Association of the LMNA gene single nucleotide polymorphism rs4641 with bdilated cardiomyopathy.

Recently, studies on the pathogenesis of dilated cardiomyopathy (DCM) have focused on the underlying molecular biology and the association between single nucleotide polymorphisms (SNPs) and disease. This study was designed to explore the association between the rs4641 SNP of the LMNA gene and DCM in order to identify a new gene locus related to DCM. Polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing were employed to detect and genotype rs4641 in 198 patients with DCM and 160 healthy controls. Genotype and allele frequencies were compared to discover their relationship and logistic regression was used to assess the risk of DCM associated with the polymorphic variants. In the DCM group, the frequencies of the TC and TT genotypes and the T allele of rs4641 were remarkably higher than those in the control group (P < 0.01). According to risk analysis, taking the CC genotype as a reference, both the TC and TT genotypes increased the risk of DCM pathogenesis, with OR (95%CI) values of 5.957 (2.903- 12.222) and 6.424 (2.156-19.141), respectively. Taking the C allele as the reference, presence of the T allele was found to increase DCM risk, with OR (95%CI) of 5.295 (3.121-8.983). These results suggested that the C to T mutation at the rs4641 locus of LMNA could enhance the risk of DCM, and that rs4641 represented a genetic susceptibility locus. Therefore, it was concluded that the LMNA rs4641 SNP was associated with DCM risk, which indicated that LMNA is a susceptibility gene for DCM.

Location and characterization of GAPDS in male reproduction.

Sperm-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDS) originates from spermatogenic cells of the human testis. A strong staining of GAPDS was detected in epididymal epithelium, especially in principal cells and basal cells of the epithelium. GAPDS also bound to the fibrous sheet of the sperm tail and inhibited the motility and penetration ability of sperms. The rat model showed that at postnatal day 28 the spermatogenic cells began to express GAPDS protein. By day 60 its expression decreased in spermatogenic cells while it increased in Sertoli cells. After sexual maturation (120 days) GAPDS protein was expressed in both Sertoli cells and elongated sperms. The expression of GAPDS gradually increased with age in the epididymis.

The application of chromosome abnormality chip detection in male infertility.

OBJECTIVE: To discuss the application of microarray technology in the diagnosis of male infertility. METHODS: Sixteen loci, including a sex-determining region on the Y chromosome, were investigated by polymerase chain reaction (PCR) in infertile male patients. Chromosome abnormality chip with 180 000 probes was used to detect small deletion, small amplification and loss of heterozygosity. RESULTS: By PCR, nine of 103 infertile patients were found to have sequence-tagged site microdeletions. Microdeletions were not observed in control samples. The deletions detected by PCR were present in six azoospermic men (6/44, 13.6%) and in three oligoasthenoteratozoospermic (OATS) men (3/59, 5%).The overall frequency of microdeletions in infertile men was 8.7% (9/103). Chromosome abnormality chip detection 500+ detected more amplification or deletion in 51 infertile patients and the overall frequency of microdeletions in infertile men was 49.5% (51/103). CONCLUSION: Chromosome abnormality chip detection system provides a sensitive, economic and high-throughput method for detecting the deletion or amplification of genomic DNA sequences of infertile patients. Not only can it identify Yq deletions, but it can also find other chromosome abnormalities and facilitate the understanding of male infertility.

MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5.

The article "MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5, by C.-M. Sun, G.-M. Zhang, H.-N. Qian, S.-J. Cheng, M. Wang, M. Liu, D. Li, published in Eur Rev Med Pharmacol Sci 2019; 23 (15): 6436-6444-PMID: 31378882" has been withdrawn from the authors due to some inaccuracies (some data cannot be repeated by our further research). The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18525.

[Relationship of telomere length, mitochondrial DNA copy number of peripheral blood with hypertension in coal miners].

