Evaluation of renal clinicopathological changes in IgA nephropathy by urinary podocytes excretion and podocalyxin expression.

OBJECTIVE: To explore the association of urinary podocyte excretion and renal expression of podocyte-specific marker podocalyxin (PCX) with clinicopathological changes in immunoglobulin A nephropathy (IgAN). METHODS: Morning urine samples from IgAN patients and healthy controls were collected. The expression of glomerular PCX was quantified in 50 IgAN patients diagnosed by renal biopsy. IgAN was classified based on the Lee's Grading system and scored according to the Katafuchi semiquantitative criteria. Morphological evaluation of podocyte was determined by electron microscopy. RESULTS: The amount of urinary podocytes in the IgAN patients was significantly higher than that in the healthy controls (p < 0.01). Pairwise comparison among Lee's grades of IgAN showed that the median of urinary podocytes in Lee's I-II group was lower than that in Lee's III, IV, and V groups (p < 0.05); group III lower than group V (p < 0.05). The positive rate of urinary podocytes was the highest in Lee's IV and V groups (100%), and lowest in Lee's I-II group (55%). Multiple comparison among groups of Lee's grades of IgAN showed that the glomerular PCX expression in Lee's I-II group was higher than that in Lee's III, IV, and V groups (p < 0.05); groups III and IV higher than group V (p < 0.05). The amount of urinary podocytes in IgAN patients was negatively correlated with PCX expression (r = -0.702, p < 0.01), but positively correlated with 24-h urinary protein (r = 0.465, p < 0.01) and glomerular (r = 0.233, p < 0.01) and renal tubular pathological scores (r = 0.307, p < 0.05). The glomerular PCX expression was negatively correlated with 24-h urinary protein (r = -0.367, p < 0.05) and glomerular (r = -0.560, p < 0.05) and tubular pathological scores (r = -0.377, p < 0.05). Electron microscopy showed significant changes in podocytes of IgAN, especially in the foot process. CONCLUSION: The amount of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be a marker of IgAN progression.

Molecular characterization of a Chinese pedigree with beta-thalassemia intermedia.

Hereditary persistence of fetal hemoglobin (HPFH), often associated with mutations in the beta-globin gene cluster, is normally benign, but a person carrying both HPFH and another beta-thalassemia (beta-thal) mutation will develop serious anemia. These people might be erroneously diagnosed as having homozygous beta-thal with common reverse dot-blot methods. Here we report a 5-year old boy with thalassemia intermedia, who is a compound heterozygote for the rare HPFH-6 deletion with codons 41/42 (-TCTT) beta(0)-thal, who inherited the deletion from his mother and the beta(41/42) mutation from his father.

[Chronic toxicity of 97% isopropyl thioxanthone in rat by oral administration for 2 years].

OBJECTIVE: To study the oral chronic toxicity of 97% isopropyl thioxanthone (97% ITX) in rats, determine the no-observed adverse effect levels (NOAEL). METHODS: Four groups of rats were fed with foodstuff containing 97% ITX in the dosage of 1000.0, 250.0, 62.5 mg/kg respectively for 2 years. The general behavior, body weight, food availability ect. were observed during the experiment. At the end of the experiment, blood and urine samples were collected for routine and biochemical assays. The internal organs were taken for calculating their organ coefficients and histopathological examinations. RESULTS: During the experimental period, no obvious abnormality were found in the experimental animals. The body weight and the total food availability rate in the high dosage group of male were lower than that of control (P < 0.05). Hematology examination showed that the quantity of Hb and RBC in high dosage groups of both the male and female and Hb in the male middle group were all lower than the control group (P < 0.01 or P < 0.05). Analysis of correlation indicated that r = -0.433, P < 0.01 in male, r = -0.337, P < 0.01 in female of Hb; r = -0.266, P < 0.05 in male, r = -0.317, P < 0.01 in female of RBC. There were obviously negative correlation. Serum biochemistry examination showed the concentration of CHO in the high and middle dosage treated rats of male and female were higher than that of the control (P < 0.01 or P < 0.05). Analysis of correlation indicated that r = 0.497, P < 0.01 in male, r = 0.417, P < 0.01 in female. No abnormality were found in urine examination. The organ weight and organ coefficient such as liver, were higher than control group (P < 0.01). The result of histopathological examinations displayed that the renal tubule Cast and the tubulointerstitial nephritis in the treated groups were higher than that of control group (P < 0.01). CONCLUSION: 97% ITX could obviously interfere with the animals' physical condition, and reduce the number of RBC and the concentration of Hb in the blood, interact metabolism of lipoid and induce the concentration of CHO in the serum. The livers of the treated rats are compensatory enlarged. And kidneys of the poisoning animals are damaged. The 2 years oral NOAEL of 97% ITX in rats are more than 4.63 mg/kg for female rats, and larger than 4.06 mg/kg for male rats.

