To investigate the expression of interleukin-2 (IL-2), metabotropic glutamate receptor subunit 1 (mGluR1) and estrogen receptor (ER) in neurons of the rat central nervous system (CNS) and identify the coexistence possibility of these immune-neuro-endocrine substances in the central neurons, the tri-labeling immunocytochemical technique with different species-specific primary antibodies (goat anti-IL-2 antibody, rabbit anti-mGluR1 antibody and mouse anti-ER anti-body) were used to incubate two serial neighbor sections (one for demonstrating IL-2, another for mGluR1 and ER) of the cerebral cortex, medulla oblongata and spinal cord. There were IL-2-, mGluR1-and ER-immunoreactivity (IR)-positive labeled neurons in the above-mentioned central areas. The IL-2-IR production showed brown color, located in the cytoplasm; In the neighbor serial section, the mGluR1-IR, production showed blue-black color, located on the cell membranes the ER-IR production also showed brown color, located in the cytoplasm and nuclei. There were mGluR1/ER double-labeled cells in the same section, which accounted for about 50 %–60% of the total single and double labeled neurons. It was identified by projection check of serial neighbor sections that had mGluR1/ ER/IL-2 tri-labeled cells, which accounted for about 30% of total mGluR1/ER double-labeled neurons. The results indicate that mGluR1, ER and Il-2 can coexist in the same rat central neurons, therefore, providing morphological basis for the theory about immune-neuro-endocrine network at the cellular level for the first time.
To find a new way for gene therapy against tumors with weak immunogenicity, the effect of mB7-1 costimulation alone, or combined with IL-6, in inducing antitumor immunityin vitro was investigated. It was found that mB7-1 cD-NA transfected B16 cells (B16-mB7-1) induced the expansion of effector lymphocytes and the generation of specific lytic activity more effectively than wild type B16 melanoma cells (B16-wt) or mock-transfected B16 cells (B16-neo) did. (P < 0. 01), IL-6 could effectively stimulate lymphocytes proliferation, but failed to enhance its cytotoxicity, while the combination of mB7-1 and IL-6 increased both lymphocyte proliferative response and T-cell-mediated cytotoxicity more significantly than B7-1 or IL-6 did alone (P<0. 01). It was inferred that the costimulatory molecule B7-1 is required for the activation and proliferation of T lymphocytes; the expression of mB7-1 in tumor cells could increase their immunogenicity and induce effective antitumor immune response, and the combination of B7-1 and IL-6 could induce more effective antitumor immunity, indicating that cooperation of IL-6 and mB7-1 plays a role in T lymphocyte activation.
The model of SD rats ligated at the proximate left anterior descend (LAD) of coronary was used. The number and dissociation constant of β receptor were studied by using receptor autoradiography to observe the changes in β receptor and the effects of Radix Ginseng Rubra on cAMP in experimental ischemic my-ocardium. The result showed that the number of binding site in simple ligation group (Bmax = 0. 279) was obviously higher than that in sham operation group (Bmax = 0. 093) and the dissociation constant of simple ligation group (Kd = 12. 431) was higher than that of sham operation group (kd = l. 319). There was a significant difference between the two groups (P<0. 05). It proved that the number of β receptor was increased and the activity was elevated in myocardial cell membranes after ligation of LAD. The myocardial cAMP level in simple ligation group [(1293. 96±519. 36) × 10-3 nmol/g] was much higher than that in sham operation group [(774. 44 ±210. 55) × 10-3nmol/g]; but the level of cAMP in ligation group after receiving Radix Ginseng Rubra treatment (805. 02 ±362. 48 pm/g) was obviously lower than that in simple ligation group (P<0. 01), which was close to the result of sham operation. The results indicated that Radix Ginseng Rubra could decrease the cAMP level in ischemic myocardium.
The changes of pulmonary arterial and pulmonary venous plasma endothelin (ET) level in oleic acid(OA) —induced lung injury (ALI) rats were observed. After the ALI model of rats was established by intravenous injection of OA, blood samples were taken from a right cardiac catheter inserted via the right external jugular vein into the pulmonary artery to determine the plasma ET levels by using radioimmunoassay before OA injection, and 2 h and 4 h after OA injection. At the same time, arterial oxygen pressure (PaO2) and mean pulmonary arterial pressure (mPAP) were measured to evaluate the role of ET in acute lung injury. The results showed that the plasma ET levels after OA injection were significantly increased in OA group rats as compared with those in the controls, positively correlated with mPAP and negatively with PaO2. The plasma ET level in pulmonary vein was higher than in pulmonary artery, but no significant difference was found. It was concluded that ET, as a local or circulating hormone, might play an important role in pathophysiology of ALI.