Objective: To explore the relationship of telomere length, mitochondrial DNA copy number of peripheral blood with hypertension and the interaction between telomere length and mtDNA-CN on hypertension in coal miners. Methods: A case control study was conducted in a coal mine of Shanxi province from July to December of 2013, in which 325 healthy workers were selected as the control group and 378 workers with hypertension as the case group. The information about general demographic characteristics and life behavior habits of the subjects were collected through questionnaire. Levels of telomere length and mtDNA-CN in peripheral blood were detected by real-time PCR. Unconditional logistic regression was used to examine the association between hypertension and telomere length, mtDNA-CN. The interaction test between telomere length and mtDNA-CN on hypertension was performed by adding the interaction term in the corresponding model. Results: The mean telomere length of the workers in the case group was (1.50±0.55) kb, and that of the control group was (2.01±0.62) kb, the difference between two groups was significant (t=11.68, P<0.001). The correlation analysis showed that telomere length was positively correlated with mtDNA-CN (r=0.157, P=0.002) in the case group. Multivariate analysis showed that telomere length (OR=4.408, 95%CI: 3.012-6.452), age (OR=0.417, 95%CI: 0.284-0.613), BMI (OR=1.357, 95%CI: 1.162-1.584), monthly household income level (OR=0.656, 95%CI: 0.553-0.778) and work duration (OR=1.249, 95%CI: 1.100-1.417) were influencing factors of hypertension. The multiply interaction between telomere length and mtDNA-CN was significant on hypertension (OR=1.267, 95%CI: 1.094-1.468). Conclusions: The results suggest shorter telomere length is a risk factor of hypertension. There is a multiply interaction between telomere length and mtDNA-CN on hypertension. However, the association between mtDNA-CN and hypertension was not found.

MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5.

OBJECTIVE: To elucidate the correlation between microRNA-1266 (miR-1266) and prostate cancer (PCa) progression, and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression level of miR-1266 and protein arginine methyltransferase 5 (PRMT5) in PCa tissues and cell lines was first detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). After up-regulating or down-regulating miR-1266 expression in cells, cell proliferation, migration and invasion abilities were detected. Possible target genes of miR-1266 were predicted and validated by bioinformatics analysis and dual-luciferase reporter gene assay, respectively. Finally, abnormal expression of PRMT5 was ascertained after transfection. RESULTS: MiR-1266 was lowly expressed in PCa tissues and cell lines, whereas PRMT5 exhibited the opposite results. Up-regulated expression of miR-1266 significantly inhibited the proliferation, migration and invasion abilities of PC-3 cells. However, the growth and migration of DU145 cells with low miR-1266 expression were significantly accelerated. Meanwhile, the number of invading cells was significantly increased. PRMT5 was verified as a potential target gene of miR-1266. Furthermore, results found that miR-1266 was negatively correlated with PRMT5. In addition, the expression of PRMT5 was remarkably decreased after miR-1266 overexpression, which could be restored after knockdown of miR-1266. CONCLUSIONS: MiR-1266 inhibits the growth and metastasis of PCa by targeting PRMT5. We may provide a potential and prospective therapeutic target for PCa.

Increased regional cerebral perfusion in contralateral motor and somatosensory areas after median nerve stimulation therapy.

OBJECTIVE: To evaluate change in regional cerebral perfusion (rCBF) after median nerve stimulation (MNS) therapy in brain-damaged patients. METHODS: Twelve brain-damaged patients received 12 courses of MNS. Technetium-99m-ethyl cysteinate diethylester (99mTc-ECD) SPECT was performed before and 4 weeks after MNS initiation. Clinical response was assessed by Glasglow coma scale or clinical improvement. 12 MNS patients were grouped as good responder (GR) (n = 6) and poor responder (PR) (n = 6) according to therapy response. Scan images were analyzed by Statistical Parametric Mapping 2 (SPM2). RESULTS: In the GR group, paired Student t test between the pre- and post-MNS images showed 2 activation clusters over the left frontal and parietal lobes, including regions of the precentral gyrus, middle frontal gyrus, superior frontal gyrus, subgyral, inferior parietal lobule, and postcentral gyms (corresponding to Brodmann areas 4, 6, and 40). In the PR group, paired Student t test did not show any activation clusters. Clusters with significant differences between the GR and PR groups shared no mutual voxels with those clusters having significant regional effects after MNS in the GR group. CONCLUSIONS: Median nerve stimulation enhanced the rCBF of the contralateral motor and somatosensory cortex, which is compatible with the few previous studies using other modalities.

