Five hybridomas producing McAbs against human serum IgM-isotype were obtained by fusing the myeloma NS-1 cell and the RALB/C murine spleen cell which had been immunized by serum IgM of patient with B chronic lymphocytic leukemia (B-CLL). These McAbs only reacted with human IgM, not with other immunoglobulins in ELISA and immune double diffusion test. An approximate positive rate of peripheral blood lymphocyte (PBL) was got when these McAbs and the McAbs aginst B cell were tested by indirecl immunofluorescent assay. The positive rate was similar to that obtained by direct immunofluorescent test. Immunoblotting showed that the molecular weight of the antigen to these McAbs was 70–90 Kd, indicating that the antigen was IgM. The practical value of the McAbs against human IgM was also discussed.
In order to search for effective drugs to reduce restenosis incidence after coronary angioplasty, we studied the effects of a Chinese herb, extract of Andrographis Paniculata Nees (APN), and Fish Oil (FO) on atherosclerotic stenosis and restenosis after experimental angioplasty. Preliminary results showed that APN can significantly alleviate atherosclerotic iliac artery stenosis induced by both deendothelialization and high cholesterol diet (control group, stenosis incidence 100 %, stenotic severe degree 60. 53±31. 03 %, of which 30 % arteries (6) are total occlusion; FO group: stenotic incidence and severe degree are 77 % and 53. 00±21.17 %, respectively, and in APN group they are 70 % and 25.39±10. 52 %, respectively,P<0. 01), and follow-up angiography 4 weeks after angioplasty showed that dilated iliac arteries in contrpol group all had severe restenosis, but in APN group no or only mild restenosis occurs, and in FO group restenosis is as severe as stenotic degree prior to angioplasty. These preliminary results suggest that APN and FO can significantly alleviate stenosis induced by deendothelialization and high cholesterol diet and restenosis after angioplasty, while the former has a more marked effect. The above findings lead the authors to conclude that APN may play an important role in preventing restenosis after coronary angioplasty, but FO may be useful in reducing the extent of of restenosis after coronary angioplasty.
results showed tha t , at the baseline, there were no differences in Vs between two groups. During stress , both the Vs and VR of hibernating myocardium were significantly higher than those of necrotic myocardiurn. 95.7 N (29./23) segments with VR 0 were necrotic myocardium while 86 M (6 / 7) segments with V R > 8 0 N were hibernating ones. The distribution of VR was significantly different between two groups ( Z z = 2 1 . a4, P < o . 001). In conclusion, the difference in reaction to low dose dobutamine stress between hibernating and necrotic myocardium were showed by DTI and it is more clinically significant when VR ~ 0 and VR > 8 0 ~ . On the other hand, there are some limitations in this s tudy, the Vs of necrotic myocardium also increased during stress; there were some overlapping parts between two groups when VR was from 0 to 80 M. The following factors might contribute to these limitations: 1) extension of adjacent myocardium; 2) motion of the whole hear t ; 3) placement of the sample volume; 4) difference of the Doppler angle of incidence. To sum up, DTI could show the difference in reaction to low dose dobutamine infusion between hibernating and necrotic myocardium. It has the potential to be used for quantification of regional myocardial function during dobutamine stress and is a valuable addition to the clinical ul t rasound investigations for detecting hibernating myocardium. As a newly developed technique, it still has some limitations. With the accumulation of clinical experience and improvement of the echocardiographic equipment and techniques it will find more application in clinical practice.
