Downregulation of AlphaB-crystallin in Retinal Pigment Epithelial Cells Exposed to Diabetes-related Stimuli In Vivo and In Vitro.

BACKGROUND/AIM: AlphaB-crystallin plays a pivotal role in many diseases. However, the involvement of alphaB-crystallin in retinal pigment epithelial (RPE) cells with diabetes stimuli remains unknown. The aim of this study is to examine the alterations of RPE cells and alphaB-crystallin expression in diabetic models in vivo and in vitro. MATERIALS AND METHODS: Diabetic conditions in mice were induced by streptozotocin (STZ). The thickness of the RPE/choroid complex was measured by optical coherence tomography (OCT). Periodic acid-Schiff (PAS) staining was used to investigate the choriocapillaris in histological sections of murine eyeballs and oxidative stress was evaluated using immunofluorescence with anti-4-hydroxynonenal (HNE) antibody. AlphaB-crystallin expression was examined in the RPE/choroid complex using ELISA. Real-Time PCR was performed to evaluate the alphaB-crystallin expression in cultured human RPE cells with high glucose or following advanced glycation end-products (AGE) stimulation. RESULTS: In diabetic mice, OCT-based RPE/choroidal layers were thickened 2 months after STZ stimulation, where PAS-positive dilated choriocapillaris was noted. Immunoreactivity of 4-HNE was strongly observed in the RPE layer, from which a significant number of RPE cells was lost. Meanwhile, alphaB-crystallin expression in 2-month STZ mice was significantly lower compared to controls. In accordance with these results, in vitro data showed that the alphaB-crystallin expression was also significantly lower in RPE cells with high glucose or following AGE stimulation compared to untreated cells. CONCLUSION: In both types of diabetic models the expression of alphaB-crystallin was found to be downregulated in RPE cells and was associated with increased levels of oxidative stress.

CD28 deficiency attenuates primary blast-induced renal injury in mice via the PI3K/Akt signalling pathway.

INTRODUCTION: Primary blast affects the kidneys due to direct shock wave damage and the production of proinflammatory cytokines without effective treatment. CD28 has been reported to be involved in regulating T cell activation and secretion of inflammatory cytokines. The aim of this study was to investigate the influence of primary blast on the kidney and the effect of CD28 in mice. METHODS: A mouse model of primary blast-induced kidney injury was established using a custom-made explosive device. The severity of kidney injury was investigated by H&E staining. ELISA was applied to study serum inflammation factors' expression. Western blot assays were used to analyse the primary blast-induced inflammatory factors' expression in the kidney. Immunofluorescence analysis was used to examine the PI3K/Akt signalling pathway. RESULTS: Histological examination demonstrated that compared with the primary blast group, CD28 deficiency caused a significant decrease in the severity of the primary blast-induced renal injury. Moreover, ELISA and western blotting revealed that CD28 deficiency significantly reduced the levels of interleukin (IL)-1ß, IL-4 and IL-6, and increased the IL-10 level (p<0.05). Finally, immunofluorescence analysis indicated that PI3K/Akt expression also changed. CONCLUSIONS: CD28 deficiency had protective effects on primary blast-induced kidney injury via the PI3K/Akt signalling pathway. These findings improve the knowledge on primary blast injury and provide theoretical basis for primary blast injury treatment.

Analysis on difference in gastrointestinal hormone levels of patients with the history of diabetes and concurrent nephropathy and study on the role of liraglutide.