The platelet membrane glycoprotein (GP) IIb/IIIa, GPIX were detected by immunogold assay and flow cytometry, respectively, in two cases of Glannzmann’s thrombasthenia (GT). The immunogold assay showed that the GP IIb/IIIa. gold granules in GT were decreased obviously, compared with that in normal controls. Flow cytometry results showed that GP IIb/ IIIa in two cases of GT was 0. 1 % and 0. 5% of normal control, respectively. While GPIX showed no difference between the GT and normal control. The patients’ father showed no bleeding symptoms, whose GP IIb/IIIa accounted for 36. 99% of normal control. The experiment results suggested that flow cytometry is a quick, simple and sensitive method for the diagnosis of GP disorders.
After irradiation by 8. 0 Gy γ-ray, each mouse was stomach-fed by 4 mg ligustrazine injection twice a day. On the 7th day after irradiation, CD49d expression in ligustrazine-treated group was significantly higher than that in control group (P<0. 01), and showed no difference from that in normal group (P> 0. 05). On the 14th day after irradiation. CD49d expression was increased in control group, but decreased significantly in ligustrazine-treated group (P<0. 01). The expression of Cyclin D2 in spleen mononuclear cells (MNC) in ligustrazinetreated group was significantly higher than that in control group, but the ratio of G0+G1 phase cells was significantly lower in ligustrazine-treated group (P<0. 01). This finding indicated that ligustrazine could increase the expression of adherent molecule on bone marrow hematopoietic cells and Cyclin D2 in spleen MNC, thereby promoting the growth of hematopoietic cells.
The aggregation and ATP release of placelet of normal subjects were measured by platelet lumi-aggregometer. It was found that the aggregation curve induced by SJAMP at the concentration of 100 mg/L was a typical second phase aggregation. There existed a certain lag between platelet aggregation and secretion. The secretion actually began slightly after the second phase of aggregation, suggesting that the second phase aggregation induced by SJAMP is not dependent upon the release of contents of dense granule alone. If platelets were incubated with cyclo-oxygenase inhibitor, the second phase aggregation was inhibited and no ATP was released. The results indicated that the aggregation and release reaction induced by SJAMP were dependent upon the generation of prostaglandin endoperoxides and TXA2 in normal subjects. The amount of ATP release was 0. 69 ±0. 22 nmol/108 platelets as stimulated with SJAMP (100 mg/L). But the amount of ATP release were 1. 60±0. 25 and 1. 37±0. 15 nmol/108 platelets when platelets were stimulated with ADP (5 ώmol/L) and collagen (5 mg/L). The amount of ATP release induced by SJAMP was significantly lower than that of ADP and collagen. These findings indicated that SJAMP was a weaker agonist than ADP in terms of platelets release reaction.
The effects of serum of patients with Kawasaki disease on the platelet derived growth factor-B (PDGF-B) chain protein expression in monocytes were studied by immunocytochemical method and the effects of the serum on the endothelial cellular (EC) apoptosis were observed by flow cytometric technique. It was found that the serum of patients with Kawasaki disease induced significantly the expression of PDGF-B chain protein. Likewise, EC apoptosis was increased significantly in the experimental group as compared with the control group (P<0. 01). The results suggest that PDGF in monocytes increase and EC apoptosis play an important role in the development of coronary artery complication in Kawasaki disease.
Experimental mice were immunized with cardiac myosin and complete Freund’s adjuvant (CFA) on days 0, 7 and 30 and control mice were immunized with CFA. Sera and myocardium samples were harvested on days 15, 21 and 120 after the first immunization. Pathological findings demonstrated that there were necrosis and inflammatory infiltration in acute stage and fibrosis mainly in chronic stage of experimental group. Anti-myosin antibodies were also found in sera of experimental mice, but not in control group. Our study showed that cardiac myosin served as an autoantigen to provoke autoimmunity and lead to the transformation of VMC into DCM.
