The DNA fragments of 150bp length promoter of humanMycobacterium (M.) tuberculosis heat shock protein (hsp) 70 and 650bp length foreign gene, theSchistosoma japonicum glutathione S-transferase (Sj26GST) gene, were obtained by amplification with polymerase chain reaction. And the 150bp DNA sequence Upstream initiation codon ATG ofthe human M. tuberculosis hsp70 promoter that contains the sequenceTTGAG and ATCATA which consensus withE. coli promoter’s -35 and -10 region respectively, as well as ribosome binding site GGAGG at position-12–-8 upstream the ATG were determined by SangerDideoxyribonucleotide-mediated chain-termination method. Then, the human M. tuberculosis hsp70 promoter and Sj26GST cDNA were cloned intoE. coli-mycobacteria shuttle plasmid pBCG-2000 to constructE. coli-Mycobacterium expression shuttle plasmid pBCG-Sj26 that can express Sj26GST gene. The M. smegmatis were electroporated and the positivecolonies were selected by kanamycin. The M. smegmatis containing the vector pBCG-Sj26 can be induced by heating and hydrogen peroxide (H₂O₂) to express GST. The molecular weight of the recombinant GST (rGST) was 26000. The rGST contents that were about 10 percent of the total bacterial protein were analyzed by density scanning after running SDS-PAGE. This study would provide scientific evidences for application of hsp70 promoter in expressing foreign gene in mycobacterium and development of mycobacterium as multiple-valent vectoral vaccine.

The 67KD laminin receptor (LN-R) that binds laminin (LN) is involved in the metastasis cascade. Using immunohistochemical technique,in situ hybridization and reverse transcription polymerase chain reaction (RT-PCR), we studied LN-R protein and RNA levels in 30 cases of human hepatocellular carcinoma (HCC) to further understand its role in the metastasis of HCC. In our 14 cases of HCC with metastasis, its positive rates were 71.4 %, 57. 1 %, 85. 7 % respectively, whereas its positive expression in 16 cases without metastasis were 31.3 %, 18. 8 %, 50.0 % respectively. The significant difference was found between these two groups. The results suggest that the 67KD LN-R expression plays a very important role in the metastasis of HCC.

Monocyte chemoattractant protein-l (MCP-1), a potent chemoattractant, is thought to play an important role in migration of monocytes into atherosclerotic lesions. The present study was designed to investigate the capacity of human peripheral blood monocytes to express MCP-1 and effects of native very low density lipoprotein (VLDL) and oxidized VLDL (OX-VLDL) on the expression. The total RNA was extracted from cultured monocytes, which were exposed to VLDL and OX-VLDL, and the media conditioned by monocytes were collected. MCP-1 mRNA expression was examined by Northern blot analysis. MCP-1 protein in conditioned media was determined by using sandwich ELISA. The results showed that monocytes can express MCP-1 after a 24 h incubation at 37 C, and the expression was markedly increased by a exposure to OX-VLDL, whereas the expression was slightly increased when exposed to VLDL. It suggests that the capacity of monocytes to produce MCP-1 that recruits and activates circulating monocytes may be of considerable importance in atherogenesis, and oxidation of VLDL enhances its potential to promote atherogenesis.

The rearrangement of immunoglobulin heavy chain gene (IgH) and T cell receptor γ gene (TcRγ) was studied in 30 patients with acute lymphoblastic leukemia (ALL) by the polymerase chain reaction (PCR). 19 cases was found to have rearrangement of IgH gene, 12 of TcRγ. Most of IgH rearrangement was characterized by one or two specific bands while some had more than two. Rearrangement of TcRγ gene appeared as one specific band. A slight difference in number, size and lightness of bands was found among the patients. 4 different kinds of rearrangement were observed in the detection of IgH rearrangement in combination with TcRγ gene. The rearranged patterns of IgH and TcRγ gene as well as the clinical significance were discussed.

