The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eye-balls were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages. By using ELISA kit, the levels of interleukin-2 (IL-2) and interferon-γ (IFN-γ) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 10⁶ CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference (P>0.05), but both were significantly increased as compared with that in the control group (P<0.05); The contents of NO in the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than in the control group (P<0.001) and rBCG-Sj26GST vaccine group (P<0.01): The levels of serum IL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P<0.001), vector group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05); The contents of serum IFN-γ in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05). The contents of IFN-γ in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P<0.001) but were significantly increased as compared with that in the control group (P<0.01). It was indicated that both vaccines could enhance the immune response of the mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine.

To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method. An animal model of smoking was used for this study. The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke (P<0.01) and then restored to normal level. This fluctuation was associated exclusively with the alteration in number of apoptotic cells (P<0.01). There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups (P>0.05). All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse.

In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1α (MIP-1α), the expression of MIP-1α protein in the cells was detected by cell enzyme-linked immunosorbent assay (ELISA) and that of MIP-1α mRNA was determined by cell in situ hybridization and nuclease S1 protection assay after the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activity of MIP-1α was tested by micropore filter method using modified Boyden chambers. Cell ELISA showed that the expression of MIP-1α protein in endothelial cells exposed to 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the control cells, which was statistically significant by analysis of variance. In situ hybridization revealed that the mRNA expression of ECs treated with 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.3-fold, 3.0-fold and 1.7-fold as much as that in the control group, which had statistical significance (F=188. 93,P<0.01). The mRNA expression in 5 μmol/L dimide treated ECs, measured by nuclease S1 protection assay, was 3.4-fold as much as that in the control group (t=8.70,P<0.05). Chemotactic response (99.50±4.31 μm) to the culture medium conditioned by 5 μmol/L diamide treated ECs, which was stronger than that (66.47±3.25 μm) conditioned by the ECs (F=404.31,P<0.05), was significantly decreased (F=192.25,P<0.05) after adding MIP-1α antibody. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce high level of MIP-1α, and might play an important role in atherogenesis by promoting the migration of peripheral blood monocytes into arterial intima.

To study the influence of recombinant endostatin on angiogenesis and tumor growth of mice H22 hepatoma, tumor models were constructed by injecting H22 hepatoma cells into the leg muscle of mice. Recombinant endostatin was produced by gene engineering in E. coli. The recombinant protein was injected subcutaneously to treat transplanted hepatoma faraway. The weight of tumors was measured, and the changes of necrosis of tumor cells and vessel density were observed by immunohistochemistry. The results suggested that the growth of hepatoma models transplanted in the muscle of legs was suppressed by recombinant endostatin. The density of vacularity was decreased, but the necrosis of tumor cells increased. The inhibitory effect of recombinant endostatin on angiogenesis and tumor growth of hepatoma was not affected after chemotherapy.

To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-κB were labled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-κB consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-κB in PMA group was more than that in PMA+PDTC group. It is suggested that detection of NF-κB by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.

To explore the potential of lipoteichoic acid (LTA) induced cardioprotection against ischemia-reperfusion (I/R) injury in isolated rat hearts and whether endogenous nitric oxide (NO) participates in the protection, the rats were pretreated with LTA (1 mg/kg, i. p.) 24 h before the experiment, and the isolated hearts were subjected to 30 min no-flow normothermic global ischemia and 60 min reperfusion after a 20-min stabilization period by the langendorff method. Cardiac functions were evaluated at the end of stabilization, and at 30 min, 60 min of reperfusion. The amounts of MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH) and total NO oxidation products in the coronary effluent were measured spectrophotometrically at the end of reperfusion. It was revealed that pretreatment with LTA could significantly improve the recovery of cardiac function, reduce the release of CK-MB and LDH, and increase the concentrations of NO in coronary effluent. The protective effects were abrogated by pretreatment of the rats with L-NAME. It was concluded that LTA could induce the delayed cardioprotection against I/R injury, and endogenous NO may be involved in the mechanisms.

