From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were analyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse Ig heavy subgroup I (A) and k chain subgroup I, respectively. The VH and VL genes were subcloned into pγ1-Expr and pk-Expr respectively, then transfected into XL2-Blue. The VH-pγ1 and VL-pk were transfected by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques. We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies. These chimeric antibodies are able to kill tumor cells specificallyin vitro.

We prepared an anti-metastatic polypeptide, recombinant fibronectin polypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the function of macrophages. CH50 can significantly augment the production of nitric oxide (NO) by macrophages in a dose-dependent manner. The continuous presence of CH50 had a much stronger effect. In the presence of CH50, the cytotoxicity of macrophages to melanoma B16/F1 cells was significantly enhanced, and a stronger effect was obtained if CH50 was present continuously. CH50 polypeptide and IFN-γ have a synergistic effect on the production of NO by macrophages and the cytotoxicity of macrophages on tumor cells. In thein vivo experiments, CH50 can inhibit the growth of tumor cells, and have a better effect in the presence of IFN-7. Our results suggest that recombinant fibronectin polypeptide CH50 has two functions: one is to inhibit the metastasis of tumor cells, and the other one is to augment the function of macrophages. And this polypeptide will be potentially useful in tumor therapy.

To understand the relationship between the expression of vascular endothelial growth factor (VEGF) and the growth, metastasis of hepatocellular carcinoma (HCC), immunohistochemistry and Northern blot were used to investigate VEGF protein and mRNA in 21 cases of HCC with and without metastasis. VEGF protein was found in 8 of 9 cases with metastasis, whereas only in 4 of 12 cases without metastasis. The positive rate of the former was significantly higher than that in the latter. VEGF mRNA was detectable in both carcinoma and its surrounding liver tissues, but its level in the former was 2–3 times higher than that in the latter. In carcinoma with metastasis, the mRNA level was 5–6 times higher than that without metastasis. It is concluded that VEGF is closely related to the growth of HCC as well as its metastasis and it might be a useful indicator for the metastatic potential of HCC.

The aim of this study was to compare the effects ofd, l-Sotalol andd-Sotalol on the delayed rectifier K⁺ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier K⁺ outward currents were measured in isolated guinea pig single myocytes using the whole-cell configuration of the patch-clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of −40 mV in three experimental protocols [control, isoproterenol (10^-9mollL-10^-6 mollL), and isoproterenol (10^-9mollL-10^-6mollL) plus eitherd, l-Sotalol (10^-4 mollL) ord-Sotalol (10^-4 mollL)]. I_k tail currents were measured upon repolarization to −40 mV. It was found that Ik was significantly amplified in the presence of isoproterenol (10^-9 mollL-10^-6 mollL) plusd-Sotalol. At 10^-8 mollL isoproterenol, I_k was increased by 92. 7%± 17. 1% (P<0. 05) and 54. 3 %±13. 4% afterd-Sotalol addition (P<0. 05). In contrast,d, l-Sotalol completely conteracted the increase of I_k by isoproterenol (<10^-8 mollL), and compared to control, Ik was decreased by 35. 6%± 8. 1 % at 10^-8 mollL isoproterenol plusd, l-Sotalol (P<0. 05). It is concluded that the β-adrenergic blocking property ofd, l-Sotalol but not that ofd-Sotalol maintains the delayed rectifier K⁺ outward current blockade in the presence of isoproterenol in guinea pig myocytes. This might contribute to a superior antiarrhythmic efficacy as compared tod-Sotalol.

The molecular markers of platelet activation, coagulation and fibrinolysis were detected in 60 cases of coronary heart disease (CHD), including 15 cases of stable angina (SA), 21 cases of unstable angina (UA) and 24 cases of acute myocardial infarction (AMI). The results showed that the platelet granule membrane protein 140 (GMP-140) level increased obviously in CHD groups compared with normal control, suggesting that platelet activation existed in CHD. Prothrombin fragment F₁₊₂ and fibrinopeptide A (FPA) were examined to observe the activation of coagulation. No difference was found between SA group and normal controls, while their levels in both UA group and AMI group were significantly higher than in normal control and SA group (bothP<0. 05). D-D dimer and α₂-plasma inhibitor (α₂-PI) were detected to observe fibrinolytic state. The results showed that no difference existed between SA group and normal controls, while both D-D dimer and α₂-PI in UA group and AMI group were significantly elevated than those in SA group and normal controlsP<0. 05).