BAY-11-7082 induces apoptosis of multiple myeloma U266 cells through inhibiting NF-κB pathway.

OBJECTIVE: To study the effects of BAY-11-7082 on proliferation and apoptosis of U266 cells and its mechanism of action. MATERIALS AND METHODS: Multiple myelomas U266 cells were cultured and divided into control group and gradient-concentration BAY-11-7082 groups (1 µmol/L, 2 µmol/L, 4 µmol/L and 8 µmol/L). Cells in BAY-11-7082 groups were treated with drugs in different concentrations for 4 h, while those in control group were added with an equal volume of solvent. The cell viability was detected via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and lactate dehydrogenase (LDH) release assay was used to detect the cytotoxicity. Furthermore, cells in low-concentration and high-concentration BAY-11-7082 groups were compared with those in control group. The cell proliferation level was evaluated via cell cycle assay, the interleukin-6 (IL-6) level in cells was detected via enzyme-linked immunosorbent assay (ELISA), and the ß-catenin protein expression was detected via Western blotting. Moreover, flow cytometry and Hoechst staining were performed to detect the number of apoptotic cells, and the apoptosis level was detected via caspase3 activity and apoptosis-related protein expression. Finally, the levels of p65, p50 and inhibitor kappa B kinase ß (IKKß) were detected via polymerase chain reaction (PCR), and the expressions and changes of phosphorylated (p)-p65 and p-IKKß were detected via Western blotting. RESULTS: BAY-11-7082 could reduce the U266 cell viability and increase the cytotoxic effect. Based on gradient concentration, 2 µmol/L was selected as the low concentration, while 4 µmol/L was selected as the high concentration. Compared with those in control group, the number of cells in the S and G2/M phases in drug administration groups was significantly decreased, but that in the G0/G1 phase was significantly increased. Besides, the secretion of IL-6 in cells in drug administration groups was significantly decreased compared with that in control group. The ß-catenin protein expression was decreased in drug administration groups, and there was also a difference between high concentration group and low concentration group. Flow cytometry and Hoechst staining showed that the proportion of apoptotic cells in drug administration groups was significantly increased. Western blotting and detection of caspase3 activity revealed that the expression and activation of apoptosis-related protein were increased in drug administration groups. It was found in the detection of nuclear factor-κB (NF-κB) pathway that the NF-κB pathway was inhibited in drug administration groups, and there was also a statistically significant difference between high concentration group and low concentration group. CONCLUSIONS: BAY-11-7082 inhibits the proliferation and induces the apoptosis of U266 cells through inhibiting NF-κB pathway.

[Research on potential interaction between mitochondrial DNA copy number and related factors on risk of hypertension in coal miners].