The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt, a fragment of the cholecystokinin C-terminal octapeptide CCK-8, was reported. This fragment was synthesized by coupling Phac-Met-OEt with Gly-OMe, Trp-OMe and Met-OEf successively. These three steps were catalyzed by α-chymotrpsin, Papain and α-chymotrpsin respectively. The results of FAB-MS showed that all the products had the correct molecular mass.

To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared. The clinical manifestations, ultrastructure and immunophenotype were analyzed. The results showed that hairy cell leukemia (HCL), splenic lymphoma with villous lymphocyte (SLVL) and hairy cell leukemia-variant (HCL-V) had some common characters including splenomegaly, peripheral blood and bone marrow infiltration by villous lymphocyte and B lymphocyte immunophenotype; but these three disorders had specific features respectively. It was concluded that overall analysis of clinical and laboratory features might be contributive to the differential diagnosis of these three disorders.

To investigate the distribution of variant genotypes of Fc gamma receptor III a (FcγRIII a) in healthy Chinese population of Zhengzhou city, genomic DNA was extracted from peripheral blood of healthy donators. The genotypes of FcγRIII a-158 were determined by nested polymerase chain reaction (PCR) in 137 healthy people in Zhengzhou city. The results showed that frequencies of variant genotypes FF, VV and VF were 42.3%, 48.9% and 8.8% respectively. The distribution of FcγRIII a-158 in healthy Chinese population of Zhengzhou city was polymorphic and different from that of African Americans (AA) and Caucasian Americans (CA).

An extended 5-generation family has been investigated in which 32 of the 111 family members were diagnosed as having retinitis pigmentosa (RP). The proband was a 58-year old male in whom night-blindness was first observed in early childhood, with almost loss of vision by 52 years of age. The symptoms observed in other family members included night-blindness, impaired vision and visual field loss. Dementia, digital abnormalities, deaf-mutism and mental retardation were variously diagnosed in a number of individuals with RP. The affected and unaffected family members were tested for mutations in a range of candidate genes. The 8 exons of three candidate genes have been analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing techniques. A novel mutation was identified in the rhodopsin gene at codon 52 of exon 1 (TTC-TAC) that resulted in a substitution of Phe to Tyr.

In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, Top II_α, bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerse chain reaction (semi-RT-PCR). It was found that the cyclin A and Top II_α mRNA expression levels in drug resistant group were significantly lower than in sensitive group (P<0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group (P<0.01). cyclin A and Top II_α gene expression levels were closely correlated (r₃=+0.794,P=0.000,n=64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 (r₃=−0.337,P=0.029). The 10 AL patients with positive lower expression of both cyclin A and Top II_α were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and Top II_α gene expression would predict drug resistance in AL patients.

In order to explore the molecular mechanisms of sodium butyrate and trichostain A on K562 cell proliferation/differentiation, K562 cells were grown in the absence or presence of sodium butyrate or trichostatin A. The percentage of viable cells was determined by trypan blue exclusion. Differentiation was determined by nitro-blue tetrazolium (NBT) reduction and cell surface adhesion molecules analyzed by FACS. Cell cycle distribution was studied after DNA staining by propidium iodide. Cell cycle regulatory proteins were detected by Western blot and reverse transcription-polymerase chain reaction. The results showed that sodium butyrate blocked cells mainly at the G0/G1 phase of the cell cycle, whereas trichostatin A arrested the cells at G2 phase. Sodium butyrate could down-regulate the mRNA expression of cyclin D1, but not affect its protein expression, down-regulate the protein expression of cyclin D3, but not affect its mRNA expression. Trichostatin A showed similar effects on cyclin D1 and D3 as sodium butyrate. Both sodium butyrate and trichostatin A could stimulate p21 expression of K562 cells at mRNA and protein levels. It may be concluded that sodium butyrate and trichostatin A could promote the proliferation/differentiation of the K562 cells, which might be contributed to the induced expression of cyclin D3 and p21 proteins.

