[A multicenter clinical trial of collar-button type keratoprosthesis implantation for the treatment of corneal blindness in high-risk transplantation].

Objective: To evaluate the efficiency and safety of collar-button type keratoprosthesis (c-bKPro) implantation for corneal blindness in high-risk transplantation in China. Methods: It was a case series study. High-risk corneal blind patients who planned to undergo c-bKPro implantation were prospectively and continuously enrolled in the Eye Hospital of Shandong First Medical University, Ophthalmology Division of Chinese PLA General Hospital, Zhongshan Ophthalmic Center, Department of Ophthalmology in Eye & ENT Hospital of Fudan University, and Eye Hospital of Wenzhou Medical University from July 2019 to January 2020. The cure for blindness and surgical success were assessed based on visual acuity (VA)≥0.05. The complications and keratoprosthesis retention rate were recorded to determine the safety of the surgery. Results: Thirty-seven subjects (eyes) were included, of which 32 were male and 5 were female, aged 27 to 72 years old. The indications of c-bKPro implantation were corneal graft failure (21 eyes, 56.8%), chemical injury (8 eyes, 21.6%), thermal burn (5 eyes, 13.5%), unexplained corneal opacity (2 eyes, 5.4%), and corneal perforation (1 eye, 2.7%). Two patients withdrew from the clinical trial at 3 months postoperatively. Thirty-five patients were followed up for 6 months, and 31 were followed up for 12 months. The VA was ≥0.05 in 83.8% of eyes at 6 months and in 81.8% of eyes at 12 months. Among the 11 eyes diagnosed with concurrent glaucoma, 6 eyes achieved a VA of ≥0.05. At 12 months, the c-bKPro retention rate was 100%. The surgical complications included retroprosthetic membrane formation (5 eyes, 16.1%), persistent corneal epithelial defects (5 eyes, 16.1%), macular edema (4 eyes, 12.9%), new-onset glaucoma (4 eyes, 12.5%; including one eye withdrawn from the study at 3 months), sterile corneal melting (2 eyes, 6.5%), sterile vitritis (1 eye, 3.2%), and infectious keratitis (1 eye, 3.2%). Conclusions: C-bKPro implantation is an effective and safe option for treating corneal blindness in high-risk transplantation in China. Improved visual outcomes could be achieved in most cases, with a relatively low incidence of postoperative complications.

TNFAIP3 inhibits migration and invasion in nasopharyngeal carcinoma by suppressing epithelial mesenchymal transition.

TNF alpha induced protein 3 (TNFAIP3), a member of zinc finger protein family, is a gene whose expression level is promptly induced by the tumor necrosis factor. In this study, the clinical significance of TNFAIP3 was analyzed based on available samples in The Cancer Genome Atlas database. TNFAIP3 downregulation was associated with distant metastasis and worse patient prognosis. TNFAIP3-overexpressing and TNFAIP3-knockdown NPC cell line models were established through plasmid-mediated overexpression and small interfering RNA (siRNA), respectively. Cell migration and invasion capacities were evaluated by wound-healing and transwell assays. Functional studies indicated that TNFAIP3 knockdown promoted migration and invasion, whereas TNFAIP3 overexpression alleviated these functions. Western blot analysis was used to examine protein changes from TNFAIP3 overexpression and knockdown, in which TNFAIP3 promoted the protein expression of E-cadherin and suppressed vimentin expression. Our data suggested that TNFAIP3 inhibited migration and invasion by suppressing epithelial mesenchymal transition in NPC.

[Dynamic expression of carnitine palmitoyltransferase II in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation].