Hepatitis G virus (HCV) isolates from 12 patients with chronic hepatitis C underwent the sequence analysis. Among the 12 patients, 9 obtained a complete response (CR), 2 partial response (PR) and 1 non—response (NR) after the treatment with IFN-α. The results showed that only single amino acide (aa) substitution in iIFN sensitivity determining region (ISDR) (aa 2213 and aa 2218) in 6 cases with CR was observed, while 3 cases of CR and all PR and NR cases had no aa mutation of ISDR. It is suggested that the ISDR in HCV NS5A was yet not identified by our patients.
To investigate the intracellular mechanism that interleukin-1β (IL-1β) facilitates epileptic seizure and neuronal damage, the effect of IL-1β alone or IL-1β plus glutamate (Glu) on the intracellular free calcium ([Ca2+]i) of single cultured hippocampal neuron was examined by using EPC-9 light-electricity measurement system. The results showed that IL-1β of different concentrations (5 × 103 U/L, 10 ×103 U/L, 20×103 U/L, 30×103 U/L, 50×103 U/L, 100×103 U/L) failed to affect the neuronal [Ca2+]i, but IL-1β could facilitate the augmentation of neuronal [Ca2+]i induced by Glu in a dose-dependent pattern. MK-801 inhibited the effect of Glu on [Ca2+]i, and also inhibited the effect of IL-1β on [Ca2+]i induced by Glu, while verapamil did not influence the effect of Glu or IL-1β. It is concluded that IL-1β, as a neuromodulator, can facilitate the activation of NMDA receptor by Glu, induce the increase of intracellular calcium, which enhances the excitement of neuron.
To elucidate the pattern of paclitaxel-induced apoptosis in human ovarian cancer cell line COC1 and the role of paclitaxel in chemotherapy of ovarian cancer, apoptosis was investigatedin vitro by applying cytohistochemical techniques, DNA gel electrophoresis and flow cytometry. COC1 cells manifested typical apoptotic morphologic features after exposure to paclitaxel. The rate of apoptosis was enhanced within the test concentration range in a concentration-dependent pattern. At low paclitaxel concentration, the rate of apoptosis were low and the levels of mitotic arrest were high. Whereas at higher paclitaxel concentration, the rate of apoptosis were higher and the levels of mitotic arrest were relatively lower. It is concluded that the antitumor effect of paclitaxel was correlated with druginduced apoptosis. Apoptosis induced by paclitaxel was concentration-dependent and was not significantly correlated with the mitotic arrest.
To study the effect ofHelicobacter Pylori (Hp) on the process of gastric carcinogenesis, 35 cases of chronic gastritis, 20 cases of gastric adenocarcinoma were studied by use of transmission electron microscopy, immunohistochemical and molecular biological technique. The results showed that 24 of 35 cases of chronic gastritis were positive for Hp, 11/20 cases of gastric adenocarcinoma wereHp positive. PCNA positive cell labeling index (LI) inHp-associated chronic gastritis (LI= 20. 6±4. 7) Was higher than that inHp negative chronic gastritis (LI = 11.3±5. 2) (P<0. 05). HSP70 expression of gastric adenocarcinoma tissues inHp-infected patients were lower than that of non-Hp-infected gastric cancer. p53 gene mutation was found in gastric adenocarcinoma with positiveHp. It was suggested thatHp may enhance gastric cell proliferation, decrease the expression of HSP70 which induces p53 mutation.
To study the effects of tricalcium phosphate (TCP) ceramics on osteoblasts, the rat osteoblasts were cultured with the TCP ceramicsin vitro. Scanning electron microscopy and the colorimetric methyl-thiazol-tetrazolium assay showed that the osteoblasts could adhere well to the surface of the ceramics and the culture dish, and the proliferation of the cells was not inhibited. The results demonstrated that TCP ceramics possessed an excellent cytocompatibility with the osteoblasts, and had some promoting effects on proliferation of osteoblasts.
The hemodynamic changes of portal vein before and after intra-arterial infusion of ethanol iodized oil emulsion (EIOE) were studied in 4 healthy dogs. Segmental transcatheter arterial embolization (TAE) using EIOE was performed in 55 patients with hepatocellular carcinoma (HCC). Twenty patients subsequently underwent surgery. It was concluded that segmental TAE might lead to double embolization of both hepatic arteries and portal veins. It is a more effective therapeutic method for HCC localized in one or a few segments, as compared with conventional TAE.