To study the effect of glutamate on the intracellular calcium signal of pure cultured rat astrocytes and the role of NMDA and AMPA receptors in the procedure, the change of calcium signal was investigated by monitoring the fluctuation of intracellular Ca²⁺ concentration ([Ca²⁺]_i) on the basis of Fura-2 single cell fluorescent ratio (F345/F380). The changes in the effect of glutamate on the intracellular calcium signal were observed after blockage of NMDA and (or) AMPA receptors. It was found that L-glutamate could induce an increased [Ca²⁺]_i in most of the cells in concentration-and time-dependent manner. D-(-)-2-amino-5-phosphonopentanoic acid (D-AP-5, a selective antagonist of the NMDA receptor) and 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX, a selective antagonist of the AMPA receptor) could abolish the effects of NMDA and AMPA respectively. The treatment of D-AP-5 and CNQX simultaneously or respectively could attenuate the effect of L-glutamate at varying degrees. All these indicated that glutamate could modulate intracellular Ca²⁺ of pure cultured rat astrocytes through different pathways. The activation of NMDA and AMPA receptors took part in the complex mechanisms.

To study behavioral character and changes of neuronal activity in the basal ganglia of rat model of levodopa-induced dyskinesia, unilateral 6-hydroxydopamine lesioned rat model of Parkinson disease (PD) was treated with levodopa/benserazide twice daily for 4 weeks and the behavior observed on the 1st, 3rd, 4th, 7th, 9th, 10th, 14th, 21st and 28th day. The animals were sacrificed and immunohistochemical technique was used to measure the changes of Fos expression in the caudate putamen (CPU), globus pallidus (GP) and sensorimotor area of cerebral cortex 2 h after the last treatment. The results showed that pulsatile treatment with a subthreshold dose of levodopa gradually induced abnormal involuntary movement (AIM), including stereotypy (limb dyskinesia, axial dystonia and masticatory dyskinesia) towards the side contralateral to the dopamine-denervated striatum and increased contraversive rotation. The motor pattern of each subtype was highly stereotypic across individual rats, and the proportion of each subtype was not consistent among individual rats. Fos positive nuclei in the CPU and GP were increased by levodopa acute administration, and more remarkably in the CPU, but not in the cerebral cortex. After repeated levodopa treatment, Fos positive nuclei were reduced remarkably in the CPU, but were increased in the GP and cerebral cortex. It was concluded that the neural mechanisms underlying levodopa induced AIM in rat model of PD was very similar to those seen in levodopa-induced dyskinesia (LID) in PD patients and MPTP-lesioned monkeys, and increased striatopallidal neuronal activity might be involved in occurrence of LID.

In order to study the relationship between serum C-reactive protein (CRP) levels, leukocyte count and carotid plaque in patients with ischemic stroke, carotid duplex examination was performed by high-definition imaging (HDI) 5000 triplex system. Serum CRP was measured by nephelometry within 72 h after index ischemic stroke. A lesion was considered a plaque in the presence of a maximum intimal-medial wall thickness (IMT) 1.2 mm. Results of carotid ultrasonography were divided into two groups: M1, normal (IMT<1.2 mm) and M2, abnormal (IMT≥1.2 mm). The results showed that the mean age of M2 was significantly older than that of M1 (69.7±10.4 versus 62.5±9.6,P=0.001). The patients with hypertension and diabetes mellitus (78%, 35% respectively) in M2 were significantly more than those (52%, 18% respectively) in M1 (P<0.01,P<0.05). There were 32 (65%) patients with elevated CRP levels in M2, but 33 (46%) patients with elevated CRP levels in M1, with the difference being significant between the two groups (P<0.05). The levels of serum glucose and leukocyte count (8.1±5.5, 10.3±4.0, respectively) in abnormal CRP group were significantly higher than that of normal CRP group (6.4±2.8, 8.7±3.4) (P<0.05,P<0.05); elevated CRP levels was found in 42 (62%), patients with territory infarction and 23 (43%) patients with lacunar infarction respectively, with the difference being significant between these two groups (P<0.05). It was concluded that the elevation of CRP levels was an significant clinical index for carotid plaque in patients with acute cerebral infarction.