Objective: To investigate the dynamic expression of hepatic carnitine palmitoyltransferase-II (CPT-II) in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation. Methods: Male Sprague-Dawley rats were divided randomly into control, fatty liver, and induced cancer groups, which were fed with normal, high-fat (HF), and HF containing 2-fluorenylacetamide (0.05%, 2-FAA) diets, respectively, for 14 weeks. One rat from each group was sacrificed every two weeks and the blood and liver samples were collected. Liver morphological changes were evaluated with hematoxylin and eosin staining, and the liver tissue samples were divided into control, fatty liver, degeneration, precancerous, and cancerous groups accordingly. Hepatic lipids were dyed by the oil red O method. The CPT-II expression was measured by immunohistochemistry and compared with the specific CPT-II concentration (ng/mg liver protein, ng/mg P) among different groups. Serum levels of circulating total cholesterol (Tch), triglyceride (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively analyzed. Results: Massive lipid accumulation hepatocytes was seen in rats on HF and HF containing 2-FAA diets. The lipid levels in the control group were significantly lower than those in the fatty liver (t = -11.556, P < 0.001), degeneration (t = -4.847, P = 0.04), precancerous (t = -13.652, P = 0.005), and cancerous groups (t = -10.896, P = 0.008). The serum TG and Tch levels in the degeneration, precancerous, and cancerous groups were 2-3 times higher than those in the control group (P < 0.05). After 2-FAA treatment, the morphological changes of rat hepatocytes showed the progression from degeneration and precancerosis to cancerosis, with hepatocyte injury. The serum AST and ALT levels in the degeneration, precancerous, and cancerous groups were significantly higher (4-8 times) than those in the control group (P < 0.05). The specific concentration of liver CPT-II expression was significantly reduced during hepatocyte malignant transformation, as confirmed by immunohistochemistry, with the CPT-II levels significantly lower in the cancerous group than in any of other groups (P < 0.05). Conclusion: Low hepatic CPT-II expression might lead to abnormal lipid accumulation in hepatocytes, which should promote the malignant transformation of hepatocytes.

[Role of inhibition of nuclear factor-kappa B gene transcription by specific miRNA in reversing multi-drug resistance of liver cancer].

Objective: To investigate the reversal effect of inhibition of nuclear factor-kappa B (NF-κB) gene transcription by specific miRNA on multi-drug resistance (MDR)of liver cancer. Methods: The expression of P-glycoprotein (P-gp) and NF-κB in hepatoma cells, drug-resistant HepG2/ADM cells, and liver cells (LO2 cells) was analyzed. Specific NF-κB miRNA plasmids were constructed, screened, and transfected into HepG2 or HepG2/ADM cells. Western blot was used to measure the concentrations of P-gp and NF-κB, and FQ-PCR was used to measure gene expression; Cell Counting Kit-8 assay was used to measure cell proliferation and the influence of drugs on cell proliferation; flow cytometry and Annexin-V-PE/7-ADD double staining were used to observe cell cycle and apoptosis. The t-test was used to compare means between groups, and a one-way analysis of variance was used to compare means between multiple groups. Results: After being treated by adriamycin, hepatoma cells showed increased expression of P-gp and an increased level of NF-κB phosphorylation. At 24, 48, and 72 hours, the resistance index of the HepG2/ADM cells (IC50 = 4.166, 1.522, and 1.380 µmol/L) was 8.519, 6.874, and 6.166 times that of the HepG2 cells (IC50 = 0.489, 0.221, and 0.224 µmol/L). The HepG2/ADM cells showed significantly higher relative mRNA expression (∆ct value) of mdr1 and NF-κB than the HepG2 cells (3.310±0.154/2.580±0.040 vs 0.084±0.038/0.6067±0.032, both P < 0.01). After being transfected with miRNA1, the HepG2/ADM cells showed significantly lower mRNA expression of mdr1 than the cells in the miRNA-negative group (2-∆∆ct = 0.326±0.011 vs 0.804±0.057, t = 14.262, P < 0.01), as well as significant reductions in the expression of intracellular t-p65, nuclear p-p65, and P-gp compared with the cells in the miRNA-negative group (P < 0.01), with inhibited cell proliferation, G1 phase arrest, and increased apoptosis. Conclusion: Abnormal expression of MDR1/P-gp is closely associated with MDR, and inhibition of NF-κB activation by specific miRNA can significantly inhibit MDR1/P-gp gene transcription and reverse MDR of liver cancer.

[Abnormal expression of insulin-like growth factor-I receptor and inhibitory effect of its transcription intervention on nude mice xenograft tumor].

