Treatment of an incarcerated inguinal hernia associated with abdominal tuberculosis in an adult patient.

The incidence of tuberculosis is increasing worldwide, especially in developing countries. The prevalence of abdominal tuberculosis has been found to be as high as 12% in people with extrapulmonary tuberculosis. Peritoneal thickening and intestinal adhesions can occur in patients with abdominal tuberculosis. Inguinal hernias are extremely rare in people with abdominal tuberculosis; only 11 cases have been reported in the English-language literature, half of which involved pediatric patients. No definitive guideline on the management of such cases is available. In this report, we describe the unusual finding of an incarcerated inguinal hernia in an adult with abdominal tuberculosis and propose a therapy to treat this complicated disease based on our successful experience.

[Effect of glucose analogs on bofilm and expressions of related genes in Staphylococcus epidermidis].

Staphylococcus epidermidis, an opportunistic human pathogen, has become the most important cause of nosocomial infections in recent years. Its infection is mainly due to the ability to form biofilm on indwelling medical devices. To investigate the response mechanism of S. epidermidis to environment in biofilm formation and find efficient anti-biofilm methods, we investigated effects of two glucose analogs, 2-Deoxy-D-glucose (2DG) and Methyl-D-glucoside (MG), on biofilm formation, expression of related gene and changes of surface protein in S. epidermidis 97-337 with high biofilm formation capability and pathogenicity. The effect of MG on biofilm formation was more complex than that of 2-DG which is a strong inhibitor in S. epidermidis 97-337 growth. MG can induce biofilm formation of S. epidermidi 97-337 in low concentration and exhibited strong inhibition only in high concentration, and distinctly inhibited the primary attachment to poly-material. In S. epidermidi 97-337 cultured in media with MG, expressions of ica and AtlE were not be changed obviously in mRNA level, but mRNA expression of agr gene increased distinctly, and MG disturbed component of surface proteins of S. epidermidi 97-337. Glucose analogies MG can inhibit S. epidermidi 97-337 biofilm formation, and MG inhibition in initiating attachment dramatically contributes to it. MG inhibition effects result not from regulating the expression of ica and AtlE genes but from changing the protein components on surface by regulating agr gene expression, and it can be presumed that MG's competitive character in bacteria glycose metabolism is crucial factor for these effects.

[Effect of glucose on biofilm and the gene ica expression in Staphylococcus epidermidis with different biofilm-forming capability].

The infection of S. epidermidis, an opportunistic human pathogen, depends on biofilm formation, and biofilm formation is closely related to environment. Researches in the thesis focused on two strains of S. epidermidis with different capability of biofilm formation. To find the mechanism of response to environment on biofilm formation, biofilm formation and expression of ica, icaR, AtlE in theses S. epidermidis cultivated in different grow environment and in media with glucose for different time were assayed. Glucose can induce the biofilm formation by inducing ica gene, but the inducing do not need continued ica expression, and other genes also contribute to the regulation; anti-ODN specially binding icaADBC can withstand biofilm inducing from glucose. These results suggest that biofilm formation closely related to growth environment, which is a complex regulation mechanism. Biofilm formation is closely related to bacteria energy metabolism and cell wall synthesis. Some crucial factors in the complex and integrated regulation system have not been known yet.

Integrin beta1 subunit overexpressed in the SMMC-7721 cells regulates the promoter activity of p21(CIP1) and enhances its transcription.

Evidence has been emerging to suggest that integrin could induce growth inhibition in some cell types. Some of the molecular mechanisms underlying growth arrest have been elucidated. We reported here that overexpression of integrin beta1 imposed a growth inhibitory effect on the hepatocellular carcinoma cell line SMMC-7721, and this phenomenon was mainly attributed to the cyclin-dependent kinase inhibitor p21(CIP1). Furthermore, our findings suggested that transcription activity of the p21(CIP1) gene could be upregulated in the integrin beta1-overexpressing cells, and possibly controlled by the cis-elements in the core region of the p21(CIP1) promoter.

Role of FAK in TNF-alpha/Cycloheximide-induced Apoptosis of SMMC-7721 Cells.