OBJECTIVE: To compare the difference in the gastrointestinal hormone levels of the patients with the history of diabetes and concurrent nephropathy and investigate the clinical effect of liraglutide in the treatment of diabetic nephropathy (DN). PATIENTS AND METHODS: 42 cases of patients with DN admitted in our hospital from April 2010-May 2015 were selected and divided into phase I-II group (group A, n = 22) and phase III-IV group (group B, n = 20) according to DN phases and 20 cases of patients with diabetes rather than nephropathy admitted in our hospital during the same period were selected as the control group, all of whom underwent the routine biochemical test and gastrointestinal hormone test, the differences in gastrin (GAS), motilin (MTL) and glucagon (GLC) of DN patients were compared at different phases, the gastric emptying test was carried out on them and the gastric emptying time was recorded. All patients were treated with liraglutide and the changes in fasting blood glucose (FBG), glycosylated hemoglobin (HbAlc), serum creatinine (Cr), blood urea nitrogen (BUN), insulin (FINS) and insulin resistance level (HOMA-IR) were tested before treatment and after 10 weeks' treatment, the changes in the tumor necrosis factor (TNF-α), interleukin -6 (IL-6) and transforming growth factor (TGF-ß1) were determined, and the change in the gastrointestinal hormone levels of patients was recorded after treatment. RESULTS: (1) the GAS, MTL, GLC and gastric emptying time in group B were higher than those in group A and the control group (p < 0.05), and the above indicators in group A were higher (p < 0.05); (2) after 10 weeks' treatment, the gastrointestinal hormone levels in the three groups were reduced and the gastric emptying time was shortened, the difference was statistically significant (p < 0.05) compared with those before treatment, after 10 weeks' treatment, the GAS, MTL, GLC and gastric emptying time in group B were higher than group A and the control group, those in group A were higher than control group (p < 0.05); (3) before treatment, the comparative differences in FBG, HbAlc, FINS and HOMA-IR among the three groups were not statistically significant (p > 0.05), and after 10 weeks' treatment, the differences in FBG, HbAlc and HOMA-IR among three groups were reduced and FINS was increased, the difference in those between before treatment and after treatment was statistically significant (p < 0.05) and the comparative difference among the three groups was not statistically significant (p > 0.05); (4) before treatment, Cr and BUN levels in group A and group B were higher than the control group (p < 0.05), after 10 weeks' treatment, the Cr and BUN levels among three groups were significantly decreased (p < 0.05), Cr and BUN in group A and group B were higher than the control group, cr and BUN levels in group B were higher than group A (p < 0.05); (5) before treatment, the difference by comparing IL-6, TNF-α and TGF-ß1 among three groups were not statistically significant (p > 0.05), after 10 weeks' treatment, the indicators in the three groups were decreased significantly (p < 0.05), but the comparative difference among the three groups were not statistically significant (p > 0.05); (6) the difference by comparing the efficiencies among the three treatment was not statistically significant (p > 0.05). CONCLUSIONS: There are some correlations between the gastrointestinal hormone levels and the degree of renal impairment of DN patients. Good results will be achieved by applying liraglutide in intervention with different phases of DN and DM patients, which cannot only regulate the gastrointestinal hormone levels and lower the blood sugar levels of patients, but can also reduce the insulin resistance and delay the process of renal damage.

Association between expression of DNA mismatch repair genes and clinical features and prognosis of patients with radical resection of colon cancer.

The aim of this study was to investigate the clinical significance of the expression of DNA mismatch repair (MMR) genes in patients subjected to radical surgical removal of colon cancer, as well as their correlation with disease prognosis. Ninety stage II and III colon cancer patients who received laparoscopic radical resection of colon cancer at our hospital were recruited in this study. The expression of hMLH1, hMSH2, hMSH6, and hPMS2 in the resected tumor tissues was examined by SP immunohistochemistry, in order to analyze the relationship between defective DNA MMR (dMMR) and the clinico-pathological features and prognosis of colon cancer. Patients were followed up over a period of 5-35 months, and the Kaplan-Meier survival curve was plotted. dMMR was confirmed in 27 subjects (30.0%), among whom recurrence with metastasis and death was reported in 5 (18.5%) and 2 (7.4%) patients, respectively. The remaining 63 subjects displayed proficient DNA MMR (pMMR); among these, 19 (30.2%) and 7 (11.1%) recurrences with metastasis and death were reported, respectively. dMMR showed no significant correlation with gender, age, or therapeutic modality (P > 0.05), but was significantly correlated with the degree of differentiation, tumor location, number of resected lymph nodes, presence of ileus, and TNM stage (P < 0.05). The prognosis of patients with dMMR was better than that of patients with pMMR. dMMR serves as a biomarker for the prognosis of stage II/III colon cancers.

Influence of melatonin on IL-1Ra gene and IL-1 expression in rats with liver ischemia reperfusion injury.