To explore the effect of technetium-99 conjugated with methylene diphosphonate (⁹⁹Tc-MDP) on IgM-RF, IgG-RF and IgA-RF (RFs) 47 cases were selected for study, including 33 patients with rheumatoid arthritis (RA) and 15 patients with joint pain/arthritis. After⁹⁹Tc-MDP for drips model being given to the patients by intravenous drip 0.2 g daily for 5 days, the injection A and B models of⁹⁹Tc-MDP were used to the patients by intravenous injection one set daily for 10 days, that was one course of treatment. The next course started after 10 days. Each case used it from 2 to 4 courses of treatment. The RFs in serum were determined by the method of enzymelinked immunoabsorption assay (ELISA) before and after 2 and 4 courses of treatment. In the patients with RA, the concentrations of IgM-RF were 296.2±108.4, IU/ml, 189.5±92.3 IU/ml and 107.8±72.5 IU/ml; the concentrations of IgG-RF were 325.6±126.2 IU/ml, 209.7±98.2 IU/ml and 160.2±80.8 IU/ml; the concentrations of IgA-RF were 330.4±136.3 IU/ml, 210.7±89.2 IU/ml and 148.8±72.2 IU/ml before and after 2 and 4 courses of treatment, respectively. The concentrations of the above RFs were significantly lower after 2 and 4 courses than those before treatment (P<0.05 andP<0.01). There was no significant difference in RFs concentrations in the patients with joint pain/arthritis before and after use of⁹⁹Tc-MDP. In the patients with positive RFs before treatment, the RFs concentrations were decreased significantly after 2 and 4 courses of treatment (P<0.05 andP<0.01). There was no obvious change of RFs concentrations in the patients with negative RFs after treatment of⁹⁹Tc-MDP. It was concluded that⁹⁹Tc-MDP could obviously reduce the abnormally high concentrations of RFs, but not influence the normal RFs, which indicated that⁹⁹Tc-MDP has an important effect on controlling the activities of RA.

To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n=10) or MMF (20 mg. kg⁻¹, d⁻¹, by daily gastric gavage,n=10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), α-smooth muscle actin (α-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.

The expression, activity and clinical implication of heme oxygenase-1 (HO-1) in the chronic renal insufficiency (CRI) rat kidney and its mechanism were investigated. The 5/6 nephrectomized rats were assigned to sham operation group, CRI group and Hemin group. At the 8th week after second operation, blood pressure, urinary protein, serum creatinine (Scr) and BUN were measured. Renal pathologic changes were observed. The activity of HO and contents of erythropoietin (EPO) in serum and renal tissue were determined. Immunohistochemistry was used to detect the expression and distribution of HO-1 in the CRI rat kidney. As compared with CRI group, the urinary protein, blood pressure, Scr and BUN in Hemin group were reduced significantly (P<0.05). The glomerular mesangial proliferation, inflammatory cellular infiltration of renal interstitium and interstitial fibrosis were ameliorated significantly. Immunohistochemistry and measurement of HO-1 activity revealed that the expression and activity of HO-1 was decreased in renal tissues and increased in serum in CRI group as compared with normal rats. HO-1 distributed mainly in tubular epithelial cells. The EPO contents in Hemin group were significantly higher than in CRI group. Through up-regulating the EPO level in serum and renal tissues, HO-1 retards the progression of CRI.