The effects of the medium conditioned by endothelial cells under hypoxic condition on the phenotype modulation of pulmonary artery smooth muscle cells (PASMCs) were investigated by using cell culture and morphometrical analysis. The results showed that the number of diploid cells and alpha-sm-actin of PASMCs in the hypoxic endothelial cell conditioned medium group (HECCM) was lower and the myofibrilles were even less than that in normal endothelial cell conditioned medium group (NECCM). On the contrary, the volume density of rough endoplasmic reticulum (RER) and mitochondria of PASMCs in the HECCM group was higher than that in the NECCM group, i. e. altering from a contractile type to a synthetic type. However, under direct hypoxic condition for 24 h, the phenotype of cultured PASMCs remained unchanged, i. e. retained contractile type. It suggests that the HECCM can induce phenotype modulation of PASMCs, may be mediated by the pulmonary artery endothelial cells (PAECs) which can secrete some cytokins affecting the PASMCs.

To clarify the role of urokinase plasminogen activator (uPA) in the mechanisms of regulating sperm motility and the ability of fertilizing, we investigated the quantities and activities of uPA in human seminal plasma and on the membrane of spermatozoa. Semens were harvested from 22 infertile patients with asthenospermia and 20 healthy fertile men according to WHO standards. To quantify the membrane-bound uPA in the samples, polyclonal antibodies against human urokinase were employed by means of a sandwich ELISA. The uPA activities in seminal plasma and on the surface of spermatozoa were determined using A-garose-Fibrine-Plate method and the experiment of immunological identification with polyclonal antibodies against urokinase. In lysates of spermatozoa, significantly lower levels of uPA (23. 1±7. 35 mu/10⁶ cells) and uPA activity (5. 13±3. 85 mu/10⁶ cells) were found in patient group as compared to healthy fertile men exhibiting normospermia (29. 89±9. 40 mu/10⁶ cells and 10. 17±6. 18 mu/10⁶ cells). In seminal plasma, uPA activity in patient group (2134 ± 1581. 3 IU/L) was also found significantly lower than that of normal group (3365 ± 1859. 5 IU/ L). Positive correlations were observed between sperm motility and uPA quantities (r=0, 48,P<0. 005), as well as with uPA activities (r = 0. 45,P<0. 005). Thus, it is inferred that membrane associated uPA on human spermatozoa may be related directly to sperm motility and fertility.

Using experimental model of pregnant rats with asymmetrical intrauterine fetal growth retardation induced by passive smoking, the effects of natural herb “Jia Wei Fo Shou San” on erythrocyte membrane calcium, zinc concentration were observed by atomic absorption spectrophotometry. The results showed that the mean fetal birth weight, zinc concentration of erythrocyte membrane were found to be decreased in the model group as compared with the control group. (P<0. 01, F<0. 01). On the other hand, the element of calcium onto erythrocyte membrane were higher in model group than that in control group (F<0. 05). These changes were significantly mild in the treated group and were similar to those of the control group (P>0. 05). Furthermore, our findings indicated that the zinc concentration of erythrocyte membrane seems to be positively correlated with the birth weight (F<0. 01). Calcium composition of red cell membrane showed a significant negative relation to the birth weight (P>0. 05). Our results provided an experimental evidence that normal concentrations of calcium and zinc onto erythrocyte play an important part in fetal growth. One of the mechanisms of “Jia Wei Fo Shou San” in improving fetal growth may have something to do with modulation of erythrocyte calcium, zinc element, thereby protecting bio-functions of erythrocyte and promoting blood circulation.

The activity of Superoxide dismutase (SOD) in renal cortex and erythrocytes, the level of lipid peroxidant (LPO) in plasma, renal function and morphology of leftover kidney were examined in rats. The results showed that Huangdan capsule could slow down the increase of BUN and Scr, promote the SOD activity in renal cortex and erythrocytes, reduce the level of LPO in plasma and delay the renal pathological changes in 5/6 nephrectomized rats. The mechanism of Huangdan capsule’s antiperoxidant effect, its effect on the renal pathological changes and relationship between the two effects were also discussed.