The aim of the present study was to explore the influence of melatonin (MT) on rats with liver ischemia reperfusion injury (IRI) and its mechanism. A total of 66 male Sprague-Dawley rats were randomly divided into 3 groups: i) Normal control group, ii) ischemia reperfusion group (IR group) and iii) melatonin treatment group (MT group). Rats in the MT group were administered an intraperitoneal injection of MT (10 mg/kg, 1 ml) at 70 and 35 min before ischemia, early reperfusion, and 1 and 2 h after reperfusion, respectively. Blood was removed at the normal time-point (prior to any processes), 35 min before ischemia, 2, 4, 8 and 24 h after reperfusion. Subsequently the rats were sacrificed. The pathological changes of liver tissues, interleukin-1 receptor antagonist (IL-1Ra) gene and IL-1 expression levels were detected. There were no evident differences between the immediate reperfusion and 2 h IR group and MT group. The liver structure injury of the 4, 8 and 24 h MT groups were improved to various differences compared to the corresponding IR group; liver IL-1ß of the MT group at 35 min after ischemia, and 2, 4, 8 and 24 h after reperfusion was evidently lower than that of the IR group (P<0.05); IL-1Ra mRNA expression in the 2 h MT group was higher compared to the 2 h IR group by 4.85-fold; and IL-1Ra mRNA expression in the 4 h MT group was higher compared to the 4 h IR group by 9.34-fold. Differences between two groups at other time-points were <2-fold. In conclusion, MT can upregulate IL-1Ra gene expression by reducing generation of IL-1 thus reducing IRI.

Effects of melatonin on liver function and lipid peroxidation in a rat model of hepatic ischemia/reperfusion injury.

The present study aimed to investigate the effects of melatonin (MT) on liver function and lipid peroxidation following hepatic ischemia-reperfusion injury (IRI). A total of 66 male Sprague-Dawley rats were randomly assigned into three groups: Normal control (N) group, ischemia-reperfusion (IR) group and the MT-treated group. A hepatic IRI model was developed by blocking the first porta hepatis, and subsequently restoring hepatic blood inflow after 35 min. Following reperfusion, changes in the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were detected by a chemical method at various time points. In the MT group, the MDA levels were significantly reduced (P<0.05) at all time points, as compared with the IR group. Furthermore, SOD activity was significantly increased (P<0.05) in the MT group, as compared with the IR group at all time points; and the levels of GSH in the MT group were significantly higher (P<0.05) than those of the IR group at 2, 4, and 8 h post-reperfusion. The levels of ALT, AST and LDH were significantly reduced in the MT group at each time point, as compared with that of the IR group (P<0.05). In conclusion, MT exhibits potent antioxidant properties that may create favorable conditions for the recovery of liver function following IRI.

Influence of continuous intervention on growth and metastasis of human cervical cancer cells and expression of RNAmiR-574-5p.

This study was carried out to acquire solid evidence that some common treatments could affect micro ribonucleic acids (miRNAs) by revealing the regulatory effect of genes, so as to provide a reference for further exploration of the prevention and treatment of cervical cancer. Nude mouse tumorigenicity assay was used to study the effect of inhibiting miR-574-5p on development and tumorigenic ability of Henrietta Lacks (HeLa) tumor. Cell wound scratch assay, flow cytometry and real-time quantitative polymerase chain reaction (RT-qPCR) were adopted to study the effects of anoxia and temperature, etc., on expression of miR-574-5p and QKI in HeLa as well as on the clone and migration ability of cells, to provide prevention and treatment of cervical cancer with new ideas and evidence. The results demonstrated that cervical cancer tissues had a significantly increased miR-574-5p expression compared with para-carcinoma tissues; conversely, Gomafu, overall QKI (pan-QKI) and QKI-5 messenger ribonucleic acid (mRNA) and protein expression all decreased. Part of the common nursing methods had a certain influence on miR-574-5p expression, HeLa reproduction and metastasis, and even cell cycle. For example, ultraviolet (UV) irradiation was effective in decreasing miR-574-5p expression of HeLa and inhibiting cell migration; severe hypoxia significantly decreased the survival rate of HeLa, leading to the increase of programmed death percentage and cell ratio in G2/M phase as well as the decrease of cell ratio in G1 phase. Incubation at different temperatures also affected miR-574-5p expression and cell proliferation. Thus, it can be known that miR-574-5p, Gomafu and QKI expression in cervical cancer tissues and para-carcinoma tissues are significantly up-regulated or down-regulated. Some treatments, such as UV irradiation, hypoxia, incubation temperatures, etc., can affect miR-574-5p expression and HeLa proliferation as well as metastases in different degrees. These findings provide a reference and basis for further study.