The changes of adrenomedullin (ADM), endothelin-1 (ET-1) and nitric oxide (NO) levels before and after operation in congenital heart disease (CHD) associated with pulmonary hypertension (PH) were observed in order to investigate their role in CHD with PH and their clinical significance. The CHD patients were divided into 3 groups according to pulmonary artery systolic pressure (PASP); Non-PH group: PASP≤30 mmHg (n=11); mild-PH group; PASP 31–49 mmHg (n=10); moderate or severe-PH group: PASP≥50 mmHg (n=12). The control group consisted of 15 health children. Plasma ADM, ET-1 and NO levels were determined by radioimmunoassay and colorimetry methods. The correlation between ADM and ET-1, NO, PASP was analyzed. The changes in plasma ADM, ET-1 and plasma NO on the 7th day after operation among the groups were compared. The results showed that plasma ADM levels in non-PH group were significantly higher than that in control group (P<0.05), but there was no significant difference in ET-1 and NO levels between the two groups (P>0.05). ADM and ET-1 levels in mild-PH group were significantly elevated as compared with those in non-PH group (bothP<0.05), but NO levels were decreased (P<0.05). ADM and ET-1 levels in moderate or severe-PH groups were increased as compared with those in mild-PH group (bothP<0.01), but NO level significantly declined (P<0.05). On the 7th day, after operation, plasma ADM and ET-1 levels in PH group were significantly decreased (P<0.05,P<0.01) as compared with those before operation, but there was no significant difference in NO levels (P>0.05). But NO levels in non-PH group were significantly increased (P<0.05). Plasma ADM, levels in CHD were positively correlated with PASP and ET-1 (r=0.77,P<0.01;r=0.82,P<0.01), negatively correlated with NO (r=−0.56,P<0.05). It was concluded that during the progression of PH in the cases of CHD, plasma ADM, ET-1 and NO might play an important role in the development of PH. The increased ADM may represent a compensatory mechanism. It can interact with NO and ET-1 to regulate pulmonary circulation in the pathophysiology of PH with CHD. ADM may be involved in the defence mechanism against further increase of pulmonary arterial pressure. ADM could be used as a reliable indicator of the severity of CHD associated PH.

In order to assess the diagnostic value of invasive electrophysiologic study (EPS) in the patients with unexplained syncope, the electrophysiologic findings of 268 patients with unexplained syncope despite a complete clinical evaluation were analyzed. Results showed positive EPS finding was 38% in total patients and 50% in the patients aged >70 years. With increasing age, the diagnostic yield of EPS also increased. No significant differences of complication rate were found among the different age groups. It was concluded that EPS havd high diagnostic value in the patients with unexplained syncope. Its complications are few and mild. EPS may be recommended in elderly patients with unexplained syncope.

In order to investigate the changes of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in residual hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE), the expression levels of VEGF and bFGF expression in specimens surgically removed from 48 HCC patients were detected by immunohistochemical methods, and staining intensity of VEGF and bFGF was assessed by a computer-assisted image-analyzer. Among the 48 patients, 25 underwent partial hepatectomy alone (single operating group), and 23 were subjected to second stage surgical resection after TACE (TACE group). The results showed that the average absorbance value (A) of VEGF was higher in TACE group than that in single operating group (0.152±0.021 vs 0.131±0.012,P<0.01). The Average A of bFGF in TACE group was 0.127±0.023, higher than in single operating group (0.111±0.016,P<0.05). These results suggested that TACE of HCC can up-regulate the expression of VEGF and bFGF in HCC tissues possibly due to anoxia and ischemia.

The effects of estradiol and tamoxifen on the proliferation of estrogen receptor positive cells and the relationship between the tamoxifen tolerance and cell origin were investigated. The tissues of human endometrium and breast cancer were randomly selected following dissection for primary cell culture. After the breast cancer cells and endometrial cells were treated with 1×10⁻⁸ mol/L estradiol and/or 1×10⁻⁶ tamoxifen,³H-labelled thymine nucleotide was used to trace the kinetics of cell proliferation. There was no significant difference in the inhibition on the human endometrial cells between tamoxifen-treated group (6.3%) and control group (6.4%), but tamoxifen could significantly inhibit the proliferation of the human breast cancer cells (45.84%) as compared with control group (52.72%). Moreover, tamoxifen could significantly stimulate the proliferation of tamoxifen resistant breast cancer cells (9.64%) as compared with control group (6.32%). Estradiol could significantly stimulate the proliferation of all the three kinds of cells as compared with control group. The combined use of estradiol and tamoxifen could inhibit the proliferation of the endometrial cells and breast cancer cells as compared with estradiol used alone, but on the tamoxifen resistant breast cancer cells, they could more significantly stimulate the proliferation than E2. It was concluded that E2 could stimulate the proliferation of these three kinds of cells. However, the inhibitive effects of tamoxifen on the proliferation of these cells were dependent on the estradiol.

