Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of humanMycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from humanM. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP HSP65. Results of restriction endonuclease analysis. PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69% in total bacterial protein and 74.09% in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody againstM. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.

To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal adenocarcinoma (CA), immunohistochemical SP method was used to detect expressions of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 in paraffinembedded tissues of 18 cases of MEM, 20 OA and 20 CA. The results showed that there was a significant difference in the expressions of E-cadherin, CA19-9 and MOC-31 between MEM and OA group (P<0.05). Similarly, the difference in the expression of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 between MEM and CA groups is significant (P<0.05). These results indicate that HBME-1 could be used as a positive marker in distinguishing MEM from CA. E-cadherin, CA19-9 and MOC-31 are considered to be useful negative markers in diagnostic distinction between MEM and metastatic adenocarcinomas, including OA and CA. CK7 is the best positive marker in distinguishing MEM from CA, but this marker appears to be valueless in discriminating MEM from OA.

This study pharmacokinetically examined the lovastatin sustained-release tablet and sustained-release capsule in Beagle dogs. An reversed-phase HPLC method was established for the determination of lovastatin in Beagle dog plasma. Pharmacokinetic findings were compared among three preparation (lovastatin sustained-release tablet, T_p; sustained-release capsule, T_j and conventional capsule). Our results showed that the pharmacokinetic parameters in 6 dogs after single-dose oral administration of three perparations were calculated. T_max, C_max and MRT revealed significant difference (P<0.05). Relative bioavailability was 111.5±16.9% (T_p) and 110.4%±9.6% (T₁). The pharmacokinetic parameters in the 6 dogs after multiple-dose oral administration of three perparations, T_max, C_max MRT and DF had significant difference (P<0.05); C_av, C_min and AUC_0–24h displayed no significant difference (P>0.05). It is concluded that the lovastatin sustained-release tablet and sustained-release capsule are able to maintain a sustained-release for 24 h.

To study the effects of cyclooxygenase 2 selective inhibitor Nimesulide (NIM) combined with Cisplatin (DDP) on human lung cancer and the possible mechanisms, the proliferation and appoptosis of human lung cancer cell line A549 were evaluated by MTT reduction assay and flow cytometry respectively. The inhibitory effect on neoplasiain vivo was tested on nude mice subcutaneously implanted tumor. Our results showed that NIM and DDP could inhibit A549 cell proliferation in a concentration-dependent pattern; this action was enhanced when NIM (25 μmol/L) was given in combination with DDP and they worked in a synergistic or additive pattern as DDP concentration ≥1 μg/ml. NIM and DDP could induce A549 cells apoptosis and the action was augmented when used in combination (P<0.01). NIM and DDP could inhibit the growth of subcutaneously implanted tumors on nude mice (P<0.05,P<0.01) and the inhibitory rate of NIM combined with DDP was significantly higher than that of NIM or DDP group (P<0.01,P<0.01). It is concluded that combined use of NIM and DDP has significant synergistic antitumor effects on lung cancer cell line A549 and in animalsin vivo. The synergy may be achieved by growth inhibition and apoptosis induction.

To investigate the expression of hypoxia inducible factor 1-alpha (HIF-1α) and its correlation with P53 and vascular endothelial growth factor (VEGF), immunohistochemical technique was employed to detect the protein expressions of HIF-1α, P53 and VEGF in specimens from 57 patients with lung cancer. The results indicated that the total positive proportion of HIF-1α expresion was 63% and the HIF-1α expression was more frequent in bronchiole-alveolar carcinoma (86%) than in other lung cancer. There was a strong association of HIF-1α with VEGF and P53 protein expressions. It is concluded that HIF-1α overexpression is a common event in lung cancer, which may be related to the up-regulation of the angiogenic factor VEGF and oncogene mutant P53 protein.