The epidemiology and surgical outcomes of patients undergoing primary total hip replacement: an Asian perspective.

INTRODUCTION: The number of total knee replacements far exceeds the number of primary total hip replacements (THR) in Singapore. There is a paucity of data regarding patients who undergo THR in Singapore. This paper aimed to study the epidemiology and early postoperative outcomes of patients receiving primary THR in a single tertiary institution. METHODS: This is a retrospective study of all patients undergoing primary THR between January 2003 and December 2005. The following variables were analysed: patient demographics, surgical approach, mode of implant fixation, postoperative complications and functional outcomes using D'Aubigne and Postel scoring. RESULTS: There were a total of 115 patients who underwent primary THR over a 36-month period. The mean age of patients was 55 (range 23-80) years. The male-to-female ratio was 1:2. The most common diagnoses in descending order were as follows: inflammatory arthritis, osteoarthritis, avascular necrosis, hip dysplasia and post-traumatic osteoarthritis. Patients were evaluated at a mean follow-up of 41 months, with 90.8 percent having excellent and good outcomes. There were no statistical differences between the surgical approach and implant fixation with regard to postoperative outcome. CONCLUSION: In Asian patients, the three commonest aetiologies for degenerative hip arthritis were inflammatory arthropathies, primary osteoarthritis and avascular necrosis. Regardless of diagnosis, the patient groups had comparable functional outcomes following THR.

Alpha-4/beta-1 and alpha-L/beta-2 integrins mediate cytokine induced lung leukocyte-epithelial adhesion and injury.

BACKGROUND AND PURPOSE: Injury to the alveolar epithelium is a critical feature of acute lung injury (ALI). Using a cytokine model of ALI we demonstrated previously that newly recruited mononuclear phagocytes (MNP) contributed to lung inflammation and injury. We hypothesized that cytokines delivered into the alveolar airspace would have multiple effects on the lung that may contribute to lung injury. EXPERIMENTAL APPROACH: Intratracheal cytokine insufflation and leukocyte adoptive transfer in vivo were combined with in vitro analyses of lung epithelial cell-MNP adhesion and injury. Lung inflammatory injury was assessed by histology, leukocyte infiltration, and release of LDH and RAGE. KEY RESULTS: Cytokine insufflation was associated with apparent MNP-epithelial adhesion, up-regulation of alveolar ICAM-1 and VCAM-1, and the release of LDH and RAGE into the bronchoalveolar lavage. Insufflation of small molecule integrin antagonists suppressed adhesion of MNP and modulated release of LDH and RAGE. Adoptive transfer of MNP purified from cytokine insufflated lungs into leukopenic rats demonstrated the requirement of MNP for release of LDH that was not induced by cytokine alone. Corroboration that disrupting the ICAM/LFA1 interaction or the VCAM/VLA4 interaction blocked MNP-epithelial cell interaction and injury was obtained in vitro using both blocking monoclonal antibodies and the small molecule integrin antagonists, BIO5192 and XVA143. CONCLUSIONS AND IMPLICATIONS: MNP recruited following cytokine insufflation contributed to lung injury. Further, integrin antagonists reduced alveolar epithelial cell injury induced during lung inflammation. Intratracheal delivery of small molecule antagonsists of leukocyte-epithelial adhesion that prevent lung injury may have significant clinical utility.

Chaperoning of estrogen receptor and induction of mammary gland proliferation by neuronal protein synuclein gamma.