[Nephrotoxicity of high- and low-osmolar contrast media: Protective role of fosinopril or telmisartan in a rat model].

OBJECTIVE: To compare the nephrotoxicity of high- and low-osmolar contrast media (HOCM and LOCM), and to determine the protective role of fosinopril or telmisartan and its possible mechanism. METHODS: Forty eight healthy SD rats were randomly divided into 6 groups: a normal control group, a glycerol control group, a low-osmolar contrast media (LOCM) group, a high-osmolar contrast media (HOCM) group, a fosinopril group, and a telmisartan group. Glycerine for inducing kidney damage was given to all rats except the normal control group. Twenty-four hours after the injection of glycerine, the mixed fosinopril suspension (10mg/kg) or telmisartan (5mg/kg) was poured into the stomach in the preventive group. Serum creatinine (SCr) and plasma angiotensin II (AngII) levels were detected by an automatical biochemical analyzer and radioimmunoassay; caspase-3 activity and claudin-1 expression of the renal tissue were detected by fluorometric method and immunohistochemical method. The renal injury was assessed by hematoxylin and eosin (HE) staining and terminal deoxynucleotide mediated nick and labeling (TUNEL) staining, respectively. RESULTS: In diatrizoate-injected rats, SCr and AngII levels were increased (P<0.05). Expression of claudin-1 protein and caspase-3 activity in the renal tissue was upregulated. The histologic changes and percentage of apoptotic cells were milder in the LOCM rats than those in the HOCM rats. In the group pretreated with fosinopril or telmisartan, no increase in the levels of SCr and AngII was discovered. The expression of claudin-1 protein and caspase-3 activity was significantly lower than that in the HOCM group. The renal injuries induced by diatrizoate were alleviated. CONCLUSION: Both HOCM and LOCM could cause cellular apoptosis in the kidney.LOCM was less toxic to rat kidney than HOCM. Nephrotoxicity induced by HOCM might be related to caspase-3, claudin-1 and AngII. Fosinopril or telmisartan may protect the renal tissue from nephrotoxicity induced by diatrizoate.

[Effects of Alternanthera philoxeroides Griseb against dengue virus in vitro].

OBJECTIVE: To investigate the effects of Alternanthera philoxeroides Griseb extracts against dengue virus in vitro. METHODS: MTT assay and observation of cytopathic effect (CPE) were carried out to determine the cytotoxicity of Alternanthera philoxeroides Griseb on C6/36 cell lines and its effects on dengue virus. RESULTS: None of the 4 kinds of Alternanthera philoxeroides Griseb extracts exhibited obvious cytotoxicity on the cells at different concentrations with the exception of that over 320 microg/ml. The 4 extracts all showed inhibitory effects on dengue virus. Statistical analysis of TD(50) and ED(50) by Probit regression method suggested that extracts from coumarin had the lowest toxicity on the cells (TD(50)=535.91), whereas extracts from petroleum ether showed the strongest inhibitory effects on dengue virus (ED(50)=47.43) among the 4 extracts. CONCLUSION: Alternanthera philoxeroides Griseb possesses antiviral effects on dengue virus in vitro.

Detection of serum Chlamydia pneumoniae (Cpn) immune complex and Cpn antibody in patients with coronary heart disease.

OBJECTIVE: To understand the state of Chlamydia pneumoniae (Cpn) infection in patients with coronary heart disease (CHD), and explore the relationship between Cpn infection and the gonesis and progressin of CHD. METHOD: By adopting sandwich method with double antibodies and indirect ELISA method, we detected serum Cpn immune complex and IgG antibody in 160 CHD patients (confirmed by coronary artery angiography) and 40 subjects without CHD. RESULTS: The positivity rate of Cpn IgG type immune complex in the CHD group (63.8%) was significantly higher than that in the control group (40.0%, P<0.05), similar to the results of Cpn IgG antibody detection (65.0% vs 32.5%, P<0.05). CONCLUSION: Cpn infection is closely connected with the CHD, but their exact relationship needs further exploration and study.

[Establishment of a rat model of mycoplasma pneumonia and the treatment].