To elucidate the mechanism of arrhythmia in healed myocardial infarction (HMI), the changes of action potential duration (APD), transient outward potassium current (1_ω), delayed rectifier potassium current (I_k) and inward rectifier potassium current (I_ki) of left ventricular myocytes in non-infarcted zone of HMI were investigated. Rabbits were randomly assigned into two groups: HMI group, in which animals were subjected to thoracotomy and ligation of the circumflex coronary and sham-operated group, in which rabbits underwent thoracotomy but no conorary ligation. 3 months after the operation, the whole myocyte patch clamp technique was used to record APD, I_to I_k, and I_ki of ventricular myocytes in non-infarcted zone. Our results showed that the membrane capacitance was larger in HMI group than in sham-operated group. Action potential duration was significantly lengthened in HMI group and early afterdepolarization (EAD) appeared in HMI group. The densities of I_to, K_{k, tail}, and K_ki were reduced significantly in HMI group, from 6. 72±0.42 pA/pF, 1.54±0.13 pA/pF and 25.6±2.6 pA/pF in sham-operated group to 4.03±0.33 pA/pF, 1.14±0.11 pA/pF and 17.6±2.3 pA/pF, respectively. It is concluded that the reduced densities of I_to, I_k.tail and I_ki in ventricular myocytes of non-infarcted zone in HMI were responsible for the prolongation of APD and the presentation of EAD which played important roles in the development of malignant arrhythmia in HMI.

This study evaluated the effects of adenovirus vector mediated human vascular endothelial growth factor-165 (hVEGF₁₆₅) gene on prevention of restenosis after angioplasty. Rabbit models of bilateral carotid artery injury were established by balloon denudation. The recombinant adenoviruses containing hVEGF₁₆₅ cDNA was directly injected into left side of the injured carotid arteries. On day 3 and week 3 after transfection the expression of VEGF was observed by RT-PCR and immunohistochemistry. The thrombokinesis, reendothelialization (rET) and intimal hyperplasia in carotid arteries were evaluated by computerized image analysis system 3 weeks after gene transfer. The changes in the VEGF gene-treated side were compared with the control side. Our results showed that 3 days and 3 weeks after hVEGF₁₆₅ gene transfer the VEGF mRNA and antigen expression were detectedin vivo. 3 weeks after the transfer, the carotid artery rET was markedly better in the VEGF gene-treated group compared with the control. The thrombokinesis, intima area/media area (I/M), maximal intimal and medial thicknesses (IT_max and MT_max) demonstrated a statistically significant decrease in arteries treated with VEGF gene as compared with the control group. It is concluded that VEGF gene transfer could be achieved by intra-arterial injection of recombinant adenoviruses. It might accelerate the restoration of endothelial integrity, inhibit thrombokinesis and attenuate intimal hyperplasia in the injured arteries after VEGF gene transfer. This procedure could be useful in preventing restenosis after angioplasty.

To investigate the mechanism and the suppressive effect of human cytomegalovirus (HC-MV) on colony forming unit-megakaryocyte (CFU-MK), semi-solid culture system was used to observe the effect of HCMV AD169 strain on CFU-MK's growth of 18 cord blood samples. HCMV DNA and immediate early (IE) protein mRNA in CFU-MK was detected by PCR and reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that HCMV AD169 significantly suppressed the formation of CFU-MKin vitro. Compared with the mock group, the CFU-MK colonies decreased by 21.6%, 33.8% and 46.3%, respectively, in all the 3 infected groups (P<0.05), suggesting the suppression and the titer of the virus was dose-dependent. Both HCMV DNA and the expression of HCMV IE protein mRNA were positively detected in the colony cells of viral infected group,. It is concluded that HCMV AD169 strain could inhibit the differentiation and proliferation of CFU-MK by directly infecting their progenitors. There was early transcription of HCMV IE protein in CFU-MK infected by virus.

To investigate the effect of atorvastatin on lipid metabolism in type 2 elder diabetes patients with hyperlipidemia, 26 patients with type 2 elder diabetes complicated with hyperlipidemia were treated with atorvastatin (10 mg/d) for 8 weeks. The serum triglyceride (TG), high density protein cholesterol (HDL-C) and low density protein cholesterol (LDL-C) were measured before and after the treatment. Meanwhile, the non-denaturing polyacrylamide gradient gel electrophoresis was used for detection of small-sized LDL (SLDL). Our results showed that TG dropped from 4.88±0.72 mmol/L to 2.65±0.32 mmol/L; HDL-C was increased from 0.85±0.31 mmol/L to 1.28±0.29 mmol/L; LDL-C was declined from 3.71±2.98 mmol/L to 2.10±1.22 mmol/L, sLDL-A was increased from (42.49±8.1)% to (53.27±7.5)%; LDL-B was decreased from (57.91±8.1)% to (46.73±7.5%) (P<0.05). The level of blood glucose was not changed at the end of 8th week. It is concluded that atorvastatin has satisfactory lipid-regulating effects on type 2 elder diabetes patients with hyperlipidemia.