After irradiation with 8 Gy⁶⁰Co γ-ray, mice were immediately given intraperitoneal injection of 200 mg 100% compound blood-activating soup twice a day. On the 3rd and 7th day, the P₅₃ gene expression of bone marrow hematopoietic cells in Chinese drug group was found to be higher than that in normal group, and it was also significantly higher than that in control group. The expression level of GADD₁₅₃ gene which was not expressed in normal group was much lower in Chinese drug group than that in control group. On the 7th day after irradiation, the P53 and GADD153 gene expression levels of splenic mononuclear cells were consistent with those of bone marrow hematopoietic cells both in Chinese drug group and control group. On the 3rd and 7th day, the bone marrow hematopoietic tissue volume in Chinese drug group was higher than that in control group, with no difference found between the two groups. While on the 14th day, the difference became significant (P<0.01). The results showed that commonly used blood-activating and stasis-eliminating drugs may strengthen the viability of hematopoietic cells and promote the rehabilitation of hematopoiesis by inducing wt-P₅₃ expression to block the bone marrow hematopoietic cells in G₁ phase, during which DNA could be repaired.

After irradiated by 8 Gy⁶⁰Co γ-ray, mice were intraperitoneally injected immediately with 0. 2 ml 100 % compound blood-activating soup twice a day for 10 days. The in situ ulnar bone marrow partial pressure of oxygen (PbO₂) was determinedin vivo before, during and after irradiation respectively. The bone marrow sections in the same part were observed. Our results showed that the normal murine ulnar P_bO₂ was 12. 72±1. 05 kPa. During irradiation, the level of P_bO₂ decreased to 10. 78±1. 17 kPa (P<0. 001). And 3 days after irradiation, P_bO₂ decreased to 9. 75±0. 52 kPa, suggesting that the commonly used “blood-activating and stasis-eliminating” Chinese drugs could promote the rehabilitation and proliferation of bone marrow microvessels in the acute radiation injured mice, expand their areas, increase the oxygen supply of bone marrow microenviroment, thereby leading to P_bO₂ much higher increase than that of control group. It is also helpful in the proliferation and rehabilitation of hematopoietic cells.

PAP Immunohistochemical technique was used to detect the expression and distribution of EGF-R in gastric mucosa in portal hypertensive pigs with cirrhosis. Our result showed that the EGF-R content in gastric mucosa of portal hypertensive pigs was significantly lower than that in control group. Microscopically, the portal hypertension (PHT) group imparted “feeble positivity ” or “negative results ” and the control group showed “strong positivity ”. It was suggested that the decrease of EGF-R content in gastric mucosa weakened defense mechanism of gastric mucosa and increase its vulnerability. It is believed that the decreased EGF-R in gastric mucosa is ascribed to the lesions of gastric mucosa during PHT with cirrhosis.

Angiotension-I (A-I) receptor maximal binding capacity (B_max) and dissociation constants (K_d) of different blood vessels in rats with prehepatic portal hypertension were studied by radioligand binding analysis. The results showed that the A-I receptor B_max in the thoracic aorta, superior mesenteric artery and portal vein of portal hypertensive animals (113. 7±19. 4 fmol/mg protein, 206. 9 ±39. 3 fmol/mg protein and 31. 5±9. 2 fmol/mg protein respectively) was all significantly lower than that of controls (146. 8±24. 5 fmol/mg protein, 297. 2±44. 7 fmol/mg protein and 53. 4±12. 1 fmol/mg protein respectively,P<0. 01). The A-I receptor K_d in the superior mesenteric artery was markedly increased in portal hypertensive animals (1. 03±0. 11 nmol/L) compared with that in controls (0. 88±0. 08 nmol/L,P<0. 05). In the thoracic aorta and portal vein, the A-I receptor K_d in portal hypertensive animals was slightly higher than that in controls, but no significant difference was observed between the two groups. The results suggested that the vascular hyporesponsiveness to A-I in portal hypertension was caused partially by a reduction in number and a decrease in affinity of vascular A-I receptors, and these changes might possibly lead to the formation of hyperdynamic circulation.

