To find a technique of detecting and differentiating enteric adenoviruses (EAds) in clinical samples, a novel PCR approach was developed. EAds were able to be detected by use of a pair of subgroup F general primers (P1 and P2), and they were also be able to be differentiated from each other in the presence of another adenovirus type 40 (Ad40) specific primer (P3) in the same tube. Our results showed that there was one band for Ad41 and two bands for Ad40, respectively, on running-gel after PCR performance. PCR was performed on 40 specimens in parallel directly with dot-hybridization assay on the same diluted stool samples. 20 of 40 specimens were positive by hybridization (of them 12 were Ad41 and 8 were Ad40), whereas 26 were positive by PCR performance on the same samples with Ad41 18 and Ad40 8 positive as well. Our study indicated that this novel method could be used in clinical laboratory or in epidemic investigation for Eads

By using AdEasy system, which is based on the homologous recombination in bacteria, an EGFP labeled recombinant adenovirus vector containing hVEGF165 was constructed quickly and efficiently expressed in mouse haematopoietic cells, First, hVEGF165 coding sequence was subcloned into shuttle plasmid pAdTrack-CMV, then cotransformed with adenoviral backbone vector pAdEasy-1 into E. coli strain BJ5183. The recombinant adenoviral plasmid was transfected into HEK293 cells to assembly replication-defective adenovirus Ad-EGFP/hVEGF165. The expression of EGFP could be easily detected. The rate of EGFP positive mouse bone marrow mononuclear cells by flow cytometric analysis was 27.3% (MOI=100), and the expression of hVEGF165 protein in the supernatant was (1385±332) pg/10⁶ cells. These results suggest that the construction of adenovirus vector by homologous recombination in bacteria features high efficiency and simplicity. The prepared high titer Ad-EGFP/hVEGF165 can be used an efficient helpful vector to infect hematopoietic cells.

To explore the effects of ligustrazine on bone marrow heparan sulfates (HS) expression in bone marrow transplantation (BMT) mice, the syngeneic BMT mice were orally given 2 mg ligustrazine twice a day. On the 7th, 10th, 14th, 18th day after BMT, peripheral blood cells and bone marrow nuclear cells (BMNC) were counted, and the expression levels of HS in bone marrow and on the stromal cell surfaces were detected by immunohistochemistry and flow cytometry assay respectively. In ligustrazine-treated group, the white blood cells (WBC) and BMNC on the 7th, 10th, 14th, 18th day and platelets (PLT) on the 7th, 10th day were all significantly more than those in control group (P<0.05). The bone marrow HS expression levels in ligustrazine-treated group were higher than those in control group (P<0.05) on the 7th, 10th, 14th, 18th day. However, the HS expression levels on the stromal cell surfaces showed no significant difference between the two groups on the 18th day (P>0.05). It was concluded that ligustrazine could up-regulate HS expression in bone marrow, which might be one of the mechanisms contributing to ligustrazine promoting hematopoietic reconstitution after BMT.

The effects of heparin on the expression of transforming growth factor-β₁ (TGF-β₁) and two extracellular matrix components laminin (LN) and fibronectin (FN) in diabetic rat glomeruli were investigated. Twenty-six rats were randomly divided into control group (C,n=8), diabetic group (D,n=9), and diabetes+heparin group (DH,n=9). After 8-week therapy of heparin (200 U once daily by abdominal injection), TGF-β₁, LN and FN expression in glomeruli was detected by immunohistochemical method. The results showed that the expression levels of TGF-β₁, LN and FN were higher in group D than in group C. It was found that heparin could reduce 24-h urinary albumin excretion and inhibit overexpression of TGF-β₁, LN and FN in glomeruli of diabetic rats. It suggested that the inhibitory effect of heparin on diabetic glomerular sclerosis was at least partly related with the inhibition of TGF-β₁ expression.