Objective: To investigate the expression of insulin-like growth factor-I receptor (IGF-IR) in liver cancer and the inhibitory effect of its transcription intervention on nude mice xenograft tumor. Methods: A total of 40 patients with primary liver cancer were enrolled, and 40 samples of cancer lesions, peri-cancerous tissues (with a distance of 2 cm to the margin of cancer lesion), or distal liver tissues (with a distance of 5 cm to the margin of cancer lesion), with a weight of 200 mg, were collected after surgery. Some of these samples were used for pathological examination, and the rest were stored at -85°C. A total of 18 BALB/c nude mice aged 4-6 weeks with a body weight of 18-20 g (9 male and 9 female mice) were randomly divided into control group, negative control group, and co-intervention group, with 6 mice in each group, and fed under specific pathogen-free conditions. The cell line was cultured in the dimethyl sulfoxide complete medium containing 10% fetal bovine serum in a CO2incubator at 37°C. When the cell confluence reached 90% after cell inoculation, shRNA was divided into co-intervention group, negative control group, and untreated control group and were transfected to hepatoma cells using PolyJetTM transfection reagent. Stable cell clones obtained by G418 screening and used for the in vivo study. Immunohistochemistry, Western blotting, and quantitative real-time PCR were used to analyze the expression of IGF-IR in the human hepatoma tissue and cell line. The IGF-IR shRNA eukaryotic expression plasmids were established and screened for the most effective sequence; they were transfected to PLC/PRF/5 hepatoma cells, and the CCK-8 assay was used to analyze the changes in cell proliferation. The stable cell line screened out by G418 was inoculated to establish the subcutaneous xenograft tumor in nude mice. The tumor growth curve was plotted and histological examination was performed. Graphpad Prism 5.0 and SPSS 18.0 were used for plotting and data analysis; the variance test and Q test were used for comparison of means between multiple samples, the t-test was used for comparison of means between any two samples, the chi-square test or Fisher's exact test was used for comparison of rates between samples, and a rank correlation analysis was performed for expression intensity. Results: The liver cancer group had a significantly higher positive rate of IGF-IR than the peri-cancerous group and distal tissue group (82.5% vs 42.5%/10%,χ2= 13.653 and 42.29, bothP< 0.01), as well as significantly higher expression intensity than these two groups (Z= 4.771 and 6.579, bothP< 0.01). IGF-IR was not significantly expressed in the L02 cell line and was strongly expressed in the PLC/PRF/5 hepatoma cells, and the expression intensity of IGF-IR in the PLC/PRF/5 hepatoma cells was 4 and 5 times that in Bel-7404 cells and HepG2 cells, respectively. After the PLC/PRF/5 hepatoma cells were transfected with shRNA4 with the best co-intervention effect, the mean inhibition rate of tumor cell growth reached 63.9% at 72 hours, and the mean inhibition rate of IGF-IR transcription reached 59.6%. Tumor cells were arrested in G1 phase, and there was a significant increase in apoptosis rate. As for the subcutaneous hepatoma xenograft in nude mice, the intervention group had significantly slower tumor growth than the blank control group and negative control group (143±24 mm3 vs 372±46 mm3/350±50 mm3,t= 10.776 and 9.142, bothP< 0.01); the intervention group had significantly downregulated IGF-IR expression, which was significantly lower than that in the blank control group and negative control group (t= 11.184 and 9.450, bothP< 0.01). Conclusion: Intervention of IGF-IR transcription can effectively inhibit the growth of xenograft tumor in nude mice, suggesting that IGF-IR gene might become a new potential target for the treatment of liver cancer.

Interval multistage joint-probabilistic integer programming approach for water resources allocation and management.

In this study, an interval multistage joint-probabilistic integer programming method was developed to address certain problems in water resource regulation. This method effectively deals with data in the form of intervals and probability distribution. It can also process uncertain data in the form of joint probabilities. The proposed method can also reflect the linkage and dynamic variability between particular stages in multi-stage planning. Sensitivity analysis on moderate violations and security constraints showed that the degree of constraint violation was closely linked to the final benefits of the system. The developed method was applied in the case study of the joint-operation of the Tianzhuang and Bashan Reservoirs in Huaihe River, China. In this case study, the proposed method can deal with the water shortage problems downstream and the distribution problems caused by excess water in the reservoir. It can also guarantee the optimization of long-term water usage of both Reservoirs and the river downstream.

Ciliary neurotropic factor, interleukin 11, leukemia inhibitory factor, and oncostatin M are growth factors for human myeloma cell lines using the interleukin 6 signal transducer gp130.