Objective: To investigate the effects of mitochondrial DNA (mtDNA) copy number in peripheral blood and related factors on the risk of hypertension in coal miners. Methods: A case-control study was conducted in 378 coal miners with hypertension and 325 healthy coal miners recruited from Datong Coal Mine Group. A standard questionnaire was used to collect their general information, such as demographic characteristics, habits and occupational history. Fluorescence quantitative PCR was performed to detect the copy number of mtDNA. Logistic regression model was applied for identifying the related risk factors of hypertension and analyzing the interaction between mtDNA copy number and risk factors. Results: The prevalence of hypertension of high mtDNA copy number was lower than mtDNA copy numberin 0-5.67 group, but the difference was not statistically significant (P=0.414). Alcohol drinking (OR=1.80, 95% CI: 1.26-2.56), family history of hypertension (OR=1.74, 95% CI: 1.20- 2.50), work shifts (OR=0.69, 95% CI: 0.48-0.99), education level (P=0.012) and family monthly income level (P=0.001) were related to the prevalence of hypertension. There were potential interactions between mtDNA copy number and alcohol drinking, family monthly income level, family history of hypertension, respectively. Alcohol drinking was a risk factor for hypertension [1.77 (1.25-2.50)]. Potential interactions between mtDNA copy number and alcohol drinking reduced the risk of hypertension (OR=1.20, 95% CI: 1.07-1.35). Family history of hypertension was a risk factor for hypertension [1.81(1.26-2.59)]. Potential interactions between mtDNA copy number and family history of hypertension reduced the risk of hypertension (OR=1.24, 95%CI: 1.09-1.41). Family monthly income level was a protect factor for hypertension [0.55(0.46-0.66)]. Potential interactions between mtDNA copy number and family monthly income level increased the protection role of hypertension (OR=0.90, 95% CI: 0.86-0.94). Conclusion: mtDNA copy number variation was not significantly associated with the prevalence of hypertension in coal miners, but mtDNA copy number showed multiplication interaction on the prevalence of hypertension with alcohol drinking, family monthly income level as well as family history of hypertension and made their influences weaken.

Overexpression of microRNA-21 in peripheral blood mononuclear cells of patients with B-cell non-Hodgkin's lymphoma is associated with disease stage and treatment outcome.

OBJECTIVE: We wished to assess the association between microRNA-21 (miR-21) and disease stage and treatment outcome in patients with B-cell non-Hodgkin's Lymphoma (B-NHL). PATIENTS AND METHODS: A total of consecutive 128 patients with B-NHL were enrolled; 30 healthy individuals served as controls. qPCR assay was utilized to quantify expression levels of miR-21 in peripheral blood mononuclear cells (PBMC; Ficoll isolation protocol). Expression of the miR-21 target, phosphatase and tensin homolog (PTEN), was assessed by Western blot analysis. RESULTS: miR-21 was overexpressed in PBMC of patients with B-NHL (p < 0.05 vs. healthy individuals). Furthermore, miR-21 expression levels were significantly higher in patients with the stage III/IV B-NHL (p < 0.05 vs. stage I/II B-NHL). After chemotherapy, miR-21 expression levels were significantly decreased in patients in complete remission and became comparable to those of healthy individuals. Also, miR-21 expression levels were lower in patients treated with chemotherapy combined with rituximab. There was a negative association between miR-21 overexpression and post-chemotherapy survival rates of the patients. Expression of PTEN was significantly lower in patients with B-NHL (p < 0.05 vs. healthy individuals). CONCLUSIONS: Overexpression of miR-21 is associated with disease stage and treatment outcome of B-NHL. This potentially involves negative modulation of PTEN.

Syntheses and structure-activity relationships of taxoids derived from 14 beta-hydroxy-10-deacetylbaccatin III.

A series of new taxoids derived from 14 beta-hydroxy-10-deacetylbaccatin III was synthesized by means of the beta-lactam synthon method. Most of the new taxoids thus synthesized possess excellent cytotoxicity against human ovarian (A121), non-small-cell lung (A549), colon (HT-29), and breast (MCF-7) cancer cell lines, and several of these taxoids show subnanomolar IC50 values which are severalfold to 1 order of magnitude better than those of paclitaxel and docetaxel. Modifications at the 3'- and 3'-N-positions exert marked effects on the activity. For the substituents at C-3', the cytotoxicity decreases in the order 2-furyl approximately 2-methyl-1-propenyl > or = 2-methylpropyl > (E)-1-propenyl > or = n-propyl > phenyl > > 2,2-dimethylpropyl. For the 3'-N substituents, the activity decreases in the order t-BuOCO > Ph > n-hexanoyl. A significant increase in the cytotoxicity against the doxorubicin-resistant human breast cancer cell line MCF7-R that expresses the multidrug resistance (MDR) phenotype is observed by the proper modification of the substituent at C-10. The observed remarkable effects of the substituents at C-10 on the activity against MCF7-R can be ascribed to the effective inhibition of the binding of these new taxoids to P-glycoprotein that is responsible for MDR.