To investigate the molecular mechanism of leptin regulating insulin secretion through determining the regulation of insulin secretion and the insulin receptor substrate (IRS)-2-associated phosphoinositide 3-kinase (PI3K) activity by leptin in the isolated rat pancreatic islets, pancreatic islets were isolated from male SD rats by the collagenase method. The purified islets were incubated with leptin 2 nmol/L for 1 h in the presence of 5.6 mmol/L or 11.1 mmol/L glucose. Insulin release was measured using radioimmunoassay. IRS-2-associated activity of PI3K was determined by immunoprecipitate assay and Western blot. The results showed that in the presence of 5.6 mmol/L glucose, leptin had no significant effect on both insulin secretion and IRS-2-associated PI3K activity, but in the presence of 11.1 mmol/L glucose, insulin release was significantly inhibited after the islets were exposed to leptin for 1 h (P<0.01). PI3K inhibitor wortmannin blocked the inhibitory regulation of leptin on insulin release (P<0.05). Western Blot assay revealed that 2 nmol/L leptin could significantly increase the IRS-2-associated activity of PI3K by 51.5% (P<0.05) in the presence of 11.1 mmol/L glucose. It was concluded that Leptin could significantly inhibit insulin secretion in the presence of 11.1 mmol/L glucose by stimulating IRS-2-associated activity of PI3K, which might be the molecular mechanism of leptin regulating insulin secretion.

The effects of intensive versus regular therapy on incidence and progress of microalbuminuria in type 2 diabetes were compared. During a follow-up of 3 years, 96 cases of diabetes mellitus were randomized to intensive and regular therapy groups. HbA_1c goal was same in the two groups, but the goal of blood pressure (Bp) and lipid was more strict in the intensive therapy group than in the regular therapy group. There was statistically significant difference in the incidence and progression of vascular complications between the two groups. Logistic stepwise-regression analysis (odds ration, OR) showed that there was significant difference in the progression of nephropathy (OR 0.24, 95% CI 0.12–0.76), retinopathy (OR 0.38, 95% CI 0.16–0.88), peripheral neuropathy (OR 0.42, 95% CI 0.22–0.86) and autonomic neuropathy (OR 0.29, 95% CI 0.12–0.86) between the two groups (P<0.01). It was concluded that intensive blood glucose controlling could retard diabetic vascular complications. Intensive therapy of multiple factors interventions (controlling Bp, regulating blood lipid, improving microcirculation) could decrease various risk factors for diabetic vascular complications.

To investigate the expression of NOS III mRNA and protein in cultured porcine cerebral arterial endothelial cells (CAEC) during hypoxia and reoxygenation and the effects of L-Tetrahydropalmatine (L-THP) on the gene expression of NOS III in CAEC during hypoxia and reoxygenation. The cultured CAEC were divided into 5 groups; control, hypoxia, hypoxia+reoxygenation, hypoxia +L-THP and reoxygenation +L-THP groups. NOS III mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemistry was used to detect the level of NOS III protein. The expression of NOS III mRNA and protein were increased when CAEC were exposed to hypoxia for 1 h, and significantly decreased during reoxygenation 2, 6 and 12 h after 1-h of hypoxia. L-THP from 10⁻⁸ mol/L to 10⁻³ mol/L could inhibit the up-regulation of NOS III gene expression during hypoxia and down-regulation of NOS III gene expression during reoxygenation.

To study the relationship between the disturbance of nitric oxide/endothelin-1 (NO/ET-1) and the hepatic ischemia/reperfusion (I/R) injury as well as the regulation of the NO/ET-1 system by the hepatic ischemic preconditioning (IPC), the changes of the NO/ET-1 system and their relationship with the hepatic I/R injury were compared between the I/R group and the IPC+I/R group in a rat hepatic I/R model. 2 h after reperfusion, the liver tissues were examined for expressed inducible nitric oxide synthase (iNOS) mRNA by RT-PCR. In the acute phase of hepatic reperfusion, the ratio of NO/ET-1 was reduced, which was due to the significant reduction of NO₂⁻/NO₃⁻ (the metabolic product of NO) and significant elevation of ET-1 in the blood plasma. The content of ALT, AST, LDH and TNF-α in blood plasma, and level of MDA in liver tissue were increased but ATP in liver tissue was reduced, and the hepatic damage was deteriorated. The protection of the hepatic IPC was associated with the elevated ratio of NO/ET-1 caused by the elevation of NO₂⁻/NO₃⁻, and reduction of ET-1 as well. No iNOS mRNA was detected in the liver tissues. It was concluded that hepatic I/R injury was related to the disturbance of NO/ET-1. The protection of the hepatic IPC in the acute phase might be mediated by its regulation of NO/ET-1 system. The cNOS rather than the iNOS generated the NO in this scenario.