To explore the role of FAK in TNF-alpha/cycloheximide-induced apoptos is of human hepatocellular carcinoma cell line SMMC-7721, the FAK antisense plasmid was constructed and transfected into SMMC-7721 cells. Western blot assay was adopted to examine PKB level. Flow cytometry assay was used to detect apoptosis. It was shown that the SMMC-7721 cells were insensitive to TNF-alpha cytotoxicity, but they entered apoptosis quickly in the presence of cycloheximide and TNF-alpha. PKB was decreased during TNF-alpha/cycloheximide-induced apoptosis. No significant change of PKB level was found in the presence of TNF-alpha or cycloheximide, respectively, seeming that PKB level was closely correlated with apoptosis. When FAK was 60% reduced as a result of the transfection of SMMC-7721 cells with FAK antisense construct, the percentage of TNF-alpha/cycloheximide-induced apoptosis was enhanced at lower dose of TNF-alpha but decreased at higher dose of TNF-alpha, compared with the control. Correspondingly, the PKB level in FAK-down-regulated transfectants was lower at lower dose of TNF-alpha, but higher at higher dose of it. Therefore, FAK regulated TNF-alpha/cycloheximide-induced apoptosis in a biphase manner. This function might be related with PKB level.

[The effects of PTEN gene on migration and FAK phosphorylation of SMMC-7721 human hepatocarcinoma cell line].

PTEN is a major tumor suppressor gene that encodes a dual-specificity phosphatase with high sequence similarity to the cytoskeletal protein tensin. PTEN may be involved in the formation and disassembly of focal adhesion and affect cell migration. In the present study, PTEN expression plasmid was constructed and transfected into the hepatoma cell line SMMC-7721 to analyze the alterations of cell motility and FAK tyrosine phosphorylation. It was observed that the overexpression of PTEN gene significantly inhibited cell motility on extracellular matrix (Fn), and the cell migration on fibronectin was reduced by 35%. Similarly, at 30-min and 60-min, the cell spreading on Fn but not on polylysine was inhibited by 29% and 26% respectively. The data obtained from immunoprecipitation and immunoblotting analyses showed that the overexpression of PTEN did not affect FAK expression but resulted in a decrease in FAK tyrosine phosphorylation. The level of FAK phosphorylation was inversely correlated with the level of PTEN protein in three cell lines. It was also found that the overexpression of PTEN led to growth inhibition, with the number of cells in S phase reduced by 16%. These results indicate that PTEN exerts its tumor-suppressive effects on hepatocellular carcinoma cells through the inhibition of cell motility and cell cycle progression.

Targeted deletion of the mitogen-activated protein kinase kinase 4 gene in the nervous system causes severe brain developmental defects and premature death.

The c-Jun NH2-terminal protein kinase (JNK) is a mitogen-activated protein kinase (MAPK) involved in the regulation of various physiological processes. Its activity is increased upon phosphorylation by the MAPK kinases MKK4 and MKK7. The early embryonic death of mice lacking an mkk4 or mkk7 gene has provided genetic evidence that MKK4 and MKK7 have nonredundant functions in vivo. To elucidate the physiological role of MKK4, we generated a novel mouse model in which the mkk4 gene could be specifically deleted in the brain. At birth, the mutant mice were indistinguishable from their control littermates, but they stopped growing a few days later and died prematurely, displaying severe neurological defects. Decreased JNK activity in the absence of MKK4 correlated with impaired phosphorylation of a subset of physiologically relevant JNK substrates and with altered gene expression. These defects resulted in the misalignment of the Purkinje cells in the cerebellum and delayed radial migration in the cerebral cortex. Together, our data demonstrate for the first time that MKK4 is an essential activator of JNK required for the normal development of the brain.

Prohibitin suppresses renal interstitial fibroblasts proliferation and phenotypic change induced by transforming growth factor-beta1.

Prohibitin (PHB), a potential tumor suppressor, has been shown to inhibit cell proliferation by repressing E2F-mediated transcription. But little is known about the role of PHB involved in tubulointerstitial fibrosis (TIF). Here, for the first time, we found PHB protein was positively expressed at normal renal tissues, strongly down-regulated in renal biopsy specimens, and negatively correlated with the expression of alpha-smooth-muscle actin (alpha-SMA) and with the degrees of tubulointerstitial lesions. Transforming growth factor-beta1 (TGF-beta1) is the most important profibrotic cytokine in the process of TIF and capable of inducing cell phenotypic change of interstitial fibroblasts characterized by the de novo expression of alpha-SMA. Confocal microscopy showed majority of PHB is located at cytoplasm as well as at nucleus in rat kidney fibroblasts cell (NRK-49F). As we found that PHB protein and mRNA expression were down-regulated in NRK-49F cells following TGF-beta1 stimulation. We used transient transfection to over-express PHB protein and found that cells with increased PHB levels had a significant reduction in the percentage entering cell cycle and abolished de novo expression of alpha-SMA following TGF-beta1 stimulation. Therefore, over-expression of PHB suppresses renal interstitial fibroblasts proliferation and cell phenotypic change induced by TGF-beta1, which indicates PHB as a potential therapeutic target to halt the progression of TIF.