Suppressive effects of destruxin B on hepatitis B virus surface antigen gene expression in human hepatoma cells.

Destruxin B, a cyclodepsipeptide was originally identified as a plant pathogen from the fungus, Alternaria brassicae. We examined the antiviral activity of destruxin B and found that it suppresses the expression of the hepatitis B viral surface antigen (HBsAg) gene in human hepatoma Hep3B cells which carry an integrated viral gene in its chromosome. In contrast, destruxin B shows no cytotoxic effect on the viability of the cells. Furthermore, it can be shown that destruxin B can reversibly suppress HBsAg production by Hep3B cells in a concentration-dependent manner with EC50 of 0.5 microM. Northern blot analysis indicates that the suppression of HBsAg gene expression by destruxin B is mainly at the mRNA level. Destruxin B not only suppresses the endogenously expressed HBsAg in the Hep3B cells but also suppresses the HBsAg produced either from the stable transfected HBV DNA in another human hepatoma HuH-7 cell line which carry no endogenous HBV genome. These results suggest that destruxin B may have future potential for development as a specific anti-HBV drug.

A clinical study of schizophrenic suicides. 42 cases in Taiwan.

A clinical study of 25 male and 17 female schizophrenic suicides is presented. Jumping from a height was the most common method used. The 42 suicidal schizophrenics in Taiwan were compared with both 84 sex- and age-matched and 60 5-year illness course non-suicidal schizophrenic control groups. The suicidal schizophrenics were not significantly different from the non-suicidal counterparts of both control groups with regard to age, sex, ethnicity, religion, educational background, the presence of suicide cases in the family history, and the presence of insight, but were significantly different in characteristics of a history of previous suicide attempts, presence of psychotic symptoms during their final month, depression during their final month, a history of depression, a history of previous psychiatric hospitalizations, and the number of hospitalizations. We discuss the findings from this study and others in the literature in the context of the different clinical and socio-cultural backgrounds of these Taiwanese schizophrenic suicides.

Recent advances in liquid-phase combinatorial chemistry.

In combination with high throughput screening, combinatorial organic synthesis of large numbers of pharmaceutically interesting compounds may revolutionize the drug discovery process. Although combinatorial organic synthesis on solid supports is a useful approach, several groups are focusing their research efforts on liquid-phase combinatorial synthesis by the use of soluble polymer supports to generate libraries. This macromolecular carrier, in contrast to an insoluble matrix, is soluble in most organic solvents and has a strong tendency for precipitation in particular solvents. Liquid-phase combinatorial synthesis is a unique approach since homogeneous reaction conditions can be applied, but product purification similar to the solid-phase method can be carried out by simple filtration and washing. This method combines the positive aspects of classical solution-phase chemistry and solid-phase synthesis. This review examines the recent applications (1995-1999) of soluble polymer supports in the synthesis of combinatorial libraries.

Studies on the cytotoxic mechanisms of ginkgetin in a human ovarian adenocarcinoma cell line.