The effect of thyrosine kinase, calmodulin and voltage-dependent Ca²⁺ channel on the proliferation of hepatoma cells induced by EGF was studied. Hepatoma cell line SMMC7721 was cultured in RPMI1640 serum-free medium. DNA synthesis rate of hepatoma cells was measured by H-TdR incorporation. 10⁻⁹ mol/L EGF could significantly stimulate the proliferation of hepatoma cells (P<0.05), and this effect might be significantly inhibited by tyrosine kinase inhibitor (P<0.001). Calmodulin inhibitor W-7 had no effect on the basic phase of cultured hepatoma cells (P>0.05), but it had very significantly inhibitory effect on the proliferation of hepatoma cells induced by EGF (P<0.001). Voltage-dependent Ca²⁺ channel inhibitor Varapamil had no inhibition on the proliferation of hepatoma cells induced by EGF (P>0.05). It had no effect on the basic phase of cultured hepatoma cells, (P>0.05). It is suggested that tyrosine kinase and Ca²⁺-calmodulin-dependent pathway may play a critical role on the proliferation of heptoma cells induced by EGF and voltage-dependent Ca²⁺ channel is independent of the effect of EGF.

To study the effect of paroxetine on depressive symptom accompanying somatic disease and the value of platelet 5-HT concentration in the diagnosis of depression, 30 patients with depressive symptom were treated with paroxetine. All patients were evaluated on Zung and HAMD scale and assayed of platelet 5-HT concentration before and after treatment. It was found that patients had a lower level of platelet 5-HT concentration than healthy people (P<0.01). After six weeks of treatment, depressive and somatic symptoms were both improved (P<0.01) and platelet 5-HT concentration was even lower (P>0.05). It was suggested that paroxetine was a good antidepressant and platelet 5-HT concentration was useful in the screening of depression.

The expression of an auto-antibody in patients with edematous acute pancreatitis and its possible clinical significance was investiaged. Eighteen cases of acute pancreatitis were chosen as experimental group and 25 subjects served as control group. Venous blood samples were taken in both groups at 4 time points: at the day of admission, the 2nd, 4th and 7th day of hospitalization. By using indirect immuno-fluorescence tests the expression of auto-antibody in the samples was semiquantitatively detected. Other biochemical indexes, such as serum amylase, urine amylase, were determined simultaneously. As well, the clinical signs or symptoms were mornitored. It was found that the expression of the auto-antibody was gradually enhanced with the development of acute pancreatitis. The inceased positive expression of auto-antibody showed a correlationship with the improvement of biochemical indexes (r=0.951) and clinical manifestations (r=0.996). There was significant difference between experimental and control groups (P<0.05). During the recovery period of acute pancreatitis, gradually increased auto-antibody expression was detectable. This antibody is against the interstitial structure of the pancreas.

To explore the effect of the extracellular ATP and its receptors on axon regeneration in the sciatic nerve defect in rats, 0.5 cm defect of the sciatic nerve was made and repaired with long arm silicon tube like a “Y” type. The single arm of the silicon tube was sutured to proximal of the sciatic nerve. 10 μl 1 mmol/L ATP in physiological saline was injected into the left chamber of the silicon tube (experimental group) and physiological saline injected into another silicon tube as a control group. In another model 1 mmol/L 10 μl ATP and 1 mmol/L 10 μl ATP+0.2 mg/ml suramin were injected respectively into two arms of the silicon tube. After 4 and 8 weeks the specimens were obtained from the silicon tube for examining the axon regeneration histologically and image analysis. All the regeneration axons grew into the silicon tube containing the ATP, but there was no axon regeneration in the silicon tube containing the ATP+Suramin or physiological saline. It was demon-strated that extracellular ATP had a powerful attraction to the regenerated axon of peripheral nerve. The suramin inhibited the axon induction of the extracellular ATP completely via the ATP receptors.