ricocele. Fer t i l Ster i l , 1988,49:199 2 . Naz RW, et al. Thymosin al levels in human seminal plasma and fol l icular f luid: implication in germ cell function. Int J Fe r t i l , 1987,32:375 3. Chesnokova VM, et al. Evidence for thymus involvement in development of adrenal and gonadal endocrine functions in mice. Endoerinol J Exp, 1987, 21,133 4 . Ignat" eva EV, et al. Effect of t hy mosin (f ract ion 5) on ~he development of functional ac=ivity of the adrenals and gonads in genetical ly athymic mice. Prol Endocrinol MOSK, 1988,34, 83 5 . Checnokava VM, et al. Effec t of thymosin (fract ion 5) on the endocrine function of the testis in mice. Bull EKSP Biol Med, 1988,I06:493 6 . Raboeh J, Starka L. Hormonal t e s t i eular act iv i ty in men with varieoeele. Ferric Sterol, 197t,22~152 7. Freire FR, Nahoum CR. E~docrine evaluation in infer t i le men with varicocele. Andrologia, 1981,13:395 8 . Clayton RN, Huhtaniemi 1T. Absence of g~nadotropin releasing hormone receptors in human gonadal tissue. Na,ure, 1982,299~56 9 . g .~ ,~ . ~ . s h ~ 5 ~ ? ~ , ~ ~ ~ ~d$~.-~7, i ~ . ~..~fft~/~--~-, 1984,4:60 10. Hal l Nr, et al. Evidence that thymosins and o:her biological response mod i f iers can func':ion as neuractive immunotyansmitters. ]mmu~-..ol, 1985, 1335.'8065 11. Rebar RW, et al. Thymosin s t imulates secretion of huteinizing hormone-releasing factor. Science, 1981, 214:669 12. Pedernera E, ct al. A~hymus factor influences the in Vitro testosterone secretion of leydig cells in the rat. Life Sei, 1986,38z779 (Received May 20,1992)
The effect of Ligustrazine on the hematopoiesis after bone marrow transplantation (BMT) in allogenic BMT mice was investigated. After the typical mice model of allogenic BMT had been established, the mice were randomly divided into three groups: BMT group, Ligustrazine group and normal group. The BMT group was given normal saline (0.2 ml, twice a day) through gastric tube, while the Ligustrazine group was given Ligustrazine through gastric tube (0.2 ml, twice a day). At the 1st, 7th and 14th day after BMT, we observed the peripheral blood cells and bone marrow nuclear cells (BMNC), as well as the expression level of Heparan Sulfate (HS) and stromal cell derived factor-1 (SDF-1) on bone marrow sections by using immunohistochemistry (SABC-AP), the expression of CXCR4 on the BMNC. The results showed that on the 7th and 14th day, the peripheral blood white cells, platelets, BMNC and the expression levels of CXCR4, HS and SDF-1 were significantly higher in Ligustrazine group than in the BMT group (P < 0.05). It was concluded that Ligustrazine could promote hematopoiesis at the early stage of hematopoietic reconstitution after BMT.
To explore the relationship of angiogenesis-related angiopoietin-2 gene and its receptor Tie2 with angiogenesis and the biology, of hepatocellular carcinoma (HOC), angiopoietin-2 gene, Tie2 and CD34 protein expression in 22 resected HOC, 8 cirrhotic and 8 control liver specimens were investigated byin situ hybridization and immunohistochemistry respectively, and the level of angiopoietin-2 and Tie2 expression in HCC were compared in terms of tumor biological parameters. It was found that CD34 was not expressed in control liver, expressed scarcely in cirrhotic liver (17. 8±13.5/HP), but intensively expressed in HCC (86. 3±34. 8/HP,P<0.01). Tie2 receptor was not expressed in controls, expressed at low level in cirrhotic liver (11. 3±8.7/HP), while strongly positive in the microvascular endothelia of HCC (52. 4±16. 7/HP,P<0.01). The level of Tie2 receptor expression in HCC was closely related with tumor diameter, angiogenesis and portal invasion. Angiopoietin-2 gene was not expressed in control liver, expressed mildly in cirrhotic liver (11.2±9. 7/HP), but extensively in tumor zone (36. 4±17. 5/HP), the level of angiopoietin-2 expression was closely related with angiogenesis, portal invasion and histological grading of HCC. It is concluded that angiogenesis is increased in HCC; angiopoietin-2/Tie2 expression in human hepatic carcinoma is closely related with angiogenesis, which are probably involved in the HCC angiogenesis regulation, promoting the development and metastasis of human hepatic cancer.
DNA vaccine plasmids were constructed that encoded two highly-conservative regions of a surface protein, PAc, from the human major cariogenic bacterium, Streptococcus mutans. Antigen expression was evaluated in vitro by immunohistochemical analysis of human endothelial cells following cationic liposome-mediated transient transfection with recombinant plasmid. The results of this study provided a basis for further testing of these recombinant plasmids in primates and for efficacy testing of dental caries DNA vaccines in human volunteers in future.