Fluorescence polarization immunoassay was used to study the chronopharmacokinetics of digoxin in 10 patients with congestive heart failure (CHF) who also served as self-controls. Our results showed that the serum digoxin concentration reached peak value 1 h after taking digoxin at 7:00 a.m., but the serum digoxin concentration reached the peak value 2 h after taking digoxin at 4:00 p.m.. The average serum digoxin concentration area under curve was greater and the best maintainable time of serum concentration within 24 h after taking digoxin at 4 p. m. longer than those at 7:00 a.m.. The heart rates were obviously lower and the cardiac function was significantly improved in 4:00 p.m. group.

The effects of pHGF on the changes of hepatocyte proliferative cycle and liver regenerative capacity after partial hepatectomized rats were observed by flow cytometry (FCM). The results were as follows: 1) S phase fraction (SPF) in group of normal rats (group A) accounted for 9. 89 % and increased gradually within 6 h, following a peak at 12th h or 36th h after operation, but in the group of pHGF-treated rats (group B) the peak appeared at 24th h after operation; 2) Proliferation index (PI) of group A was 19. 6 % before partial hepatectomy, increased to 34. 91 % within 6 h and reached a peaks at 12th or 36th h after operation, and in group B the peak appeared at 48th h after operation. There were significant differences between two groups in SPF and PI (P<0. 01). The weight of liver began to increase 12 h after operation, and almost reached the preoperative weight 5 days after operation. These findings suggest that pHGF can promote the DNA synthesis and segmentation of hepatocyte.

Data obtained in experimental cutaneous melanomas have suggested that the nm23 gene may function as a metastasis suppressor gene. The nm23 level in 8 human cutaneous melanoma cell lines and 2 murine melanoma cell lines were examined. Each melanoma cell line was transplanted subcutaneously into the flank of nude mice, and the metastatic behavior was evaluated by counting lung tumor foci and by determining host survival time. It was found that expression of nm23 mRNA in human melanomas is correlated closely with reduced metastatic behavior in experimental animals and may serve as a sensitive prognostic indicator of malignancy and survival in patients with melanomas.

The model of fulminant hepatic failure induced by acetaminophen was established in dogs to observe the changes of hepatic hemodynamics and plasma histamine levels in portal vein (PV), hepatic vein (HV), abdominal aorta (AA) and inferior vena cava (ICV). The results showed that the portal vein resistance (PVR) was elevated and portal venous blood flow (PVF) was decreased; hepatic artery resistance (HAR) was decreased and the hepatic artery blood flow (HAF), portal venous pressure (PVP), wedged hepatic venous pressure (WHVP) and inferior vena cava pressure (ICVP) had no changes. The histamine of the PV, HV, ICV and AA were all elevated after formation of fulminant hepatic failure. And the increasing wave of the HV was the highest. The increased histamine in HV may be mediated by H, receptor causing the contraction of hepatic venulae, resulting liver sinusoid congestion, increasing PVR and decreasing PVF which exacerbate the liver cell damage. Moreover, the more severe liver damage, the mare histamine was released, and a vicious circle may ensue. Our results also suggest the possibility of using H₁ receptor antagonist to treat the disturbance of liver hemodynamics in severe acute liver damage. The increased histamine in systematic circulation as a vasodilator may lower blood pressure and accelerate heart beats. The increase of plasma histamine may play an important role in the changes of hepatic and systemic hemodynamics in fulminant hepatic failure.