To study the relationship between polymorphism of vitamin D receptor (VDR) allele with formation of calcium oxalate calculus and find the predisposing genes of calcium oxalate calculus, we screened out 150 patients who suffered from calcium oxalate calculus. 36 of them had idiopathic hypercalciuria according to analysis of calculus component and assay of urine calcium. The polymorphisms of VDR gene Taq1, Apa1 and Fok1 were detected using PCR-RFLP technique and the correlation were analyzed between the polymorphism and urinary calculus or between the polymorphism and hypercalciuria. The difference in each genotypic frequency of the allele of promoter Fok1 between calculus group and healthy group or between idiopathic hypercalciuria calculus group and health group was significant. The content of 24-h urine calcium of those who had genotype ff was obviously higher than that of those who have other genotypes in the same group. There was no significant difference in the polymorphism of gene Apa1 and Taq1 between each two groups. It is concluded that hypercalciuria and calcium oxalate calculus were related to the polymorphism of VDR gene’s promoter Fok1 allele, but it had nothing to do with the polymorphism of gene Apa1 and Taq1. The genotype ff was a candidate heredity marker of calcium calculus disease.

In order to investigate the changes of serum hyaluronic acid (HA) and laminin (LN) levels and their clinical implication in the patients with bladder tumors, the serum HA and LN levels in 34 patients with bladder tumor and 30 cases of control group were detected by radioimmunoassay before and after operation. The results showed that the serum HA and LN levels in the patients with bladder tumors were significantly higher than those in control group (P<0.01) before operation, and decreased significantly after operation (P<0.05). The serum levels of HA and LN in infiltration tumors were higher than those in superficial tumors (P<0.05). The serum HA and LN levels in patients with lymph node metastasis were higher than those without lymph node metastasis (P<0.01). The investigation revealed that HA and LN might be involved in the malignant biology behavior of bladder tumors and could be used as important markers of assistant diagnosis and condition monitoring.

To establish a specific and sensitive radioimmunoassay (RIA) for antiarrhythmic peptide (AAP) and study serum AAP levels in normal Sprague-Dawley rats (NR) and spontaneous hypertension rats (SHR), AAP-bovine serum albumin (BSA) conjugate was prepared by ammonium bicarbonate method. The New Zealand rabbits were immunized by administering intradermally the conjugate. Then the rabbit anti-AAP serum was produced and iodinated AAP was made by Bolton-Hunter method. The RIA for serum AAP was set up and serum AAP levels in NR and SHR were determined. The minimal detectable range of the AAP RIA was (0.45±0.06) μg/L. The affinity constant for antiserum was 1.05×10⁹ L/mol, and the rate of cross-reactivity with atrial peptide (AP) and human growth hormone (hGH) were 0.12% and 0.20%, respectively. The mean recovery rate of high, medium and low doses was 97.6%, and the mean coefficients of variation for intra-and interbath-assay were (6.43±0.85)% and (9.62±1.04)%, respectively. The mean levels of AAP in NR with different age (3 months, 8–10 months and 18–20 months) were (1.75±0.13) μg/L, (1.74±0.11) μg/L and (1.79±0.15) μg/L, respectively, while those in SHR with different age (3 months, 8–10 months and 18–20 months) were (2.38±0.35) μg/L, (2.54±0.25) μg/L and (2.83±0.21) μg/L, respectively. The levels of serum AAP showed a positive correlation with blood pressure (r=0.8667,P<0.05). It was indicated that this AAP RIA had high specificity, high accuracy and good reproducibility. The levels of serum AAP had a close relation with blood pressure.