Presented in this paper were 3 cases of a special kind of polymorphic ventricular tachycardia (VT). The clinical manifestation was recurrent syncope without organic heart disease. The electrocardiogram (ECG) was characterized by normal QT intervals with short-coupled variant of torsade de pointes. The efficacy of treatment with classI,I,I antiarrhythmic drugs was not apparent but verapamil had an excellent therapeutic effect for it. This kind of VT had a high incidence of sudden death, so it was very important for physicians to identify and treat it promptly with long-term verapamil. The mechanism of short-coupled variant of torsade de pointes was unclear. It probably had some relationship with triggered activity or imbalance of autonomic nervous system.
The apoptosis of lens epithelial cells (LECs) induced by ultraviolet and the expression of P53 were investigated. Wistar rats received 100 mW/m2 ultraviolet irradiation (UVR) (λ=280 nm-315 nm) for 15 min. One, 6, 24 h after irradiation the lens capsules were dissected. The percentages of apoptotic cells were evaluated by the TdT-dUTP terminal nick-end labeling (TUNED technique and the expression of P53 was detected by using immunohistochemical assay. The results showed that the percentages of TUNEL-positive nuclei at 24 h after irradiation was significantly higher than in the control group and those 1 h. 6 h after irradiation. The percentages of P53-positive cells at 6 h, 24 h after irradiation were significantly higher than in the control group and those 1 h after irradiation. It was concluded that UVR could induce the apoptosis of lens epithelial cell. The expression of P53 might be responsible for the apoptosis of lens epithelial cells.
In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases of acute leukemia. 139 cases of acute leukemia were enrolled in our hospital from April 1998 to April 2000. Morphological analysis and FCM immunophenotypic tests were conducted on all cases. Our results showed that CD45/SSC gating multiparameter flow cytometry immunophenotyping could reflect the origin of leukemic cells specifically. It is one of the important methods for the diagnosis of ALL, AML, and HAL. CD45/SSC gating multiparameter FCM analysis is a good technique for immunophenotyping. FCM immunophenotypic analysis can help improve the diagnosis and classification of acute leukemia, and extend the use of FCM in clinical practice.
In order to improve the curative effect of onychomycosis, the factors influencing the therapeutic effects were investigated. 545 cases including 245 males and 300 females, who were diagnosed both clinically and mycologically, were treated by Intraconazole with intermittent pulse therapy. The therapeutic effects were judged by the following observations regularly and analyzed from the factors as follows: age; growing speed of nails; accompanied diseases; family history; trauma of nails; infection ways of the pathogens; manifestation of the injury; pathogens; duration of the treatment. The results showed that the recovery rate was higher in younger patients (P<0.01) with a quicker recovery rate (P<0.001), and a lower recurrent rate (P<0.01), as well as in those with quicker growing speed of new-born nail. Also the patients with WSO and DLSO manifestation had a higher recovery rate. The patients with onychomycosis caused by T. rubrum had a higher recovery rate (P< 0.01 to 0.001) no matter whether to prolong the treatment duration. The patients with diabetes mellitus or hyperhidrosis, as well as with positive family history or basic nail diseases such as trauma and paronychia, had a lower recovery rate and the curative effects were not satisfactory. It was concluded that although the single and some DLSO-manifestation nail injury could be cured by internal and external treatments with the help of removing the sick nail and the duration of the treatment could be shortened. The treatment duration should be prolonged in order to increase the curative effects and decrease the recurrence under such conditions as following: old patients above 60 years; patients with low-growing-speed new-borne nails; patients with thumb and big toel injury and ingrowing nail; patients with diabetes mellitus, hyperhidrosis or Renauld’s phenomenon; patients with nail trauma before or during the treatment; patients with PSO or TDO manifestation; patients with onychomycosis caused by Candida or Aspergillus; patients with abnormal new-borne nails of abnormal color, coarse surface or abnormal thickness.
In order to evaluate the value of the ultrasonography in the diagnosis of tumor of the knee and its clinical implication, 67 patients with clinically suspected bone tumor of the knee were examined by ultrasound. The ultrasonographic characteristics of different bone tumors were studied and compared with the results of pathologic characters after operation. Ultrasonography can readily visualize the bony destruction and the pathologic change of the periosteum and the soft tissue related to bone tumor. Fifty-two cases of malignant bone tumors and 15 cases of giant cell tumors were diagnosed by ultrasonography. Pathologically, there were 54 cases of malignant bone tumor and 13 cases of giant cell tumor. It was concluded that ultrasonographic examination might be a useful method for the diagnoses of bone tumor of the knee and play an important role in guiding needle biopsy and electing operative method and approach.