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1 genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTCin vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10⁻³ mol/L, 10⁻⁴ mol/L carbachol) could induced a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10⁻³ mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10⁻³ mol/L carbachol)in vitro for 30 min. Either M3 cholinergic receptor antagonist (10⁻⁵) mol/L atropine) or NF-κB inhibitor 10⁻² mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1 genes in pancreatic acinar cellsin vitro through the activation of NF-κB.

To investigate TLR2 (Toll-like receptor 2) mRNA expression in ischemic hepatic lobes under the condition of partial hepatic ischemia/reperfusion injury in BALB/c mice and its relationship with liver function impairment. A partial ischemia/reperfusion injury model was established. The portal vein and hepatic artery supply to the median and left lobes of the liver were obstructed by an atraumatic artery micro-clip, with the obstruction lasting for about 60 min. Then reperfusion was fulfilled by removal of the clip. The liver samples were collected at the 4th h after the restoration of blood inflow. Total RNA was extracted from the liver samples and analyzed quantitatively by method of real-time PCR. At the same time, portal vein serum and plasma were taken respectively for further detection of the level of endotoxin, tumor necrosis factor alpha (TNF-alpha) and plasmic alanine aminotransferase (pALT). The results indicated that TLR2 mRNA in ischemic lobe was up-regulated markedly in mice partial liver ischemia/reperfusion injury model compared to that in sham operation group (ΔCt: 1.05±1.02 vs 5.08±1.36,P<0.001). The level of portal vein pALT and TNF-alpha increased significantly (112.32±17.56 pg/ml vs 6.07±5.33 pg/ml,P<0.01; 890±127 μ/L vs 30±5 μ/L,P<0.001). However, the level of portal vein endotoxin remained below the normal line, suggesting a state of non-endotoxemia. TLR2 mRNA expression in ischemic lobe, as well as portal vein pALT and TNF-alpha, was up-regulated in the model of mice partial ischemia/reperfusion injury, suggesting the involvement of TLR2 in ischemia/reperfusion pathological process.

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made.In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3–7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM).In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culturein vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. Thein vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.

To investigate the expression of Bcl-2 and Bcl-xl gene in sensitive (A2780) and drug-resistance (AD6) human ovarian cancer cell lines and explore the molecular mechanism of multidrug resistance. A2780 and AD6 were detected by using DNA gel electrophoresis, flow cytometry and RT-PCR. Our results showed that (1) “DNA ladder” was observed in A2780 and AD6 after cisplatin treatment; (2) after 3.0, 6.0, 9.9 μg/ml of cisplatin treatment, a significant difference was noted in the rate of apoptosis between in A2780 and AD6 (P<0.05); (3) Bcl-2 and Bcl-xl genes were overexpressed in AD6. After cisplatin treatment, the expression of Bcl-2 and Bcl-xl genes was down-regulated in A2780 and AD6. It is concluded that cisplatin could induce the apoptosis of ovarian cancer cells, and the over-expression of Bcl-2 and Bcl-xl genes may contribute to apoptotic inhibition and the development of multidrug-resistance of human ovarian cancer.

To compare the diagnostic value of soluble intercellular adhesion molecule 1 (sICAM-1) with that of c-reactive protein (CRP) for detecting chorioamnionitis (CAM) in serum of women with premature rupture of membranes (PROM), 55 pregnant women with PROM, including 18 pregnant women with preterm premature rupture of membranes (PPROM) and 20 normal pregnant women at term (TPROM) were studied. Maternal serum were measured by Sandwish enzyme-linked immunoabsorbent assay (ELISA) for sICAM. CAM was histologically confirmed after delivery. The results revealed that (1) maternal serum levels of sICAM-1 and CRP were significantly higher in women with PROM than those without it: (2) maternal serum levels of sICAM-1 and CRP were significantly higher in women with CAM than those without it: (3) serum levels of sICAM-1 in PPROM women were similar to those in TPROM women, whereas serum levels of CRP in PPROM women were significantly higher than those in TPROM women; (4) the sensitivity, specificity, positive predictive value, negative predictive value, Kappa index and area under receiver operating characteristic (ROC) curve of maternal serum sICAM-1 (cutoff 104.7 ng/ml) and CRP (cutoff 1.03 mg/dl) for diagnosing CAM were 100%, 91.2%, 87.5%, 100%, 0.20, 0.995 and 81.0%, 73.5%, 65.4%, 86.2%, 0.13, 0.811, respectively; (5) among the mild histological CAM group, severe histological CAM group and clinical CAM group, the difference in maternal serum levels of sICAM-1 were significantly (P<0.001), with the order of concentration from high level to low level corresponding to the severity of CAM. It is concluded that maternal serum level of ICAM-1 is superior to that of CRP as biomarker for diagnosing intraamniotic infection in pregnant women with PROM.