In order to investigate the role of placental isoferritin (PLF) in pathogenesis of preeclampsia and/or intrauterine growth retardation (IUGR) and its earlier predictive value, a prospective double-blinded study was performed. In 120 initial normal pregnant women at earlier third trimester (from 24 to 34 weeks), plasma placental isoferritin and nitric oxide (NO) metabolites (nitrite/nitrate) (NO₂⁻/NO₃⁻) were examined by using ELISA and Criess assay respectively. The out-come of pregnancies and birth weight of their infants were followed up. The receiver operating characteristic curves (ROC) and predictive values of PLF predicting the outcome of pregnancy with IUGR, pre-eclampsia were analyzed. Results showed that in 120 initial normal pregnant women, IUGR occurred in 15 pregnant women (IUGR group) and pre-eclampsia in 19 (pre-eclampsia group), and the remaining 86 had normal pregnancy (normal group). The levels of plasma placental isoferritin were significantly decreased in IUGR group (260.01±58.95) μg/ml and pre-eclampsia group (285.31±53.73) μg/ml as compared with those in normal group (775.62±89.32) μg/ml at earlier third trimester (bothP<0.01). The levels of plasma NO were significantly increased in IUGR group (61.57±46.22) μmol/L and pre-eclampsia group (58.37±30.52) μmol/L as compared with those in the normal group (35.29±24.46) μmol/L (bothP<0.01). There was no significant difference in plasma placental isoferritin and NO levels between IUGR group and pre-eclampsic group (bothP>0.05). The plasma placental isoferritin was negatively correlated with NO levels (r=0.329,P<0.01). The areas under ROC of PLF predicting IUGR and pre-eclampsia were 0.977 and 0.905 respectively. At the cut point of 400 μg/ml PLF level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index of PLF levels predicting the outcome of pregnancy with pre-eclampsia were 100%, 85.15%, 55.88%, 100% and 0.645 respectively. At the cut point of 390 μg/ml PLF level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index of PLF levels predicting the outcome of pregnancy with IUGR were 100%, 81.9%, 44.12%, 100% and 0.663 respectively. It was concluded that the decrease of plasma placental isoferritin levels at earlier third trimester was associated with IUGR and/or pre-eclampsia, and the endothelial cell damage may be one of its mechanisms. The plasma PLF level can be used as an earlier predictor for screening of IUGR and/or pre-eclampsia.

In order to investigate the relationship between serum testosterone level and expression of androgen receptors in ovary in relation to insulin resistance in polycystic ovary syndrome (PCOS). Serum testosterone levels were determined by radioimmunoassay in 17 patients with PCOS and 20 cases as control group. The expression of androgen receptor in ovary was detected by immunohistochemistry method. The results showed that serum testosterone level [(3.1±1.5) nmol/L] and insulin resistance index (0.85±0.49) in patients with PCOS were significantly higher than in control group (P<0.05), and showed a positive relation (r=0.65,P<0.01). The expression levels of androgen receptor in ovary of patients with PCOS were significantly higher than that in control group (P<0.05). The optical density value was positively related with insulin resistance index (r=0.59,P<0.01). It was concluded that androgen and androgen receptor could accelerate insulin resistance and the interaction of them might aggravate the pathophysiological change in PCOS.

In order to identify the distribution of gene types of β-thalassemia and reduce the birthrates of β-thalassemia major in Guiyang area, 1054 pregnant women and their spouses from Affiliated Hospital, Guiyang Medical College were screened. The positive samples were analyzed with polymerase chain reaction and reverse dot blot method (PCR-RDB). When both partners were heterozygous identified as carriers for β- thalassemia, the risk of having a fetus who was homozygous or compound heterozygous was 2.66%; the ratio of male to female was 1/1.15. Seven types of mutation were identified. CD17 and CD41–42 were dominant among them. Among the 4 cases subject to prenatal gene diagnosis, one fetus was completely normal and 3 fetuses were diagnosed as having β-thalassemia major (1 homozygous and 2 compound heterozygous). The fetuses diagnosed as β-thalassemia major were selectively terminated within two weeks. It was concluded that the birthrate of β-thalassemia major in Guiyang area was reduced and the target of improving birth outcome and child development has been achieved.

In order to study the molecular immune-pathological mechanism of spontaneous abortion (SA), immunohistochemistry techniques were used to detect the FasL expression of first trimester trophoblast in the SA patients and normal controls. High precise color-image measure system for immuno-histochemistry (HPIS) was used to determine the quantity of FasL expression. The results showed that the scale and intensity of FasL expression on the trophoblasts in SA group were significantly lower than in the control group. It is indicated that abnormal expression of FasL on trophoblasts, which damages the immunological tolerance between mother and fetus, may be one of the important mechanisms of development of SA. To induce the expression of FasL or to regulate the immunological tolerance will be a new way to treat SA.

