAAS) corresponded to monoclonal antibodies WLA-2C4 and CL-3 in sera of 57 lung cancer patients, 100 healthy adults and 50 non-tumor disease patients were assayed with SABC-ELISA of immunobinding inhibition test. The threshold values of WLA-2C4 and CL-3 (RBI) were 12% and 36%, respectively. The positive results of lung carcinomas with at least one of the two TAAS were as follows: squamous cell carcinoma 89%; adenocarcinoma 83%; small cell carcinoma 67% and their mean positive rate was 79%. Whereas the positive rate in healthy adults and non-tumor disease patients was only 6%. These results indicate that using monoclonal antibodies WLA-2C4 and CL-3 simultaneously may be helpful to the serological diagnosis of lung carcinoma.

The serum BGP level was assayed in patients with hyperthyroidism (untreated and remittent cases). The mean serum BGP concentration was 8.7 +/- 2.5 ng/ml in 54 patients with untreated hyperthyroidism, which was significantly higher than normal range (4.8 +/- 1.3 ng/ml P less than 0.01). Serum BGP had a significant positive correlation with the concentration of T3 and T4, while serum AKP had no correlation with circulating thyroid hormone levels. In the patients with hyperthyroidism, serum T3 and T4 decreased significantly after the first month of methimazole treatment, and fluctuated within the normal range since then. Serum BGP did not show significant change during the first six months of treatment, although they were eventually reduced significantly at the end of six months. These results suggest that serum BGP measurement is a valuable marker of bone metabolism alteration during hyperthyroidism.

20 cases of congenital choledochal cyst were operated by modified Lilly's method from 1980 to November 1985 in our Hospital. There were 4 males and 16 females. Their ages ranged from 50 days to 15 years with a mean of 5.1 years. All patients had preoperative ultrasonographic examination, barium meal radiography of the gastrointestinal tract or percutaneous transhepatic cholangiography. All had choledochal cystectomy with retention of the outer layer of the posterior wall of the cyst. We modified Lilly's method by injecting normal saline between the outer and inner layer of the choledochal cyst so that the outer layer could be isolated. Reconstruction of the biliary tract was then performed. 2 patients had choledochoduodenostomy and 18 patients had hepaticojejunostomy (1 end to end and 17 end to side). 19 patients had also had cholecystectomy. The post-operative course of the operation was found to be smooth and safe. Patients were followed up for a period from 6 months to 7 years. 3 patients had cirrhosis of liver, 2 of whom died within 8 months. The mortality rate in our series was 10%. The operative treatment and the problems of biliary reconstruction are discussed.

A 12-year-old boy developed occasional attacks of oculogyric crisis after physical exercises or when tired. Following the initial symptom, progressive Parkinsonian features such as bradykinesia, muscular rigidity, hand tremors in posture, mild dysarthria and disorder of postural reflexes developed. There was no marked diurnal fluctuation o symptoms. Serum ceruloplasmin, copper levels, cranial X-ray CT scan and MRI were normal. Measurement of the plasma levels of L-dopa after single oral administration (300 mg) were normal. The treatment with L-dopa improved the Parkinsonian features excluding the attacks of oculogyric crisis in a few weeks. This case is not identical with juvenile Parkinsonism proposed by Yokochi et al for lack of both crural or truncal dystonia and remarkable response to L-dopa. Oculogyric crisis is known in several patients with severe generalized dystonia, and seldom in patients with Parkinson disease or juvenile Parkinsonism. Oculogyric crisis may be one of focal dystonias confined to extraocular muscles.