To investigate the effects of estrogen (E₂) on telomerase activity and its mechanism in human breast cancer cells, estrogen receptor positive MCF-7 cells were treated with different concentrations of E₂. Telomerase activity was measured by using TRAP-ELISA method, the cell cycle phases analyzed by using flow cytometry, and the expression of Cyclin D₁ detected by using immunohistochemistry method. The results showed that telomerase activity levels were increased in MCF-7 cells treated with 10⁻⁸ mol/L E₂ during the observed period (P<0.05), and E₂ increased telomerase activity levels in a dose-dependent manner (10⁻¹⁰-10⁻⁸ mol/L); Simultaneously, the cell cycle phases of MCF-7 cells treated with 10⁻⁸ mol/L E₂ were changed significantly: G₀/G₁ phase decreased from 60.52% to 50.93%, S phase increased from 29.03% to 30.83%; However, the expression of Cyclin D₁ was decreased. It was concluded that estrogen can upregulate telomerase activity of MCF-7 cells, and the effect can be blocked by antiestrogen tamoxifen. Its mechanism may be closely associated with modulation of cell cycle phases.

To study the expression of mTERT gene in the testis of SD rats and its significance, in situ hybridization (ISH) techniques were used to detect the expression of telomerase gene mTERT mRNA in the testis of SD rats. The expression of mTERT was detectable in different-age male SD rats testis. There was a positive correlation between the expression of mTERT and the location of germ cells (spermatogonia, spermatocyte, spermatid). In Sertoli cells, leydig cell and spermatozoa, telomerase mTERT was not detected. Type A spermatogonia expressed the highest level of telomerase mTERT mRNA. Our results suggest that the expression of mTERT gene in the testis of SD rats is of lifetime and coincide with the telomerase activity.

To study the expression of beta-human chorionic gonadotropin (βhCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of βhCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for βhCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of βhCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. β7 was most frequently transcribed subtype gene independent on the histology. These findings suggested βhCG gene transcription is not only involved in RCC but also in benign renal diseases.

In order to study the pathogenesis, clinical and pathological characteristics of proliferative lesions of the bladder, 50 cases of proliferative lesions of the bladder from 150 patients with complaints of frequency, urgency, hematuria and dysuria were subjected to cystoscopic biopsy of the suspicious foci in the bladder. In combination with the symptoms, urine routine and urodynamics, the relationship of proliferative lesions of the bladder to the inflammation and obstruction of the lower urinary tract was analyzed. Of the 50 cases of proliferative bladder lesions, 44 cases (88%) had lower urinary tract infection and 29 (58%) lower urinary tract obstruction. The patients with lower urinary tract obstruction were all complicated with infection. Three cases were associated with transitional cell carcinoma. Malignant cells were detected in 1 case by urinary cytologic examination. Proliferative lesions of the bladder, especially those without other obvious mucosa changes under cystoscopy, are common histological variants of urothelium in the patients with chronic inflammation and obstruction of the lower urinary tract. Chronic inflammation and obstruction of the lower urinary tract might be the causes for proliferative lesions of the bladder. It is suggested that different treatments should be applied according to the scope and histological type of the proliferative lesions.