To evaluate the effect of amino acid cardioplegia on myocardial metabolism and function of ischemic canine heart, canine cardiopulmonary bypass (CPB) model was established and the dog heart was subjected to a 120 min ischemie arrest. Animals were divided into 3 groups, group 1: warm blood cardioplegia induction and terminal perfusion plus 4 C ST. Thomas hospital solution (STS) during ischemia; group 2: warm blood cardioplegia enriched with amino acid (L-asparte and L-glutamate 13 mmol/L each) and STS without amino acid (A. G.) and group 3: both warm blood cardioplegic solution and STS enriched with A. G. The result demonstrated that the cardiac function of animals in group 2 and 3 had a significantly better recovery after ischemic-re-perfusion. By the end of ischemia the content of myocardial ATP in group 3 was distinctly higher than that in group 1 (P<0. 05), with the release of cardiac enzyme being the least. Myocardial ultra-structure almost remained intact before and after ischemia. Our experiment suggests that the cardioplegia arrest with warm blood and cold crystalloid solution enriched with amino acids could diminish the ischemia-re-perfusion injuries of the heart and enhance the effect of myocardial protection.

The purpose of this study was to investigate the effect of non-adrenergic agents on cardiopulmonary resuscitation (CPR) and end-tidal CO₂(ETCO₂) during CPR in a rodent model. Our results suggested that: 1) coronary perfusion pressure (CPP) after drugs infusion was increased significantly by methoxamine, arginine vasoprestn (AVP) and angiotension-I (ANG-II), but not by endothelin-1 (ET-l); 2) ETCO₂ prior to defibrillation was decreased significantly by a pure α₁ adrenergic agents, methoxamine and were increased significantly by non-adrenergic agents, ANG-I and ET-l during rodent CPR: 3) a significant positive correlation between ETCO₂ and CPP was observed in AVP group, suggesting that AVP have little effect on pulmonary circulation; and 4) methoxamine, AVP and ANG-II have similar effect on resuscitability during rodent CPR.

The purpose of the study is to identify a repair procedure for the ulcers or defect of the anterior lateral plantar region. The defects of anterior lateral plantar region after resection of the obstinate ulcer, painful corns and callosity were repaired by the retrograde island flap pedicled with lateral plantar artery. One to 10 years following up revealed that all the 10 flaps in 9 patients survived after operation. Patients walked freely without pain and claudication. The flap did not show any sign of damages of the flap even if the patients walked for a long time. It is concluded that the retrograde island flap pedicled with lateral plantar artery is the best choice in repairing the defects of anterior lateral plantar region.

Eine neue Technik mit Ballonkatheter bei der Gefäßembolisation von einem Aneurysma und einer arterioportalen Fistel wird betrichtet. Es handelte sich um zwei Fälle, bei denen ein Kobrakatheter als Embolisationskatheter und ein Dilatationskatheter als temporärer Okklusionskatheter zur Embolisation angewendet wurden. Durch diese Maßnahme konnten die Aneurysmata und die arterioportale Fistel ohne Spiralreflux verschlossen werden. Die Anwendung von einem Kobrakatheter zusammen mit einem Dilatationskatheter zur Gefäßembolisation ist effektiv. Komplikationen sind relativ selten.

The purpose of this study was to evaluate skeletal and dental effects of bionator headgear combination appliances on patients in development period with Class I, division 1 malocclusion. The comparison of computerized X-ray cephalometric measurements between the 26 treated children and 26 untreated children was made. The results showed that ANB angle was significantly reduced and horizontal mandibular growth development tended to be normal in the treated group. It was suggested that the bionator headgear combination appliance can restrain the maxillary growth early and promote the forward mandibular growth which contribute the functional jaws correction.