Interleukin 6 (IL-6) is a major growth factor for tumor plasma cells involved in human multiple myeloma (MM). In particular, human myeloma cell lines (HMCL), whose growth is completely dependent on addition of exogenous IL-6, can be obtained reproducibly from every patient with terminal disease. Four cytokines, ciliary neurotropic factor (CNTF), IL-11, leukemia inhibitory factor (LIF), and oncostatin M (OM), use the same transducer chain (signal transducer gp130) as IL-6 and share numerous biological activities with this IL. We found that these four cytokines stimulated proliferation and supported the long-term growth of two out of four IL-6-dependent HMCL obtained in our laboratory. Half-maximal proliferation was obtained with cytokine concentrations ranging from 0.4 to 1.2 ng/ml for IL-11, LIF, and OM. CNTF worked at high concentrations only (90 ng/ml), but addition of soluble CNTF receptor increased sensitivity to CNTF 30-fold. The growth-promoting effect of these four cytokines was abrogated by anti-gp130 antibodies, contrary to results for anti-IL-6 receptor or anti-IL-6 antibodies. No detectable changes in the morphology and phenotype were found when myeloma cells were cultured with one of these four cytokines instead of IL-6. Concordant with their IL-6-dependent growth, the four HMCL expressed membrane IL-6R and gp130 detected by FACS analysis. LIF-binding chain gene (LIFR) was expressed only in the two HMCL responsive to LIF and OM.

Exosome-mediated miR-106a-3p derived from ox-LDL exposed macrophages accelerated cell proliferation and repressed cell apoptosis of human vascular smooth muscle cells.

OBJECTIVE: Atherosclerosis (AS) is a leading disease with high mortality and morbidity in the world. It has been demonstrated that exosomes can transfer some miRNAs or proteins to regulate the biological functions of human vascular smooth muscle cells (VSMCs) and promote the progression of AS. In this study, we mainly aimed at exploring potential functions of exosomes derived from ox-LDL exposed macrophages and investigating the potential mechanisms of exosome-mediated miR-106a-3p in regulating VSMCs and promoting AS. MATERIALS AND METHODS: Ox-LDL was used to treat THP-1 macrophages, CCK-8 assay was performed to detect cell viability, and flow cytometric analysis was used to detect cell apoptosis. Exosomes were isolated and collected with centrifugation, and were determined by transmission electron microscopy and WB assay. RT-PCR was used to detect the expressions of miRNAs in exosomes and VSMCs, WB assay was used to detect protein expressions. MiR-106a-3p mimic was transfected into VSMCs to verify its functions and the Luciferase gene reporter assay was performed to prove the binding site of miR-106a-3p and CASP9. Finally, GW4869, an inhibitor for exosome secretion, was used to block exosome secretion by ox-LDL induced THP-1 and to confirm the effects of miR-106a-3p on cell proliferation and apoptosis in VSMCs. RESULTS: We found that ox-LDL induced THP-1 could promote cell proliferation and repress cell apoptosis of VSMCs, then, exosomes were successfully isolated, which could promote cell proliferation and repressed cell apoptosis of VSMCs after adding into VSMCs. Furthermore, we found that miR-106a-3p was significantly increased in exosomes from ox-LDL induced THP-1 and its expression was also increased in VSMCs after adding into VSMCs. Moreover, miR-106a-3p overexpression could promote cell viability and repress cell apoptosis, as well as regulate associated protein expressions. Additionally, the Luciferase gene reporter assay confirmed that miR-106a-3p could directly bind with CASP9 and regulate Caspase signaling in VSMCs. Finally, blocking exosomes from ox-LDL induced THP-1 reduced the cell viability and promoted cell apoptosis in VSMCs. CONCLUSIONS: Above all, this study demonstrated that miR-106a-3p was increased in exosomes from ox-LDL induced THP-1 and it could promote cell proliferation and repress cell apoptosis of VSMCs. We found that the exosomes-mediated miR-106a-3p could directly bind with CASP9 and repress Caspase signaling pathway in VSMCs, which might provide a potential target for treating AS.

Study of EGCG induced apoptosis in lung cancer cells by inhibiting PI3K/Akt signaling pathway.