This study investigated the relationship between angiotensin-converting enzyme (ACE) gene insertion/deletion polymorphism and the occurrence, severity, prognosis of HSPN. The polymorphism of ACE gene in 103 HSPN cases and 100 healthy children was studied by using the polymerase chain reactions (PCR). Its relation to the clinical manifestation, pathological classification and prognosis of HSPN was analyzed accordingly. The results showed that: (1) there was a significantly higher frequency for DD genotype in HSPN children (P<0.01); (2) DD genotype was more frequently seen in HSPN children with gross hematuria and massive proteinuria (P<0.05), while DI genotype was more common in HSPN children group with renal insufficiency (P<0.05); (3) although mesangial proliferative lesion was most frequently observed in 21 biopsied HSPN children, and DD genotype frequency was still higher in children with severe pathology (Class III IV); (4) II genotype was significantly frequent in HSPN children with complete remission in the follow-up of 32 HSPN children. It was concluded that the deletion allele of ACE gene might play a role, at least to some extent, in the occurrence, deterioration and progression in juvenile HSPN.

To explore the relationship between Insulin-like growth factor (IGF)-I, −II and lung development in neonatal rats. 80 timed pregnant Sprague-Dawley (SD) rats were randomly divided into 4 groups (n=20): group A (Control group), group B (Dexamethasone (DEX) 1 group), group C (DEX 2 group), group D (retinoic acid (RA) group). 20 pregnant rats in group A, B and D were injected subcutaneously or intraperitoneally with vehicle (NS), DEX, or RA respectively during gestational day 16 to 18. All newborn rats in group C were subcutaneously injected with DEX at day 1 to 3 after birth. The lung tissue was obtained at the following times: fetuses at gestational ages of 18, 20 and 21 days, and 1, 3, 5, 7, 10, 14 and 21 days after birth. Lung tissues were used for histopathological study, the polypeptides analysis of IGF-I, −II (immunohistochemistry and Western blot) and mRNA analysis (RT-PCR). The results showed that the strongest expression of IGF-I in group A and D occurred at ages of 5–7 days (alveolar stage). The stronger their expressions, the better the alveolar develop. The peak stage of expression in group B occurred earlier, on the day 3 after birth. Compared with group A, the expression of IGF-I during gestation age of 18 days to age of 3 days in group B were significantly higher (P<0.01), but significantly lower at other time points (P<0.01). The expression of IGF-I was lower in group C all the time and always higher in group D than those in group A (P<0.01). The peak expression of IGF-II took place at the gestation age of 18 days, then gradually dropped to trace. During 18 days of gestation to age of 3 days, the expression IGF-II in group B was significantly higher than that in group A (P<0.01). No difference was found among all other groups. The change in the expression of IGF-I, −II mRNA in all 4 groups was similar to that of their polypeptides. The results suggested that there is a close linking between IGF-I, −II and lung development in newborns. The IGF-II works at early stage and the that of IGF-I works at the stage of new septa formation and alveoli maturation. The stronger their expressions, the more mature the lung development.

To evaluate the implication of methymalonic acid (MMA) in the early diagnosis of neural tube defects (NTD), a quantitative assay for MMA was established by using gas chromatographymass spectrometry with stable isotope of MMA as an internal standard. Amniotic fluid and maternal urine MMA concentration, maternal serum folate, red blood cell folate and vitamin B₁₂ levels were measured in the middle term of NTD-affected and normal pregnancies. Amniotic fluid and maternal urine MMA concentrations in the middle term of NTD affected pregnancies (1.4±0.9 μmol/L, and 22.1±12.6 nmol/μmol creatinine) were significantly higher than that of normal pregnancies (1.0±0.4μmol/L, and 2.5±1.1 nmol/μmol creatinine). There was no significant differences between normal and NTD pregnancies for serum folate, red blood cell folate and vitamin B₁₂ levels. The results suggested that MMAs in amniotic fluid and maternal urine are sensitive markers for early diagnosis of NTD. Vitamin B₁₂ is an active cofactor involved in the remethylation of homocycteine and its deficiency is an independent risk factor for NTD. MMA is a specific and sensitive marker for intracellular vitamin B₁₂ deficiency. This study suggests that it is necessary to monitor the vitamin B₁₂ deficiency and advocates vitamin B₁₂ supplementation with folate prevention program.