Synucleins are emerging as central players in the formation of pathologically insoluble deposits characteristic of neurodegenerative diseases. However, synuclein gamma (SNCG), previously identified as a breast cancer specific gene (BCSG1), is also highly associated with breast cancer progression. Using transgenic mouse model, we demonstrated a role of SNCG in induction of highly proliferative pregnancy-like phenotype of mammary epithelial cells and branching morphology. SNCG participated in the heat shock protein-based multiprotein chaperone complex for steroid receptor signaling. Expression of SNCG in mammary epithelium resulted in a significant stimulation of ERalpha transcriptional activity. SNCG-induced mammary gland proliferation can be effectively blocked by antiestrogen and ovariectomy, indicating that the induced proliferation is mediated by ERalpha signaling and requires estrogen stimulation. These data indicate the chaperone activity of SNCG on stimulation of steroid receptor signaling in mammary gland and, thus induces extensive mammary gland proliferation and contributes to the hormonal impact on mammary tumorigenesis.

Inhibition of Wilms' tumor growth by intramuscular administration of tissue inhibitor of metalloproteinases-4 plasmid DNA.

Extracellular matrix (ECM) degrading matrix metalloproteinases (MMPs) lead to ECM turnover, a key event in cancer growth and progression. The tissue inhibitors of matrix metalloproteinases (TIMPs) limit the activity of MMPs, which suggests their use for cancer gene therapy. Here we report that systemic administration of naked TIMP-4 DNA significantly inhibited Wilms' tumor growth in nude mice. TIMP-4, whose expression was lost in Wilms' tumor, inhibited the growth of G401 Wilms' tumor cells at a concentration lower than those required for MMP inhibition. This inhibition was associated with internalization of exogenous recombinant TIMP-4. Electroporation-mediated intramuscular injection of TIMP-4 expression plasmid resulted in sustained plasma TIMP-4 levels and significant tumor suppression. Our data demonstrate a tumor suppressive effect of TIMP-4 against Wilms' tumor and the potential utility of intramuscular delivery of TIMP gene for treatment of kidney derived cancers.

Social anxiety in Chinese adults with oral-facial clefts.

OBJECTIVE: This study examined social anxiety and measures of psychosocial adjustment in Chinese adults with oral-facial clefts, their unaffected siblings, and age-matched controls. DESIGN: This cross-sectional study utilized a matched case-control study design. PARTICIPANTS: Eighty-five adult cleft lip and cleft palate (CL/CP) subjects and 85 unaffected siblings (one adult sibling of each CL/CP subject) were recruited in Shanghai, China, from a larger CL/CP study. Eighty-five unaffected controls, gender- and age-matched to the CL/CP subjects, were recruited from Shanghai work units including factories, universities, and other institutions. OUTCOME MEASURES: Social Avoidance and Distress Scale, Fear of Negative Evaluation, Rosenberg Self-Esteem Scale, Interpersonal Support Evaluation List. RESULTS: Affected adults reported significantly more social anxiety than unaffected siblings and controls. Affected adults also scored significantly lower on measures of self-esteem and social support than unaffected siblings and controls. Unaffected siblings and controls were not found to differ on any of these measures. CONCLUSIONS: Findings suggest that individuals with oral-facial clefts may be more disadvantaged with respect to social affiliation and adaptation than unaffected adults. Cross-cultural research is essential in enabling us to determine whether similar trends exist across cultures.

Stimulation of mammary tumorigenesis by systemic tissue inhibitor of matrix metalloproteinase 4 gene delivery.

Tissue inhibitors of matrix metalloproteinase (TIMPs) are multifunctional proteins with both matrix metalloproteinase (MMP) inhibitory effects and growth-regulatory activity. TIMPs inhibit MMP activity, suggesting a use for cancer gene therapy. However, here we report that systemic administration of human TIMP-4 by electroporation-mediated i.m. injection of naked TIMP-4 DNA stimulates tumorigenesis of human breast cancer cells in nude mice. Consistent with tumor stimulation, TIMP-4 up-regulates Bcl-2 and Bcl-X(L) protein. TIMP-4 also inhibits apoptosis in human breast cancer cells in vitro and mammary tumors in vivo. A synthetic MMP inhibitor BB-94 did not have such antiapoptotic effect. Analysis of TIMP-4 expression in human mammary specimens indicates that TIMP-4 protein is increased in mammary carcinoma cells compared with normal mammary epithelial cells. These data indicate an antiapoptotic activity in breast cancer cells and a tumor-stimulating effect of TIMP-4 when administrated systemically.