OBJECTIVE: To establish a rat model of mycoplasma pneumonia (MP) for investigating the pathogenesis of MP and its therapy with drugs. METHODS: Thirty Wistar rats were randomly divided into 6 groups (n=5), including a control group, a MP model group, a erythromycin lactobionate group and 3 erythromycin microspheres groups (high, middle, and low dose groups). With the exception of those in the control group, all the rats received intranasal MP administration followed by corresponding treatments administered via tail vein injection. At different time points after inoculation of the pathogen, the lungs of the rats were taken for histopathological scoring. RESULTS: In the MP model group, the lung pathology was characterized by patchy interstitial pneumonitis with predominantly lymphocyte infiltration and mucosal edema. The bronchiolar walls became thickened and the lumens narrowed. In erythromycin lactobionate and erythromycin microspheres treatment (high and middle dose) groups, clear cell boundaries were observed in the lungs where no obvious pathological changes were found. RT-PCR amplification showed positive results of MP RNA in the model group, erythromycin lactobionate group and erythromycin microsphere groups. CONCLUSION: The approach described is practicable to establish rat models of MP. Erythromycin microspheres can effectively relieve the lung inflammations and has therapeutic effect on MP.

[Antimicrobial effects of erythromycin microspheres against Mycoplasma Pneumoniae in rats].

OBJECTIVE: To evaluate the antimicrobial effects of erythromycin microspheres against Mycoplasma pneumoniae in rats. METHODS: With erythromycin lactobionate as the positive control, erythromycin microspheres at 3 non-toxic doses (0.1, 0.5, and 1.2 g.kg(-1).d(-1)) were administered intragastrically for 6 consecutive days in Wistar rats with Mycoplasma pneumoniae infection. The general condition and lung index of the rats were observed and measured to assess the therapeutic effects of the treatments against Mycoplasma pneumoniae infection. RESULTS: The erythromycin microspheres at 0.1, 0.5, 1.2 g.kg(-1).d(-1) significantly alleviated the symptoms of the rats infected with Mycoplasma pneumoniae and reduced the pulmonary index of the infected rats from 1.75 to 1.45, 1.38 and 1.25, respectively (P < 0.01). An obvious dosage-effect relationship was noted between the dose of erythromycin microsphere and the tissue pathologies due to the infection. CONCLUSION: Erythromycin microspheres possess strong activity against Mycoplasma pneumoniae in rats.

[Rapid detection of oxacillin and erythromycin resistance genes in Staphylococcus aureus using multiplex PCR].

OBJECTIVE: To establish a rapid multiplex PCR (MPCR) detection system of oxacillin and erythromycin resistance genes in Staphylococcus aureus (S. aureus) and evaluate the genotype distribution of the genes associated to mecA, ermA and ermC resistance in Guangzhou. METHODS: The S. aureus strains were identified and susceptibility tests were performed using VITEK-60 or PHOENIX-100 system. The inducible resistance to clindamycin of strains with of erythromycin resistance was conducted using D-test, and the MPCR system of for detecting the antibiotic resistance genes was optimized. RESULTS: The MPCR assay for detecting the resistance genes was constructed successfully. According to the results of MPCR, the positivity rates for mecA, ermA and ermC genes among the 124 strains of S. aureus isolated from clinical samples were 56.5%, 50% and 33.9%, respectively. Good correlation was observed between the antibiotic resistance phenotypes and the S. aureus genotypes. mecA were detected in all the methicillin-resistant S. aureus strains, and ermA and/or ermC in 97.7% of the S. aureus strains with erythromycin resistance. CONCLUSION: This MPCR system allows rapid and reliable analysis of antibiotic resistance genotypes of S. aureus isolated from clinical samples. mecA, ermA, and ermC genes are among the predominant genetic determinants for the resistance to oxacillin and erythromycin in S. aureus isolates in Guangzhou.

[Effects of Alternanthera philoxeroides Griseb against respiratory syncytial virus infection in mice].

OBJECTIVE: To investigate the effect of an oral preparation of Alternathera philoxeroides Griseb (APG) against respiratory syncytical virus (RSV) in mice. METHODS: APG preparation was administered orally in RSV-infected mice at different daily doses (2.5, 4.5 and 6.5 g/kg) to observe the therapeutic effect of the preparation. RESULTS: Distinct differences were observed between the death rate of the mice treated with APG at daily dose of 4.5 and 6.5 g/kg and that of the untreated mice with infection. After AGP treatment of the mice at 6.5 g/kg, the detection rate of the virus was 31.3% in the blood and 37.5% in the lung tissue, significantly lower than that in the untreated mice. The virus detection rate was 43.8% in the lung tissues of mice treated with APG at 4.5 g/kg, also significantly lower than that in the untreated control. APG treatment at the 3 doses resulted in different lung indices from that of the control. CONCLUSION: APG may be effective for treatment of RSV infection.