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA. The increased ratio of Bcl-2 and Bax mRNA may contribute to the anti-apoptic effect of nimodipine. The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new strategy to manage neuronal damage.

To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs)in vitro, and to evaluate the inductive role of hemin on his pathway, HTMCs of the third to fourth generation were culturedin vitro. Reverse transcipase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.

To explore the biological effects of light on trabecular cells, cultured bovine trabecular cells were exposed to visible light of different wavelength with different energy. Cellular morphology, structure, proliferation, and phagocytosis were observed. The cells showed no remarkable changes when the energy was low. When the exposure energy reached 1.12 mW/cm², the cytoplasm showed a rough appearance, and cell proliferation and phagocytosis decreased. This photoxicity was strong with white light (compound chromatic light), moderate with violet light or yellow light, and mild with red light.

An experimental model of rhegmatogenous retinal detachment (RRD) in rabbits was established to simulate the pathophysiologic condition of human RRD, 24 rabbits were randomly divided into 3 groups and underwent vitrectomy with a vitrector and/or retinotomy with a Charles flute needle, with 12 in group I (vitrectomy and retinotomy), 7 in group II (retinotomy) and 5 in group III (vitrectomy). All animals underwent follow-up examinations with direct and indirect ophthalmoscopy and fundus photography 12 h and day 1, 3, 5, 7, 10, 14, 21, and 28 after the procedure(s). Retinal changes were recorded. As a result, 10 RRDs were successfully established in group I. Direct and indirect ophthalmoscopy and fundus photography demonstrated typical features of RRD. No RRD developed in group II and III. It was concluded that the experimental rhegmatogenous retinal detachment produced in a rabbit model after vitrectomy with retinotomy in this study was a convenient and reliable one. This RRD model mimicked the typical pathophysiological changes in humans.

To assess the left ventricular regional relaxation abnormalities in patients with hypertrophic cardiomyopathy (HCM) by quantitative tissue velocity imaging (QTVI), Doppler echocardiography and QTVI were performed in HCM (n=10) and healthy subjects (n=11) at apical longaxis, two-chamber and four-chamber views. Regional early diastolic velocity (rVe) and regional atrial contraction (rVa) were measured at each segment of ventricular middle, basal and annular levels. Mean rVe and mean rVa at three levels as well as mean rVe/rVa ratio were calculated. Our results showed that transmitral inflow peak velocities during early diastole (E) and atrial contraction (A) were also measured and E/A ratio was calculated. The rVe of all left ventricular segments in HCM were lower than those in healthy subjects (P<0.05), but compared with healthy subjects majority of rVa in HCM were not different except inferior wall and anterior wall. E between HCM and healthy subjects was different (P=0.036), while mean rVe between them was significantly different (P<0.0001). Mean rVa and mean rVe/rVa of three levels were lower in HCM than in healthy subjects (P=0.05), but there were no differences in A and E/A between them (P=0.22,P=0.101). Left ventricular regional myocardial relaxation is reduced in HCM. Transmitral inflow E and A are influenced by preload, relaxation of myocardium and atrial contraction, etc., while rVe and rVa reflect myocardial relaxation function independently. QTVI is more sensitive and more accurate than conventional Doppler imaging for characterizingregional diastolic properties in HCM.