A new method of chemiluminescent enzyme immunoassay (CLEIA) was developed and the standard curve and regression equation for determination of progesterone receptor (PR) made. The luminosity of tissue samples was tested and PR level was calculated by the regression equation. Correlation analysis revealed that there was a linear relationship between different concentrations of the standard PR samples and the corresponding values of luminosity: Y=3748+463.77X, γ=0.9958. The values of the luminosity in 38 cases of tumor tissues were determined with the highest being 267.32 fmol/mg, the lowest 3.69 fmol/mg and the mean 78.53 fmol/mg. The new method of CLEIA was a stable, creditable, specific and sensitive assay for determination of PR.

To investigate the relationship between the ATP content in human oocytes and the developmental potential of human oocytes, unfertilized oocytes from clinical IVF and immature oocytes unsuitable for ICSI were collected. The ATP content of these unfertilized andin vitro matured eggs were determined quantitatively by measuring the luminescence using an ATP-dependent bioluminescence assay. The result showed that the ATP content of unfertilized oocytes was higher thanin vitro matured ones (2.20±0.67 pmol vs 1.72±0.49 pmol,P<0.05). In unfertilized oocytes, the ATP content of those whose fertilization rates (FR) was higher than 50% was 2.43±0.60 pmol, which was significantly different from those whose FR was less than 50% (1.72±0.56 pmol), while inin vitro matured oocytes, the ATP content of those whose FR more than 50% was 1.8±0.44 pmol, slightly higher than those less than 50% (1.55±0.40 pmol), without statistical significance. There was a tendency that the ATP content of oocytes of pregnant patients was higher than those of controls, but the sample number was too small to show any significance in statistics. Briefly, there was positive correlation between the ATP content in oocytes and developmental potential of oocytes.

In order to search for a more reliable method of sorting fetal nucleated red blood cells (NRBCs) and DNA from maternal peripheral blood and to identify origin of NRBCs and DNA, NRBCs were isolated from peripheral blood of 88 pregnant women by density gradient centrifugation and fluorescence activated cell sorter (FACS) respectively. Nested polymerase chain reaction was used to detect normal male SRY gene from blood plasma DNA of 65 pregnant women. The results revealed that fetal NRBCs were found in 14 of 27 maternal samples by density gradient centrifugation. The number of cells was from 1 to 10. Using FACS, CD71⁺ cells were identified among all 61 samples. The frequency was (0.35±0.25)×10⁻²; The detectable rate of the SRY gene of blood plasma DNA from 46 women carrying male fetuses was 65.22% (30/46). Non-detectable rate for 19 women carrying female fetuses was 94.74% (18/19). It was concluded that the methods of sorting fetal NRBSs and DNA have already made great progress. The methods for fetal NRBCs and plasma DNA from maternal peripheral blood to diagnose genetic diseases seem to be the best methods of noninvasive prenatal diagnosis.

MDM2 was transfected to acute lymphoblastic leukemia (ALL) line EU-4 cell which lacks P53 expression and expresses very low levels of MDM2. The results showed that MDM2 upregulated P65 expression in mRNA level and protein level. The effect of adriamycin (ADM) on MDM2-transfected EU-4 cell was detected by MTT assay. It was found that MDM2 transfection could increase drug resistance of EU-4 cells to ADM as compared with parent cells. Since the expression of E-selectin is P65 dependent, E-selectin promoter-CAT construct and P65 and MDM2 expression plasmids were co-transfected to EU-4 cells, revealing that MDM2 increased P65-mediated transactivation of E-selectin promoter. In the absence of P65, MDM2 had no effect on the transactivation of E-selectin. Moreover, MDM2 antisense couldn’t change the transactivation of E-selectin. It was concluded that MDM2 could up-regulate transcriptionally P65 expression; MDM2 increased drug resistance of leukemia cells to ADM; MDM2 elevated NF-kappaB activity in a P53-independent manner.