The effect of a new immunosuppressant-I-2190A was tested in a rodent heart allograft model. Grafts were transplanted to recipients heterotopically. There were 5 groups: group 1 received no immunosuppressive agents; group 2 was given CsA (2. 0 mg/kg, i. p.); group 3 was administered I-2190A (0. 1 mg/ kg, i. p.) in carboxymethyl cellulose (CMC); group 4 received injection of I-2190A (0. 5 mglkg, i. p. in CMC)) group 5 received the combination treatment of I-2190A (0. 1 mglkg) and CsA (2. 0 mglkg). Immunosuppressants were discontinued 14 days after operation. No statistically significant difference in grafts median survival time (MST) was found between group 2 (9. 5 days) and group 1 (9 days). The MSTs of grafts in group 3 (22 days,P< 0. 05), group 4 (> 100 days,P<0. 01) and group 5 (> 100 days,P<0. 01) were significantly prolonged compared with control group 1 (9 days). Our results suggest that I-2190A is a potent immunosuppressant able to significantly prolong heart allograft survival in rats after a short time treatment. Low-dose I-2190A could potentiate the effect of sub-therapeutic dose of CsA as well.

Twenty non-insulin dependent diabetics were given nuclide renal dynamic image examination to investigate its significance in the diagnosis of diabetic nephropathy (DNP). The results showed that 1) the glomerular filtration was hyperfunctioned in the diabetics with less than 5 years course, whereas it was impaired in the diabetics with over 5 years course and 2) nuclide renal dynamic image could reveal the glomerular hyperfiltration without β₂-microglobulinuria or microalbuminuria, suggesting that it is superior to the clinical biochemical parameters in the diagnosis, especially in the early identification of DNP.

The protein expression of bcl-2, bax, Fas/Apo-1 in 19 cases of acute myeloid leukemia (AMD were investigated by Western blot and ApAAp techniques. High expression of bcl-2 protein was found in most of the AML cases, and some of the cases expressed Fas/Apo-1 and bax. High expression of bcl-2 protein was associated with a bad clinical prognosis and a poor response after intensive chemotherapy. Bax seemed to coexpress with bcl-2 and so appeared to be a bad prognostic factor instead of a good one. The expression of Fas/Apo-1 was inversely correlated with bcl-2 and seemed to be a good prognostic factor which may reflect the relative homeostasis of apoptotic pathway. It is concluded that apoptosis-induced pathways in AML often exhibit disturbance features. Coregulation of bcl-2, Bax and Fas/Apo-1 genes formed the apoptosis-induced pathway, which is the biological factor affecting response to chemotherapy.

We have shown previously that high-efficient gene transfer can be attained in primary hematopoietic cells using liposome-mediated gene transfer strategy. In order to examine the stability of gene expression mediated by this gene transduction protocol, we observed the expression of marker genein vivo by using bone marrow transplantation (BMT) to engraft lethally irradiated mouse with the genetically modified hematopoietic cells. The results showed that the mouse transplanted with appropriated number of transduced cells remained alive and healthy. The PCR analysis and G418 selection of the spleen colonies and bone marrow cells isolated from lethally irradiated animals 15 days and 30 days after injection of genetically modified bone marrow cells showed that the progeny cells of the transduced hematopoietic stem cells still contained and expressed the transduced genes, suggesting that the hematopoietic system is at least partially re-constructed by the stem cells with marker gene and that the stable expression of foreign genesin vivo can be attained by using this easy and harmless transduction protocol. These findings provide experimental basis for clinician to further investigate the biology of marrow reconstruction and the mechanism of leukemia relapse after BMT.

Vδ2Dδ3 rearrangements of T cell receptor (TCR) gene from cerebral-spinal fluid (CSF) cells was detected for diagnosis and monitoring of central nervous system leukemia (CNSL) in children with acute lymphoblastic leukemia (ALL) using polymerase chain reaction (PCR) technique. 20 patients were studied and in 12 of them the results of PCR and dot hybridization with clonospecific probes were positive, showing the presence of minimal blast cells in CSF. Our study suggested that the PCR method is an effective tool for clinical diagnosis of CNSL and is much more sensitive than routine CSF examination.