To explore a new method for the therapy of the avascular necrosis of the femoral head, the recombinant plasmid pCD-hVEGF₁₆₅ was mixed with collagen and was implanted in the necrotic femoral head. The expression of vascular endothelial growth factor (VEGF) was detected by RNA dot hybridization and immunohistochemical method. The repair of the femoral head was observed by histological method. The results showed that the expression of VEGF was detectable in the femoral head treated with VEGF gene. Angiogenesis in these femoral heads was more abundant than the control. Bone repairing was augmented in the femoral head treated with VEGF gene. The results suggest that angiogenesis in bone tissue could be augmented by gene transfection of VEGF and bone repairing would be accelerated accordingly.

The feasibility of anterior lumbar intervertebral fusion with artificial bone in place of autogenous bone was investigated. Porous hydroxyapatite (HA)/ZrO₂ ceramics loading bone morphogenetic protein (BMP) were implanted after removal of lumbar vertebral disc in rabbits. The adjacent intervertebral discs were also removed by the same way and autogenous illic bone was implanted. SEM observation and biomechanical test were carried out. Compound bone had a bit lower osteoinductive activity than autogenous bone by SEM (Osteoindutive activity of artificial bone in 12 weeks was the same as that of autogenous bone in 9 weeks). Biomechanical test revealed that compound bone had lower anti-pull strength than autogeneous bone (P<0.001), but there was no significant difference in anti-pull strength between compound bone at 12th week and autogenous bone at 9th week (P>0.05). It was concluded that compound bone could, be applied for anterior spinal fusion, especially for those patients who can’t use autogenous bone.

The distribution and function of transforming growth factor-beta (TGF-β) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated. Twenty-four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8.0 mm in diameter through the whole bone. On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group. At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out. Immunohistochemical staining was performed for TGF-β assay by avidin-biotin complex method (SABC). Simultaneously, TGF-β was quantitatively analyzed by HPIAS-1000 imaging analysis system. The immunohistochemical staining for TGF-β revealed that osteoblasts and immature osteocytes highly expressed TGF-β. Diffused TGF-β positive staining particles appeared in the mesenchymal and fibrous-tissue. There was no significant difference in the TGF-β positive staining between two groups in the medial region to original osseous beds at different time points (P>0.05). However, in distal original osseous bed of the defected region, the positive expression of TGF-β in the Co/N-TCP group was significantly stronger than in the control group (P<0.05 or 0.01). The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-β to aggregate and precipitate the healing of bone defect.

The relationship between the hypertension and the aging process of hearing organ was investigated. Twenty Wistar 3-month old rats and 20 Wistar 12-month old rats, 20 spontaneously hypertensive rat stroke-prone (SHRSP) 3-month old rats and 20 SHRSP 12-month old rats free of middle ear infections as observed under otomicroscopy, with normal tympanic membrane and auricle reflex, were selected to be divided into two experimental groups and two control groups respectively. The tail artery blood pressure was measured non-invasively. The threshold of auditory brain-stem response (ABR) was measured by Spirit^™ evoked potential meter. The LDH and ChE staining in the inner ear was performed and the optical density was analyzed by the HPIAS analysis system. The results showed that there was no difference in the ABR thresholds, the activities of LDH and ChE between Wistar 3-month old group and SHRSP 3-month old group (P>0.05). The mean value of ABR threshold and the activities of LDH and ChE in the Wistar 12-month old group at relevant sections were significantly greater than those in the two 3-month old groups (P<0.05), whereas the mean value of ABR threshold and the activities of LDH and ChE in the SHRSP 12-month old group at relevant sections were significantly higher than those in the 3-month old control group (P<0.01). It was concluded that presbycusis existed in the Wistar 12-month old group rats. The glycogenosis and the abnormal secretion of neural transmitter were discerned after hypertension. All the above factors may worsen the aging of the hearing system.

The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49±15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89±10.13 IOD,P<0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR.