Integrin beta(1A) upregulates p27 protein amount at the post-translational level in human hepatocellular carcinoma cell line SMMC-7721.

Integrins mediate many fundamental cellular processes by binding to components of the extracellular matrix. We showed previously that integrin beta(1A) could inhibit cell proliferation. Integrin beta(1A) stimulated the promoter activity of p21(cip1) and enhanced its transcription in SMMC-7721 cells. In this study, we demonstrated that integrin beta(1A) upregulated p27(kip1) at the post-translational level in SMMC-7721 cells. Our results showed that integrin beta(1A) increased the p27 protein amount, both in cytoplasm and nucleus, but did not affect the p27 mRNA amount. Cycloheximide treatment experiment revealed that the half-life of p27 protein was prolonged in integrin beta1A overexpressing cells, indicating that integrin beta(1A) inhibited the degradation of p27 protein. Our data also provided evidence that both the proteasome and calpain were involved in the degradation of p27 protein in SMMC-7721 cells. Integrin beta(1A) decreased the Skp2 expression and repressed the activity of calpain during G1 phase in SMMC-7721 cells. Taken together, these results indicated that integrin beta(1A) might upregulate the protein amount of p27 through repressing Skp2-dependent proteasome degradation and calpain-mediated proteolysis in SMMC-7721 cells.

Profilin 1 obtained by proteomic analysis in all-trans retinoic acid-treated hepatocarcinoma cell lines is involved in inhibition of cell proliferation and migration.

Previous studies have shown that all-trans retinoic acid (ATRA) suppresses growth of hepatocarcinoma cell in vitro. To understand the underlying mechanisms, we investigated the protein expression profiles by 2-DE in hepatocarcinoma cell line SMMC-7721 treated with ATRA. Our results reveal that six proteins were differently expressed in response to ATRA. Using MS and database searching, they were identified as profilin 1, phosphoglycerate kinase 1, RuvB-like 1, alpha-enolase, pyridoxal kinase and F-actin capping protein. We selected the up-regulated protein, profilin 1 (PFN1), for further studies. The PFN1 expression was increased in response to ATRA in a dose- and time-dependent manner. The PFN1 expression was reduced dramatically in four hepatoma cell lines compared to L02 cell line of non-tumor origin. The PFN1 expression was also examined in 4 cases of primary hepatocarcinoma tissues by Western blot and 30 cases by tissues microarray. It was found that the protein level of PFN1 was lower in hepatocarcinoma tissues compared to that in the adjacent tissues. Similar to ATRA, overexpression of PFN1 led to inhibition of cell proliferation and migration. Furthermore, RNAi-based PFN1 knockdown could rescue the inhibitory effect of ATRA on cell proliferation and migration. In conclusion, ATRA inhibited cell proliferation and migration through up-regulation of PFN1.

Protein phosphatase activity of PTEN inhibited the invasion of glioma cells with epidermal growth factor receptor mutation type III expression.

PTEN is a major tumor suppressor gene that has been shown to inhibit cell invasion. Its mutation has been found in 20-40% of malignant gliomas. Meanwhile, the type III EGFR mutation (EGFRvIII), which was frequently found in gliomas, promoted cell invasion. In the present study, the effects of PTEN on cell invasion were investigated in U87DeltaEGFR glioblastoma cells with EGFRvIII expression but missing PTEN. The cell invasion was downregulated by transfection of phosphatase-active forms of PTEN (wild-type and G129E) but not by PTEN (C124A) with an inactive phosphatase domain; the effects were correlated with decreased tyrosine phosphatase levels of FAK at Tyr397, which was increased by EGFRvIII. Overexpression of FAK mutant (Y397F) could partially mimic the effect of PTEN on cell invasion. Although EGFRvIII increased the levels of P-Akt and PTEN eliminated it, PI-3K inhibitors, wortmannin or Ly294002, could not decrease the cell invasion. In conclusion, PTEN could inhibit cell invasion even in the presence of the constitutively active EGFR; this inhibition depended on its protein phosphatase activity, partially by dephosphorylating FAK, but not depended on its lipid phosphatase activity.