The differences of arrhythmias among distinct left ventricular geometric patterns in the patients with essential hypertension were studied. 179 patients with essential hypertension received 24 h dynamic ECG recording, ambulatory blood pressure monitoring, echocardiography examination, etc. According to the examinations, left ventricular geometric patterns and arrhythmias were identified. The comparison of morbidity of arrhythmias between the left ventricular remodeling group and the normal geometric pattern group was performed. The multiple stepwise regression analysis was carried out to identify the independent determinants of arrhythmias. After these predictors were controlled or adjusted, the severity of arrhythmias among different left ventricular geometric patterns was compared. It was found that the morbidity of atrial arrhythmia, ventricular arrhythmia and complex ventricular arrhythmias in the left ventricular remodeling group was significantly higher than in the normal geometric pattern group respectively. There were many independent factors influencing on arrhythmias in essential hypertension. Of all these factors, some indices of left ventricular anatomic structure, grade of hypertension, left atrial inner dimension, E/A, diastolic blood pressure load value at night and day average heart rate and so on were very important. After the above-mentioned factors were adjusted, the differences of the orders of arrhythmias between partial geometric patterns were reserved, which resulted from the differences of the geometric patterns. Many factors contributed to arrhythmias of essential hypertension, such as grade of hypertension, LVMI, LA, PWT and so on. The severity of arrhythmias was different in different left ventricular geometric patterns.
The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on N1H3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas+ Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8. 5 × 105 colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53. 81 ±6.9 %), and significantly induced the apoptosis of Fas+ Yac-1 cells (56.78±4. 5 %), as both were cocultured for 5 h at 1:1 ratio, whereas it is 7. 08±3. 4 % in control group (P<0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.
Die Primärstabilität von Pedikelschrauben im Knochen hängt in hohem Masse von der Knochendichte ab. Die Augmentation mit Zement ist ein klinisch einsetzbares Verfahren, um die Schrauben-Verankerung im osteoporotischen Knochen zu verbessern. In dieser Studie wurde eine neuartiges resorbierbares Knochenersatzmaterial (α-BSM™) für diese Augmentation benutzt. Biomechanische Testungen wurden in vitro bei 16 Lendwirbelkörpern (L3-L5) aus 6 Individuen (75,2 ±13,7 Jahre) durchgeführt. Vor der biomechanischen Testung wurde bei allen Präparaten die trabekuläre Knochendichte mittels pQCT gemessen und beide Pedikel mit USS-Pedikelschrauben (5, 0 mm X 45 mm) besetzt, von welchen eine mit α-BSM™ augmentiert war. Beim axialen Auszugstest wurden die maximale axiale Auszugskraft (F-max) sowie die Energieaufnahme bestimmt. Der Medianwert der F-max stieg beim Auszugstest durch die Zementierung mit α-BSM™ um 80 % von 370 N (ohne Zement) auf 665 N (mit Zement). Die Energieaufnahme bis zum Erreichen der F-max (E-F-max) und bei Dislokation bis 2,0 mm (E-2 mm) steigerte sich ebenfalls um 83 % und 68 %. Die Unterschiede waren signifikant. (Wilcoxon’s-Test,P<0,01) Die Ausreisskräfte F-max (ohne oder mit Zement) korrelierten eng mit der Knochendichte (r=0,9056 und r=0,9585). Unsere Resultate zeigen, daß eine Augmentation mit dem α-BSM™ die primäre Stabilität von Pedikelschrauben verbessern kann. Der Effekt scheint auf einer Optimierung der Kontaktfläche und einer Aussteifung der schraubennahen Spongiosa zu beruhen. Das Material könnte geeignet sein, die Verankerung von Pedikelschrauben bei osteoporotischen Patieten zu verbessern.