To examine the role of nitric oxide in the β-adrenergic vasodilation of epicardial coronary arteries in dogs, 12 dogs were instrumented for measurement of left anterior descending coronary artery diameter by transthoracic echocardiography before and after dobutamine (5 μg/kg/min IV) with and without intracoronary infusion of N^G-monomethyl-L-arginine (L-NMMA) (1 mg/kg). In all 12 dogs, the diameter of left anterior descending coronary artery increased significantly from 2. 35±0.25 mm to 2.59±0.24 mm (P<0.001) after dobutamine administration. In 6 of the 12 dogs, the percent change in left anterior descending coronary artery diameter induced by dobutamine decreased significantly from 12.5%±8.6% to −1.5%±5.4% (P<0.05) after the administration of intracoronary L-NMMA (1 mg/kg for 5 min) to block nitric oxide synthesis from L-arginine. The study demonstrated that nitric oxide formation contributes to the β-adrenergic dilatory response of epicardial coronary arteries to dobutamine in dogs.

To investigate the value of echocardiography in the diagnosis of total anomalous pulmonary venous connection (TAPVC), 16 patients in our hospital were diagnosed to have TAPVC by echocardiography from year 1994 to 2001. In 11 cases the results of echocardiography were compared to those of surgery. Each patient was examined by using a combination of precordial, suprasternal and subcostal windows to visualize all the pulmonary veins and their drainage sites, common pulmonary venous trunk, and other associated abnormalities. Of the 16 cases, the drainage sites were as follow: supracardiac in 10, via vertical vein in 9, directly to superior vena cava in 1; cardiac in 5, via coronary sinus in 2, directly to right atrium in 3. Diagnoses were correctly made in all the 11 cases as confirmed by surgery. Echocardiography can also assess pulmonary arterial pressure and detect other associated abnormalities. It is concluded that echocardiography is the preferred examination method in the diagnosis of TAPVC before surgery. With careful examination using multiple windows and sections, TAPVC can be accurately diagnosed by echocardiography.

The aim of this study was to determine the role of neutrophil collagenase in the pathogenesis of acute lung injury induced by endotoxin. 28 Sprague-Dawley were randomized into control group and LPS-enduced groups. Samples of left lung were obtained in 2 h (group L₁), 6 h (group L₂), 12 h (group L₃) after intravenous LPS. Immunohistochemsitry was employed for detection of expression of neutrophil collagenase. Pathological scores, lung wet/dry weight ratio and the number of neutrophils were measured. The results showed that the concentration of neutrophil collagenase in LPS-enduced groups (group L₁, L₂, L₃) were significantly higher than that of control group (P<0.01). Pathological scores, lung wet/dry weight ratio and the number of neutrophils in LPS-enduced groups (group L₁, L₂, L₃) were also significantly higher than that of control group (P<0.01). Moreover, among group L₁, L₂ and L₃, there were significant correlations in concentration of neutrophil collagenase and pathological scores, lung wet/dry weight ratio, the number of neutrophils (P<0.05). The present study showed that neutrophil collagenase play an important role in the pathogenesis and progress of endotoxic acute lung injury.

To evaluate the relationship between erythrocyte injury and intracellular calcium ion overload, and the protective effect of propofol on erythrocytes during cardiopulmonary bypass (CPB). 40 children with congenital heart diseases who underwent surgical repair under CPB were included. The patients were randomly divided into two groups: control group (group C) and propofol group (group P). Anesthesia was maintained in the patients with 6 mg/kg/h propofol in Group P, and those in the Group C inhaled 1%–2% isoflurane. The blood samples were taken before CPB, 30 min after CPB, at the end of CPB, and 2 h and 24 h after CPB to measure the content of erythrocyte intracellular calcium ion (E−Ca²⁺), Ca²⁺−Mg²⁺-ATPase and Na⁺−K⁺-ATPase activities, index filtration of erythrocytes (IF), mean corpuscular volume (MCV) and the concentration of plasma free hemoglobin (F−Hb). Results showed that in the control group, E−Ca²⁺, IF, MCV and F−Hb were gradually increased and Ca²⁺−Mg²⁺-ATPase and Na⁺−K⁺-ATPase activities were decreased. The increase of E−Ca²⁺ was linearly paralleled to IF, MCV and F−Hb. In propofol group, all the above-mentioned parameters were significantly improved (P<0.05). This study suggests that erythrocyte injury is related to elevation of intracellular calcium during CPB and propofol has a protective effect on erythrocyte injury.

The compounds from the root ofRhododendron molle G. Don were isolated, purified by various chromatographic techniques, and their structures were identified according to the physical and chemical features and spectral data. Three compounds were separated from the root ofRhododendron molle G. Don and identified as Rhodojaponin-III,taraxerol, β-sitosterol for the first time.