The cytotoxic effects of ginkgetin, a natural biflavone isolated from Selaginella moellendorffii Hieron, were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in three different human cell lines: ovarian adenocarcinoma (OVCAR-3), cervical carcinoma (HeLa) and foreskin fibroblast (FS-5). The concentrations of ginkgetin required to induce 50% death (EC50) in OVCAR-3, HeLa, and FS-5 were 3.0, 5.2, and 8.3 microg/ml, respectively. Morphological changes in cells and their nuclei, DNA fragmentation with a characteristic pattern of inter-nucleosomal ladder, and double-stranded DNA breaks were detected following treatment with 3 microg/ml of this biflavone for 24 h. Incubation with 5 microg/ml ginkgetin led to increased intracellular levels of hydrogen peroxide as early as 30 min. The cytotoxicity of ginkgetin was partially inhibited by pretreating cells with vitamin C, vitamin E or catalase. Catalase not only afforded the best protective effect among three antioxidants, but also reduced both the DNA fragmentation and double-stranded DNA breakage induced by ginkgetin. Moreover, the involvement of caspase(s) in ginkgetin-induced apoptosis was demonstrated by the activation of caspase 3 after drug treatment and the suppression of cell death by a broad-spectrum caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk). However, the protective effects of z-VAD-fmk and catalase were not additive. Taken together, our results indicated that the apoptosis induced by ginkgetin (especially at 5 microg/ml) is mediated mainly through the activation of caspase(s) by the hydrogen peroxide generated possibly through autooxidation of this biflavone.

A tumor cell growth inhibitor from Polygonum hypoleucum Ohwi.

Polygonum hypoleucum Ohwi (P. hypoleucum Ohwi) has been used as a Chinese medicine for a long time. In the present study, four anthraquinones, emodin, emodin 1-O-beta-D-glucoside (49A), physcion (62A), and physcion 1-O-beta-D-glucoside (50A) were identified from P. hypoleucum Ohwi and their inhibitory effects on various tumor cells proliferation were investigated. On a percentage basis, emodin had the highest suppressing activity on the various tumor cells proliferation. At 10 microg/ml, the percentage inhibition on K562 cells proliferation for emodin, 49A, 62A, and 50A were 97+/-3.4%, 18+7.3%, 24+/-3.6%, and 31+/-8.9%, respectively. However, inhibitory activities of 10 microg/ml of emodin, 49A, 62A, or 50A on Raji cells proliferation were 98+/-5.0%, 25+/-5.0%, 22+/-3.2%, and 28+/-4.3%, respectively. It was also found that the both C1 and C3 positions of emodin were important for antitumor action. The IC50s of emodin, 49A, 62A, and 50A on various tumor cells were also calculated. The IC50 of emodin on K562 cells was significantly lower than on Raji, HeLa, Calu-1, Wish, and Vero cells (1.5+/-0.2 vs. 2.8+/-0.4 microg/ml, P < 0.01 ;1.5+/-0.2 vs. 8.4+/-1.6 microg/ml; 1.5+/-0.2 vs. 8.9+/-1.0 microg/ml; 1.5+/-0.2 vs. 8.7+/-0.5 microg/ml; 1.5/-0.2 vs. 3.5+/-0.12 microg/ml; P < 0.001). The results indicated that K562 and Raji cells were more sensitive to emodin treatment. Cell viability test indicated that inhibitory effect of emodin on various tumor cell lines was not through direct cytotoxicity. It suggested P. hypoleucum Ohwi included a tumor cell growth inhibitor.

Blocking of cell proliferation, cytokines production and genes expression following administration of Chinese herbs in the human mesangial cells.

In the hope of identifying agents of therapeutic value in immuoglobulin A nephropathy (IgA-N), we tested crude methanol extracts of 15 Chinese herbs for their effect on human mesangial cell proliferation. The results indicated that 4 out of the 15 crude extracts inhibited human cells proliferation activated by IL-1beta and IL-6. The extracts and their median inhibitory concentrations were as follows (in microg/ml): Ludwiga octovalvis (MLS-052), 49.9 +/- 1.8; Rhus semialata (MLS-053), 31.2 +/- 1.6; Tabernaemontana divaricata (MLS-054), 50.0 +/- 2.1; Amepelopsis brevipedunculata (MLS-059), 42.9 +/- 1.1. These findings indicate that human mesangial cells were most sensitive to MLS-053 treatment. These herbs also decreased interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) production. Moreover, IL- 1beta mRNA expression was inhibited by Rhus semialata (R. semialata; MLS-053). It is unlikely that cytotoxicity was involved, because no cell deaths were observable. We hypothesize that the inhibitory mechanisms of these Chinese herbs may be related to the impairments of gene expression and production of cytokines in human mesangial cells. Plans are underway for the isolation of pure compounds from these Chinese herbs and the elucidation of their mechanisms of action.