To observe the clinical effects and the mechanism of Chanlibao (CLB, a preparation of Chinese herbal medicine) in accelerating second stage of labor, primiparae were divided into 3 groups at random. CLB or oxytocin (OTC) was given to the CLB group (n = 80) and the OTC group (n = 52) respectively. The third group served as controls (n = 29). The control group consisted of women experiencing natural labor and to whom no drug was given. The time of second stage of labor and prognosis of mother and newborn of different groups were observed and compared. And intrauterine pressure and fetal heart rate were monitored by means of electronic monitoring. Isolated uterine muscular tissue was used to observe the reactivity to CLB. The results showed that the time of second stage of labor and postpartum hemorrhage in the CLB group were less than those in the control group and the average intrauterine pressure in the former was higher than that in latter, so was the contraction strength of isolated uterine muscle, but with no difference as compared with the OTC group. No side effect of CLB was found. It is concluded that CLB could obviously strengthen uterine contraction and accelerate second stage of labor. Moreover, it is inexpensive, convenient and free of side effect. It can be used as a new, safe and effective alternative for improving prognosis of mother and newborn, especially those not indicated for oxytocin or profuse infusion.
Polymerase chain reaction (PCR) technique combined with direct detection by silver staining on denaturing DNA sequencing gel was used to analyze the (CGG)n repeats within the FMR1 gene on 169 suspected patients with mental retardation and 33 kindreds of 6 fragile X families. The results showed that: (1) No PCR products were detected in 3 males in the suspected group. (2) In the fragile X family studies, the 5 male probands failed to show any PCR products. (3) Diplex PCR with the primers flanking the FRAXE locus was used to serve as an internal control for the 8 above-mentioned males and only normal products of the FRAXE locus were detected, indicating that the possibility of false negative results of the FRAXA locus could be eliminated. These findings suggested that analysis of (CGG)n repeat within the FMR1 gene by PCR technique could efficiently detect premutation carriers and that negative PCR products in mentally retarded males might highly imply the diagnosis of fragile X syndrome after the false negative results have been excluded by diplex PCR. This PCR assay is suitable for the screening and diagnosis of fragile X syndrome in a large number of populations due to its rapidity, simplicity, stability and reliability.
In order to inquire into standard for morphologic classification of choroidal invasion of retinoblastoma and to study the tumor clinic—pathologic further under the integrated scale, the changes of retinal pigment epithelium, Bruch’s membrane and choroid on the histopathological sectins of 297 cases of primary enucleated eyes of retinoblastoma were retrospectively observed under the light microscopy. The choroidal invasion was morphologically divided into four stages: Stage 1, only retinal pigment epithelium involved, the Bruch’s membrane was intact; Stage 2, the Bruch’s membrane was destroyed and the choroidal capillaris was not infiltrated; Stage 3, choroial capillary and middle blood vessel layer in small limits were infiltrated; Stage 4, invasion involved in choroid in great limits and involvement of sciera existed simultaneously. This classification (also called pigment epithel — choroid stage, PEC — stage) reflected both infiltrated procedure of tumor cells and preventive mechanism in eye and morphological criterion half—quantitatively. It can be used as a united standard to compare the infiltration degree of retinoblastoma among different individuals and different studies.
The expression of p16 gene in normal salivary acini, benign tumors and carcinomas were microscopically and quantitatively observed by using immunohistochemical method LSAB and CMIASWIN methods. The expression of p16 gene were found in all 3 groups. The positive unit (PU) was higher in tumor group and cancer group than that in normal group (P<0. 01). Furthermore, the PU of p16 was stronger in cytoplasm than in nucleus. Malignant tumors and acini surrounding the tumor revealed strong positives and weak positives respectively. The PU of pl6 gene was higher in deep lobe of recurrent parotid neoplasm with incomplete capsule than that in shallow lobe of primary parotid neoplasm with complete capsule. The findings suggests that p16 gene plays an equally important role in the salivary gland tumors and tumors in other part of the body.
Nine patients with neuroblastoma were surgically treated and postoperatively followed up. Of the 9 cases, 8 survived well while the other one died of extensive metastasis within cranium 2 years after the operation. One patient had stayed alive for over 12 years.