The role of catecholamine (CA) in the pathogenesis and development of macular edema of central serous chorioretinopathy (CSC) was studied, and its relations with visual acuity were investigated. Plasma concentrations of epinephrine (E) and norepinephrine (NE) were determined in 30 consecutive eyes with CSC. Central macular thickness analysis was done by RTA and all the data were compared with normal eyes and analyzed with SAS software package. Plasma concentrations of E and NE were increased to (569±123) ng/L and (721±104) ng/L respectively in active CSC patients, significantly higher than those in normal subjects (P<0.01), and decreased to normal in convalescent stage. RTA analysis revealed that the retinal thickness of CSC patients was increased at active and recovery stage as compared with normal subjects; and the plasma concentration of E was significantly correlated with central macular thickness (t=2.173,P<0.05). Also, central macular thickness measured by RTA was significantly correlated with the visual acuity (r=−0.8046,P<0.001) in CSC eyes. RTA analysis might be useful to quantitatively detect and evaluate prognosis in CSC patients. The plasma concentration of E, which was highly correlated with macular edema, might play an important role in the early damage and the pathogenesis of CSC.

To study the mechanism of Condyloma acuminatum (CA) recurrence, and the association of CA recurrence with the ability of the host derived lymphoblastoid cell lines (LCL) stimulated by LPS to produce tumor necrosis factor (TNF), EBV-transformed B LCL were used as TNF producing cells. The ability of LCL stimulated by LPS to produce TNF was measured by bioassay. The results showed that the LCL from CA patients (including recurrent and non-recurrent CA patients) produced similar level of TNF stimulated by LPS to that of normal controls (29.54%±11.28% vs 34.31%±11.46%,P=0.1498). The LCL of CA recurrent patients produced significantly lower amount of TNF than that of non-recurrent CA patients (23.72%±7.41% vs 37.33% ±11.10%,P=0.0032). Compared with the normal controls, CA recurrent patients showed a decreased ability to produce TNF (23.72%±7.41 vs 34.31%±11.46,P=0.0054), whereas CA non recurrent patients had the similar ability to the controls (37.33%±11.10 vs 34.31%±11.46,P=0.4914). It was concluded that the onset of CA was not relevant to the individual’s ability to produce TNF. But the recurrence of CA was associated with the ability to produce TNF. It was also indicated that the TNF involved cellular immunity might play an important role in the clearance of the residual HPV by the host after treatment.

To assess the potential therapeutic effect of propofol in the treatment of endotoxemia, 76 rats were randomly assigned to 5 groups: control group (A), endotoxemic group (B), pre-treatment group (C), simultaneous treatment group (D) and post-treatment group (E). Five h after endotoxin injection, PO₂, pH, MAP, plasma concentrations of Nitrite/nitrate (NO₂⁻/NO₃⁻) and mortality rates were assessed in each group. After the rats were sacrificed, lung tissue was sampled to measure myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-α contents. It was found that endotoxin injection produced progressive hypotension, metabolic acidosis, and a large increase in the plasma NO₂⁻/NO₃⁻ concentrations and increased mortality rates in 5h. Endotoxin injection significantly increased MPO activity and TNF-α contents in lung tissue (P<0.01 orP<0.05). These changes response to endotoxin were significantly attenuated in the groups B, C and D. But these beneficial effects were blunted in the group E. The results suggest that propofol administration may offer advantages in endotoxemia.

A series of imaging features of extranodal, multi-systemic involvements in Non-Hodgkin’s lymphoma (NHL) were investigated. The clinical data and imaging findings of 16 patients with pathologically proved NHL were retrospectively analyzed. The related literatures were reviewed. Of the 16 cases of NHL, skeletal involvement was found in 4, nasal cavity and nasal sinuses were involved in 4, too. Lesion in the thorax was seen in 3 patients, hepatic involvement occurred in one case, cerebral ventricle was affected in 3 cases, mesentery was involved in one case. Even though extranodal involvement of NHL exhibited extremely variable patterns, there were some relatively typical imaging findings. Emphasized in this report were the relatively specific imaging manifestations of different systems, which may mimic infectious or other neoplasms of different sites. The importance of imaging studies lies in the availability for diagnosis, staging and follow-up of NHL. Combined with the clinical and other related information, the diagnostic accuracy can be further improved, thus, providing reliable evidence in guiding clinical management.