Portal hypotensive effects of tetrandrine and verapamil in portal hypertensive rats.

The portal hypotensive effects of tetrandrine and verapamil (both calcium-channel blockers) were assessed in portal hypertensive rats. Portal hypertension was induced by partial portal vein ligation in Sprague-Dawley rats. Both tetrandrine (4, 8, 16 and 24 mg kg-1) and verapamil (0.5, 1.0, 1.5 and 2.0 mg kg-1) induced dose-dependent decreases of portal venous pressure and mean arterial pressure after intravenous infusion. for example, infusion of tetrandrine (16 mg kg-1) induced a maximum reduction of portal venous pressure and mean arterial pressure approximately 1 min after the start of infusion. Portal venous pressure decreased from baseline (12.5 mmHg) to 10.0 mmHg, and mean arterial pressure from baseline (90 mmHg) to 80 mmHg. Heart rate decreased from 250 to 240 beats min-1. At 24 mg kg-1, tetrandrine reduced portal venous pressure and mean arterial pressure to 20.3 +/- 2.4% and 28.4 +/- 1.4% of baseline, respectively. Our results show that both tetrandrine and verapamil induce portal pressure reduction in portal hypertensive animals.

[Clinical study of rapid bladder filling agent].

The Rapid Bladder Filling Agent (RBFA) was prepared with Polyporus umbellatus, Poria cocos and Furosemidum. The urinary volume and the best filling time of urinary bladder were observed with ultrasonography in 211 cases. The result showed that in experimental group the largest urinary volume in unit time was more than that in control group and shortest filling time was shorter (30.35 +/- 7.9 min) in comparing with control group (145.6 +/- 50.1 min). Clinical observation for 1180 cases proved that the RBFA had the effects of raising the quality of ultrasonographic examination and escalating work efficiency, shortening the waiting time of patients and relieving patients' suffering from excessive filling of urinary bladder. The effective time of the RBFA was fast but the duration was short. The RBFA had no adverse effect.

Tumor necrosis factor alpha increases epithelial barrier permeability by disrupting tight junctions in Caco-2 cells.

The objectives of this study were to determine the effect of tumor necrosis factor alpha (TNF-alpha) on intestinal epithelial cell permeability and the expression of tight junction proteins. Caco-2 cells were plated onto Transwell microporous filters and treated with TNF-alpha (10 or 100 ng/mL) for 0, 4, 8, 16, or 24 h. The transepithelial electrical resistance and the mucosal-to-serosal flux rates of the established paracellular marker Lucifer yellow were measured in filter-grown monolayers of Caco-2 intestinal cells. The localization and expression of the tight junction protein occludin were detected by immunofluorescence and Western blot analysis, respectively. SYBR-Green-based real-time PCR was used to measure the expression of occludin mRNA. TNF-alpha treatment produced concentration- and time-dependent decreases in Caco-2 transepithelial resistance and increases in transepithelial permeability to the paracellular marker Lucifer yellow. Western blot results indicated that TNF-alpha decreased the expression of phosphorylated occludin in detergent-insoluble fractions but did not affect the expression of non-phosphorylated occludin protein. Real-time RT-PCR data showed that TNF-alpha did not affect the expression of occludin mRNA. Taken together, our data demonstrate that TNF-alpha increases Caco-2 monolayer permeability, decreases occludin protein expression and disturbs intercellular junctions.