[Identification and tissue localization of intermediate filament protein in Angiostrongylus cantonensis].

OBJECTIVE: To identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization. METHODS: Recombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis. RESULTS: The antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma. CONCLUSION: The antigen IF is distributed in the intestine wall of A. cantonensis.

[Induced differentiation and signaling factor PTEN expression of 3T3-L1 adipocytes].

OBJECTIVE: To optimize the condition for inducing the differentiation of 3T3-L1 preadipocytes into adipocytes and study the expression of PTEN tumor suppression gene in this process, aiming to understand the regulatory role of PTEN in normal adipocyte differentiation and collect laboratory evidence for developing drugs targeting PTEN. METHODS: The differentiation of 3T3-L1 preadipocytes cultured in high-glucose DMEM were induced according to 2 protocols with different combinations of dexamethasone, isobutylmethylxanthine (IBMX) and insulin, and the resultant adipocytes were identified by oil red O staining. The total proteins of 3T3-L1 were extracted and analyzed by Western blotting, and PTEN homology between mice and human was analyzed by bioinformatic method. RESULTS: For optimized 3T3-L1 differentiation, 3T3-L1 cells were initially induced with the combination of 1 micromol/L dexamethasone, 0.5 mmol/L IBMX and 5 microg/ml insulin for 48 h, followed by treatment with 5 microg/ml insulin in 4.5 g/L glucose DMEM for 48 h, which resulted in high differentiation rate of 3T3-L1 cells (up to 90% on the 10th day) with unified morphology and size. PTEN expression varied quantitatively in the process of differentiation, especially low on the 12th day as compared with those measured on days 4, 6 and 9. The mice PTEN mRNA shared 96% homology and PTEN amino acid 100% homology with their human counterparts. CONCLUSION: Endogenous PTEN expression is down-regulated during 3T3-L1 differentiation, suggesting that PTEN may enhance insulin sensitivity and promote adipogenesis under physiological conditions.

[Investigation of viral pathogens contributing to habitual abortion].

OBJECTIVE: To investigate the association of Coxsackie B virus (CBV) with habitual abortion. METHODS: CBV IgM antibody, viral RNA and virions were detected in 86 women with habitual abortion and 40 with induced abortion by means of enzyme-linked immunosorbent assay (ELISA), RT-PCR and virus isolation, respectively. RESULTS: The positivity rate of CBV IgM were 87.2% and 35% in the two groups, respectively, and the detection rate of the viral RNA was 53.5% and 17.5% in blood lymphocytes, and 59.3% and 17.5% in the placentas. The virions were found in the placentas in 41.9% and 6.9% of the women, respectively. The positivity rates of CBV IgM, viral DNA and virions showed significant difference between the two groups (P<0. 01). CONCLUSION: CBV might be one of the causes responsible for habitual abortion.

[Relationship between cyclins and prognosis of acute leukemia].

BACKGROUND & OBJECTIVE: Cell proliferation and differentiation are directed by cell cycle mechanism. When tumor cells proliferate abnormally, cyclins, which are positive agents of cell cycle, may be expressed abnormally at the same time. Now many references proved that cyclins are highly expressed in solid tumors. This study was designed to investigate the relationship between expression of cyclins and prognosis of acute leukemia. METHODS: Sixty-eight cases of acute leukemia were enrolled. Reverse transcription polymerase chain reaction (RT-PCR) was performed on tumor samples to examine the expression of cyclin A, cyclin D, cyclin E. All samples were divided into three groups: acute leukemia in complete remission (12 cases), newly diagnosed acute leukemia (16 cases) and refractory leukemia (40 case). Samples of benign hemopoietic patients were used as normal control (15 cases). RESULTS: All the 15 cases in the control group were negative of cyclin mRNA. No difference of cyclin A mRNA expression was shown between control group and the three experiment groups (P >0.05). But expression of cyclin D and cyclin E mRNA was significantly different between them (P< 0.01). There was no difference of expression of cyclin D and cyclin E mRNA among CR patients with acute leukemia(P >0.05), while the expression of cyclin D mRNA is significantly higher than that of cyclin E in refractory leukemia group. Furthermore the expression of cyclin D mRNA in recurrent refractory leukemia patients is significantly higher than that newly diagnosed cases (P< 0.05). All cyclins mRNA positive cases were divided into single cyclin gene expressed group and multiple cyclins gene expressed group. The positive rates of them were counted. No difference was found between complete remission group and refractory leukemia group (P >0.05). CONCLUSION: Cyclin D may act as a prognostic marker for acute leukemia. The amount of cyclins expressed cannot be used as a prognostic factor for acute leukemia.