To investigate whether treatment with retinoic acid (RA) could improve level of lung alveolarization and influence lung collagen in newborn rats exposed to hyperoxia, newborn Sprague-Dawley rats aged 2 days were randomly assigned to 8 groups: (1) air, (2) O₂, (3) air+NS, (4) O₂+NS, (5) air+dex, (6) O₂+dex, (7) air+RA and (8) O₂+RA. Group 2, 4 6 and 8 were kept in chambers containing 85% oxygen, the values were checked 3 times a day. The other 4 groups were exposed to room air. Level of alveolarization and lung collagen were analyzed at age of 14 or 21 days through radial alveolar counts, alveolar airspace measurements, type I, II collagen immunohistochemical methods (SP method) and image processing system. Transforming growth factor-β receptors and procollagen mRNA accumulation were examined at age of 14 days through immunohistochemical methods andin situ hybridization. Our results showed that radial alveolar counts were increased and distal airspace was enlarged in group 8. Type I collagen was markedly increased, and transforming growth factor-β receptors and procollagen mRNA were decreased by retinoic acid in bronchial epithelial cells, alveolar epithelial cells and alveolar intersitium. It is concluded that retinoic acid can partially reverse lung development arrest during exposure to hyperoxia by increasing lung collagen.

Objective Eine Datenbank der Normwerte der Hufigkeitsquoten der Osteophyten an den Lendenwirbelkörpern aufzubauen. Methoden 633 Röntgen-Übersichtsaufnahmen von männlichen Personen im Alter zwischen 20 und 87 Jahren und 607 Röntgen-Übersichtsaufnahmen von weiblichen Personen im Alter zwischen 20 und 92 Jahren ausgewertet. Eegebnisse Die Häufigkeitsquoten der Osteophyten nahmen mit zunehmendem Lebensalter zu. In den Altersgruppen 50–59 Jahren und 60–69 Jahren lagen die Häufigkeitsquoten der Osteophyten bei Männern signifikant höher als bei Frauen (P<0.01;P<0.05). Bei Männern waren am Wirbelkörper L4 am häufigsten Osteophyten vorhanden, gefolgt vom Wirbelkörper L3; bei Frauen waren am Wirbelkörper L3 am häufigsten Osteophyten vorhanden, gefolgt vom Wirbelkörper L4. Folgerung Mit Hilfe der Datenbasis ist es möglich, die Häufigkeitsquoten der Osteophyten an den Lendenwirbelkörpern durch Vergleich mit der altersbezogenen Norm quantitativ zu bewerten.

The cricoarytenoid relationship presented with spiral computed tomography was demonstrated and the reconstruction of arytenoid dislocation was presented by using multiplanar reconstruction algorithms. Fifteen patients with arytenoid dislocation documented by fiberoptic laryngoscopy and strobovideolaryngoscopy and 10 normal persons were displayed by spiral computed tomography (CT). A making design of our own had been used to diagnose arytenoid dislocation on axial CT image. Results showed that dislocation of cricoarytenoid joint was consistently demonstrated on several of the overlapping thin axial reconstructions in each of the 15 patients, in whom asymmetry of the bilateral cricoarytenoid joints was noted on axial images. It was found that on the glottic-fissure level the basal angle on abnormal side was larger in 8 patients than that on the normal side and smaller in 7 patients in patient group, whereas right basal angle was equal to the left in 8 subjects, except 2 in control group. There was statistically significant difference in the number of the equal to two basal angles of glottic fissure between control group and patient group (P<0.025). High-quality sagittal and coronal reconstructive images often were helpful in confirming or clarifying the complex arytenoid orientations. The findings that two-side basal angle was not equal in triangle of glottic fissure can be used as an objective parameter to diagnose arytenoid dislocation. Spiral CT is a useful adjunct in the diagnosis and treatment of dislocation of cricoarytenoid joint.