In order to explore the effect of biliary tract pressure on Oddi’s sphincter and the mechanism of development of high pressure of biliary tract during acute obstructive and suppurative cholangitis (AOSC), house rabbits were used to establish model of high biliary pressure in acute biliary duct caecus. It was observed that when the pressure of the acute biliary tract was increased to 8 kPa, the electric activity of Oddi’s sphincter was obviously enhanced, the pressure of Oddiœs sphincter increased remarkably (P <0. 05), and even constant spasm appeared with accompanying increase of discharge frequency of the right greater splanchnic nerves (P <0. 05) and progressive decrease of mean arterial pressure. However, when lidocaine of 0. 6 % was used to block the right celiac plexus, no above-mentioned reaction happened when the biliary tract pressure was increased again. The results indicated that the acute biliary tract obstruction might induce the contraction or spasm of Oddi’s sphincter and bring about a vicious cycle. Its mechanism is related to splanchnic nerves reflection and it is one of important factors in the development of AOSC course.

This study was designed to measure the multi-drug resistance gene (MDR1) mRNA content and analyze clinical relationship between MDR1 expression and drug resistance in primary ovarian cancer. Reverse transcription PCR (RT-PCR) was used to measure MDR1 mRNA content in biopsy sample of 31 primary ovarian cancers (experimental group) and 30 gynecological tumors (control group). The level of 95.2% (20l21) MDR1 expression was relatively low, and the detected rate of MDR1 expression was 67.7% (21/31) in experimental group, which was higher than that in control group (40.0%,P<0. 05). The differences of MDR1 expression between the effective group and no effect group after combined chemotherapy was significant (P<0. 05). No significant relationship was found between MDR1 expression and clinical stage or histological classification or grade of differentiation in experimental group. We are led to concluded that primary ovarian cancers have drug-resistance clones which might express MDR1 spontaneously and expression of MDR1 may be used as a prognostic and predictive indicator for clinical response of ovarian cancers to combined chemotherapy.

In order to find an easy and accurate procedure for diagnosis of diffuse goitre in children, we examined 50 patients with diffuse goitre using fine needle aspiration biopsy cytology, thyroid antibody detection, thyroid hormone analysis and ultrasound imaging. In the meantime, 109 healthy children (control) were examined by ultrasound imaging. The results showed that thyroid imaging in health children was a smooth echo pattern with stronger homogenous echogram than surrounding muscle tissues. The patients with diffuse goitre showed an abnormally enlarged thyroid volume. In 22 (84. 6 %) of 26 children with chronic lymphocytic thyroiditis, a varied patch hypoechogenicity was found, of whom 18 (81. 8 %) had positive results of antibody testing. On the contrary, echo-pattern was normal in 17 (70. 8 %) of 24 patients with diffuse thyroid proliferation and only the remainder (7/24, 29. 1%) had abnormal echo-pattern as well as elevated auto-antibody titers, of whom 2 were confirmed as chronic lymphocytic thyroiditis by a repeat fine needle aspiration biopsy 1 year later. By using combined ultrasound imaging and antibody determination, 92% of the cases with chronic lymphocytic thyroiditis could be diagnosed. Our study suggests that ultrasonic imaging is an easy, non-invasive, reproducible and effective procedure in the differential diagnosis of chronic lymphocytic thyroiditis in children.

Experiments were performed on 64 Sprague-Dawley rats under urethane anesthesia. Extracellular recording method was used to investigate the effect of aluminum (AL) microinjected into CA3 on long-term potentiation (LTP) in this area. The relationship between the inhibitory effect of Al and L-arginine-NO pathway was also studied. Microinjection of Al (0.5 mol/L, 1 μl) into CA3 could block the induction of LTP in CA3. Microinjection of Al (0.5 mol/L, 1 μl) into CA3 after LTP was induced could also decrease the amplitude of population spike (PS). The inhibitory effect of Al on LTP in CA3 could be enhanced by preinjection of N^G-nitro-L-arginine (0.3 mol/L, 1 μl). Preinjection of L-arginine (0.3 mol/L, 1 μl) into CA3 could antagonize the inhibitory effect of Al on LTP. These results suggest that Al could block the induction of LTP and decrease the amplitude of PS potentiated in CA3. The effect of Al might be antagonized by L-arginine-NO pathway.