Induction of mammary differentiation by mammary-derived growth inhibitor-related gene that interacts with an omega-3 fatty acid on growth inhibition of breast cancer cells.

We previously identified and characterized a novel tumor growth inhibitor and a fatty acid-binding protein in human mammary gland and named it the mammary-derived growth inhibitor-related gene (MRG). Here, the effects of MRG on mammary gland differentiation and its interaction with omega-3 polyunsaturated fatty acids (omega-3 PUFAs) on growth inhibition were investigated. MRG protein expression was associated with human mammary gland differentiation, with the highest expression observed in the differentiated alveolar mammary epithelial cells from the lactating gland. Overexpression of MRG in human breast cancer cells induced differentiation with changes in cellular morphology and a significant increase in the production of lipid droplets. Treatment of mouse mammary gland in organ culture with MRG protein resulted in a differentiated morphology and stimulation of beta-casein expression. Treatment of human breast cancer cells with the omega-3 PUFA docosahexaenoic acid resulted in a differential growth inhibition proportional to their MRG expression. MRG-transfected cells or MRG protein treated cells were much more sensitive to docosahexaenoic acid-induced growth inhibition than MRG-negative or untreated control cells. Our results suggest that MRG is a candidate mediator of the differentiating effect of pregnancy on breast epithelial cells and may play a major role in omega-3 PUFA-mediated tumor suppression.

Transcriptional suppression of synuclein gamma (SNCG) expression in human breast cancer cells by the growth inhibitory cytokine oncostatin M.

Previously, we have shown that synuclein gamma (SNCG), a member of the brain protein synuclein family, is highly expressed in human infiltrating breast carcinomas but not expressed in normal or benign breast tissues. The SNCG mRNA was also detected in several human breast cancer cell lines with the highest expression found in H3922, a cell line derived from an infiltrating ductal carcinoma. In this study, we show that expression of SNCG mRNA in H3922 cells is significantly decreased by treating cells with the cytokine oncostatin M (OM) who has a growth-inhibitory effect on these cells. A decrease in SNCG mRNA level can be detected as early as 30 min after OM addition. By 4 h OM treatment, the level of SNCG mRNA was decreased to 70% of control, and by 24 h the mRNA was below detectable level. Western blot analysis further demonstrated the suppression of SNCG protein expression by OM. The level of SNCG protein in H3922 cells was reduced more than 90% by OM after 2 days. Since OM-induced growth inhibition occurs after 3-4 days, the down-regulation of SNCG expression appears to proceed the effect of OM on cell growth. Additional experiments to measure the transcriptional rates of SNCG indicate that the observed OM-induced down-regulation of SNCG mRNA occurs mainly at the transcriptional level. In an attempt to examine the role of SNCG gene in the proliferation of breast cancer cells, SNCG cDNA was stably transfected into MCF-7 cells that do not express endogenous SNCG gene. Examination of cell growth under anchorage-dependent and anchorage-independent conditions demonstrates that over expression of SNCG gene significantly stimulated the growth of MCF-7 cells both in monolayer culture and in soft agar. These data together suggest that SNCG may play a role in cell proliferation.

TIMP-4 is regulated by vascular injury in rats.

The role of basement membrane-degrading matrix metalloproteinases (MMPs) in enabling vascular smooth muscle cell migration after vascular injury has been established in several animal models. In contrast, the role of their native inhibitors, the tissue inhibitors of matrix metalloproteinases (TIMPs), has remained unproven despite frequent coregulation of MMPs and TIMPs in other disease states. We have investigated the time course of expression and localization of TIMP-4 in rat carotid arteries 6 hours, 24 hours, 3 days, 7 days, and 14 days after balloon injury by in situ hybridization, immunohistochemistry, and Western blot analysis. TIMP-4 protein was present in the adventitia of injured carotid arteries from 24 hours after injury. At 7 and 14 days after injury, widespread immunostaining for TIMP-4 was observed throughout the neointima, media, and adventitia of injured arteries. Western blot analysis confirmed the quantitative increase in TIMP-4 protein at 7 and 14 days. In situ hybridization detected increased expression of TIMP-4 as early as 24 hours after injury and a marked induction in neointimal cells 7 days after injury. We then studied the effect of TIMP-4 protein on the migration of smooth muscle cells through a matrix-coated membrane in vitro and demonstrated a 53% reduction in invasion of rat vascular smooth muscle cells. These data and the temporal relationship between the upregulation of TIMP-4, its accumulation, and the onset of collagen deposition suggest an important role for TIMP-4 in the proteolytic balance of the vasculature controlling both smooth muscle migration and collagen accumulation in the injured arterial wall.