The connective tissue growth factor (CTGF) expression in cultured corneal fibroblasts and its effect on corneal fibroblasts proliferationin vitro were examined. Total RNA was extracted from early passaged rabbit corneal fibroblasts by guanidine isothiocyanate one-step method and mRNA was reversely transcripted into complementary DNA (cDNA). Specific CTGF primers were used in the PCR reaction and the products were analyzed by electrophoresis to determine the expression of mRNA for CTGF against DNA marker. House-keeping gene GAPDH was used as control. Different concentrations of CTGF (0.5, 5, 50, 500, 5 000, 50 000 ng/ml) were added into the third passaged corneal fibroblast culture system, and its effect on corneal fibroblast proliferation was measured by MTT method. The results showed that compared with a GAPDH 450 bp band, CTGF RT-PCR product showed a specific 120 bp band as expected. CTGF produced a dose-dependent increase in the proliferation of corneal fibroblasts but it inhibited fibroblast proliferation at higher concentrations (5000 and 50 000 ng/ml). It was concluded that proliferating corneal fibroblasts produce CTGF and CTGF helps to promote corneal fibroblast proliferation. The results indicated that CTGF might be involved in the corneal wound healing after photorefractive keratectomy in which corneal fibroblasts are activated to proliferate and secrete growth factors that in turn promote corneal fibroblast proliferation.

Whether cultured human trabecular meshwork cells express transforming growth factor-β₂ (TGF-β₂) messenger RNA (mRNA) and protein was investigated. Total RNA of 10⁶ cultured human trabecular meshwork cells was extracted with TRIZOL reagent, reverse transcriptase-polymerase chain reaction (RT-PCR) were used for detection of TGF-β₂ messenger RNA, and the PCR product was verified by sequencing. Immunohistochemical staining was used to detect TGF-β₂ protein. The results showed that a single RT-PCR amplified product was obtained, and the sequence was homologous to the known sequence. TGF-β₂ immunostaining was positive. It was concluded that trabecular meshwork cells could produce TGF-β₂ and contribute to the presence of TGF-β₂ in trabecular meshwork microenvironment as well as aqueous humor. Trabecular meshwork cells were affected by TGF-β₂ not only through paracrine, but also autocrine action. Whether abnormal changes in TGF-β₂ production contribute to the pathogenesis of primary open-angle glaucoma is worth further investigation.

To investigate the role of IL-18 and IL-18 binding protein (IL-18BP) in the pathogenesis of psoriasis, PT-PCR was used to semi-quantitatively analyze the mRNA expression of IL-18, AcPL (accessory protein-like) and IL-18BP in 12 psoriatic lesions and in 6 normal skin tissues. The results showed that IL-18 and AcPL mRNA were present in all specimens from 6 normal skin and 12 pieces of psoriatic skin, and IL-18 and AcPL showed relatively stronger intensity in the psoriatic skin compared with those seen in normal skin (P<0.01). Amplified bands indicating the expression of IL-18BP mRNA were weakly positive in all samples of normal skin. In contrast, the PCR products for IL-18BP were distinctly visible in all specimens of psoriatic lesions (P<0.01). These findings demonstrate that IL-18 is constitutively synthesized by human keratinocytes and involved in the development of the Th1 response in psoriatic lesions, and its bioactivity appears to be closely regulated by cutaneous inflammation.

To demonstrate the direct effects of cadmium on activities of bone morphogenetic protein (BMP), a complex containing BMP and cadmium chloride (CdCl₂) was implanted beneath the abdominal skin of young male Wistar rats. The activity of BMP was studied by observing the histological changes, and measuring the activity of alkaline phosphatase (ALP) and acid phosphatase (ACP) and calcium content of the implants at different time points. Our results showed that during bone formation induced by BMP, cadmium inhibited the activities of osteoblasts and osteoclasts, and slowed the deposition of calcium. It is concluded that cadmium can directly affect biological activities of BMP directly.

This article was designed to forecast the supply and demand of medical postgraduate from 2001 to 2010 and bring forward the development strategy of medical postgraduate education in Hubei province. The line regression, the ratio of health manpower to population, grey dynamics model (1,1) was employed to forecast the supply and demand of medical postgraduate according to the corresponding data from 1991 to 2000 in Hubei province. The results showed that the number of health professionals of Hubei province in 2010 would attain 265892, the graduates’ proportion of which would be about 1.8%; and the demand and supply of medical postgraduate would be 4699 and 2264 respectively. To improve conditions actively to attract and cultivate excellent professionals, especially the senior medical scholars, are effective measures to adjust the degree structure of health manpower as soon as possible in Hubei province.