In order to investigate the immunological damage in rat immunized with ATI-receptor peptide. 18 male Wistar rats were divided into two groups: immunized-group (n=12), each rat was immunized with 150 μg AT1-receptor petide coupled to bovine serum albumin, together with Freund’s adjuvant. Control group (n=6), sham-immunized, “immunized liquid” was same as immunized-group except ATI-receptor peptide. Systolic blood pressure (SBP) was measured by using the tail-cuff technique, antibody against ATI-receptor peptide detected by using ELISA method, and left ventricular myocardium and renal cortex sections were observed under light and electron microscopy. There was no significant difference in SBP and light microscopic observation of the tissue sections between the immunized-group and control group. The O.D. value of anti-ATI-receptor peptide antiserum was significantly higher in the immunized-group than in the rats before immunization and control group (P<0.01). Positive rate in the immunized-group was 100%, while 0% in the control group. Ultramicroscopic morphology showed potential myocardial injury, including: increase in number of mitochondria, swelling of many mitochondria with reduction in number or absence of their cristae and cristolysis, disorder of the cardiac myofibrils, and myofibrillar disruption and myocytolysis. And lysosomes were increased in renal tubular epithelia. The ATI-receptor peptide could induce to generate the antibody against ATI-receptor peptide and lead to myocardial and renal damage in rats.
To investigate the effect of calcitonin gene-related peptide (CGRP) on bone resorption mediated by interleukin-1β(IL-1β)in vitro, the osteoclasts isolated from the long bones of newborn SD rats were co-cultured with osteoblasts on ivory slices placed in 24-well plates. 24 h later, conditioned media containing CGRP and/or IL-1β were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL-1β significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL-1β in a dose-dependent manner, It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts.
The mechanism of chemotherapeutic drug-induced apoptosis in leukaemic cells was studied to further investigate whether Fas/FasL system was involved in apoptosis induced by chemotherapeutic drugs and assess their effects when used in combination with soluble FasL (sFasL). The expression of Fas on human leukaemic cell lines K562, HL-60 and U937 treated with daunorubicin (DNR) or cytosine arabinoside (Ara-C) was detected by using flow cytometry. The activities of sFasL, DNR and Ara-C inducing apoptosis of leukaemic cells, in the absence or presence of neutralizing anti-Fas IgG antibody, were detected by using flow cytometry and TUNEL. The results showed that flow cytometric profiles of K562, HL-60 and U937 cells treated with DNR or Ara-C failed to show any significant increase in Fas expression over 18 h (P>0.05). Anti-Fas monoclonal antibody (IgG) could not block the apoptosis in leukaemic cells induced by DNR or Ara-C, but could block the apoptosis induced by sFasL. A role of sFasL in a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs was revealed. It was concluded that chemotherapeutic drug-induced apoptosis in human leukaemic cells (UG37, HL-60) is independent of the Fas/FasL system, but combination of sFasL and drug treatment produces a synergistic cytotoxic effect on human luekaemic cells.
The isolated cardiac myocytes of rats were immediately infected by cosackievirus B3 (CVB3) to investigate the effects of such procedure on the cell cycle, apoptosis and intrazellular ionized calcium (Ca2+ i) of cardiac myocytes. Newborn Balb/c murine cardiac myocytes were cultivated, then infected by CVB3. Intracellular Ca2+ i was measured by flow cytometer. The calcium in the medium for culturing cardiac myocytes was detected by using atom absorb spectrum test. It was found that CVB3 could markedly inhibit the differentiation and proliferation of the infected cardiac myocytes and induce the apoptosis. The intracellular Ca2+ i level in the infected group was significantly higher than in the control group (P<0.01). The calcium concentration in the medium for culturing cardiac myocytes in the infected group was significantly lower than in the control group (P<0.05). It was suggested that the apoptosis and intracellular calcium overload of the CVR3-affected cardiac myocytes are likely to play an important role in the pathogenesis of viral myocarditis.
To investigate the feasibility of using free fetal DNA from maternal plasma as the source of fetal material in non-invasive prenatal diagnosis, SRY gene of free DNA in maternal blood of 65 samples were analyzed by using primer extension preamplication (PEP) and probe microplate hybridization techniques. The results showed that the detection rate of SRY gene in maternal blood from women carrying male fetuses detected by probe microplate hybridization alone and probe microplate hybridization with PEP were 76.09% (35/46) and 95.65% (44/46) respectively, and there was a significant difference between them. The non-detection rate of SRY gene in blood samples from women carrying female fetus was 100% (19/19). It is indicated that probe microplate hybridization was an effective method in detecting trace fetal DNA from maternal plasma and the sensitivity could be substantially improved by combined use of the two techniques. Analysis of fetal DNA in maternal plasma can serve as an alternative for non-invasive prenatal diagnosis.