Suppression of breast cancer growth and metastasis by a serpin myoepithelium-derived serine proteinase inhibitor expressed in the mammary myoepithelial cells.

A serpin was identified in normal mammary gland by differential cDNA sequencing. In situ hybridization has detected this serpin exclusively in the myoepithelial cells on the normal and noninvasive mammary epithelial side of the basement membrane and thus was named myoepithelium-derived serine proteinase inhibitor (MEPI). No MEPI expression was detected in the malignant breast carcinomas. MEPI encodes a 405-aa precursor, including an 18-residue secretion signal with a calculated molecular mass of 46 kDa. The predicted sequence of the new protein shares 33% sequence identity and 58% sequence similarity to plasminogen activator inhibitor (PAI)-1 and PAI-2. To determine whether MEPI can modulate the in vivo growth and progression of human breast cancers, we transfected a full-length MEPI cDNA into human breast cancer cells and studied the orthotopic growth of MEPI-transfected vs. control clones in the mammary fat pad of athymic nude mice. Overexpression of MEPI inhibited the invasion of the cells in the in vitro invasion assay. When injected orthotopically into nude mice, the primary tumor volumes, axillary lymph node metastasis, and lung metastasis were significantly inhibited in MEPI-transfected clones as compared with controls. The expression of MEPI in myoepithelial cells may prevent breast cancer malignant progression leading to metastasis.

Stimulation of breast cancer invasion and metastasis by synuclein gamma.

We recently identified and cloned novel breast cancer-specific gene BCSG1 by direct differential cDNA sequencing. BCSG1 has a great sequence homology with the Alzheimer's disease related neural protein synuclein (SNC); thus, it was also named SNC-gamma. Overexpression of SNC-gamma in breast cancer cells leads to a significant increase in motility and invasiveness in vitro and a profound augmentation of metastasis in vivo. Our data suggest that this member of the neural protein SNCs might have important functions outside the central nervous system and may play a role in breast cancer progression.

Ultrasound-guided percutaneous cholecystostomy as an initial treatment for acute cholecystitis in elderly patients.

BACKGROUND/AIMS: Acute cholecystitis may atypically present itself in the elderly, thus causing diagnostic and therapeutic problems, and it is well recognized as a high-risk condition for morbidity. The outcome has been attributed to the presence of severe co-morbid disease. In an attempt to minimize the postoperative morbidity and mortality, we performed ultrasound-guided percutaneous transhepatic cholecystostomy (PC) on elderly patients with acute cholecystitis for both initial treatment and subsequent diagnosis of their biliary tract disorders. METHODS: Those being more than 70 years old, had acute episode of cholecystitis for more than 48 h and still had positive Murphy's signs and distended gallbladders were candidates for ultrasound-guided PC. RESULTS: Forty-two elderly patients underwent ultrasound-guided PC. Once the condition of each patient showed signs of improvement and stability, cholangiography was performed via PC tube. The results of the cholangiography showed 20 patients with gallbladder stones, 16 with common bile duct stones and 6 with acalculous cholecystitis. Once stable enough, 32 patients underwent definite surgery, 18 having cholecystectomies, 14 having cholecystectomies and choledocholithotomies. The 6 patients with acalculous cholecystitis had the PC tube removed 3 weeks later, without further surgery. Two patients had gallbladder stones removed by choledochofiberscope. Two patients had common bile duct stone removed by endoscopic sphincteroplasty. Although postoperative complications occurred in 5 patients (11.9%), no instance of operative mortality was found. CONCLUSION: Our findings lead us to conclude that the use of PC in the early treatment of acute cholecystitis in elderly patients can decrease postoperative morbidity and mortality.