OBJECTIVE: To investigate the role of phosphatidylinositol-3-kinase protein kinase B (PI3K/Akt) signaling pathway in the apoptosis of H1299 lung cancer cells induced by epigallocatechin gallate (EGCG). MATERIALS AND METHODS: H1299 lung cancer cells were treated with EGCG at a dose of 10 µM, 20 µM, and 40 µM, respectively. Cell culture was performed for 72 h and then: 1, cell proliferation was detected by MTT assay; 2, cell apoptosis rate was detected by flow cytometry; 3, expression of Caspase-3, Bax, and Bcl-2 was detected by Western blot; 4, expression of PI3K, p-PI3K, Akt, and p-Akt was detected by Western blot. RESULTS: The proliferation of H1299 cells was significantly inhibited 72 h after treatment with different doses of EGCG, and cell apoptosis rate was significantly increased (p<0.05). Compared with those in the control group, expression of PI3K and Akt in the lung cancer cells H1299 after EGCG treatment showed no significant differences (p>0.05), while expression levels of p-PI3K and p-Akt were significantly reduced (p<0.05). CONCLUSIONS: EGCG can inhibit the proliferation and induce apoptosis of H1299 lung cancer cells, and the effect is related to the inhibition of the activation of PI3K/Akt signaling pathway.

Inhibition of T lymphocyte activation in mice heterozygous for loss of the IMPDH II gene.

Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in the de novo synthesis of guanine nucleotides, which are also synthesized from guanine by a salvage reaction catalyzed by the X chromosome-linked enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT). Since inhibitors of IMPDH are in clinical use as immunosuppressive agents, we have examined the consequences of knocking out the IMPDH type II enzyme by gene targeting in a mouse model. Loss of both alleles of the gene encoding this enzyme results in very early embryonic lethality despite the presence of IMPDH type I and HPRT activities. Lymphocytes from IMPDH II(+/-) heterozygous mice are normal with respect to subpopulation distribution and respond normally to a variety of mitogenic stimuli. However, mice with an IMPDH II(+/-), HPRT(-/o) genotype demonstrate significantly decreased lymphocyte responsiveness to stimulation with anti-CD3 and anti-CD28 antibodies and show a 30% mean reduction in GTP levels in lymphocytes activated by these antibodies. Furthermore, the cytolytic activity of their T cells against allogeneic target cells is significantly impaired. These results demonstrate that a moderate decrease in the ability of murine lymphocytes to synthesize guanine nucleotides during stimulation results in significant impairment in T-cell activation and function.

Synthesis, crystal structure, and anti-breast cancer activity of a novel metal-porphyrinic complex [YK(TCPP)(OH)2·(solvents)x].

A novel heterometallic metal-porphyrinic framework (MPFs) built from Y and K ions as nods and meso-tetra(4-carboxyphenyl)porphyrin as linkers has been successfully synthesized and characterized. The single crystal X-ray diffraction indicated that this complex 1 exhibited a bilayered architecture of the porphyrins, which is seldom seen in MPFs. In addition, in vitro anticancer activity of complex 1 on three human breast cancer cells (BT474, SKBr-3 and ZR-75-30) was further determined.

Inosine-5'-monophosphate dehydrogenase: regulation of expression and role in cellular proliferation and T lymphocyte activation.

Guanine nucleotide synthesis is essential for the maintenance of normal cell growth and function, as well as for cellular transformation and immune responses. The expression of two genes encoding human inosine-5'-monophosphate dehyrogenase (IMPDH) type I and type II results in the translation of catalytically indistinguishable enzymes that control the rate-limiting step in the de novo synthesis of guanine nucleotides. Cellular IMPDH activity is increased more than 10-fold in activated peripheral blood T lymphocytes and is attributable to the increased expression of both the type I and type II enzymes. In contrast, abrogation of cellular IMPDH activity by selective inhibitors prevents T lymphocyte activation and establishes a requirement for elevated IMPDH activity in T lymphocytic responses. In order to assess the molecular mechanisms governing the expression of the IMPDH type I and type II genes in resting and activated peripheral blood T lymphocytes, we have cloned the human IMPDH type I and type II genes and characterized their genomic organization and their respective 5'-flanking regions. Both genes contain 14 highly conserved exons that vary in size from 49 to 207 base pairs. However, the intron structures are completely divergent, resulting in disparities in gene length (18 kilobases for type I and 5.8 kilobases for type II). In addition, the 5'-regulatory sequences are highly divergent; expression of the IMPDH type I gene is controlled by three distinct promoters in a tissue specific manner while the type II gene is regulated by a single promoter and closely flanked in the 5' region by a gene of unknown function. The conservation of the IMPDH type I and type II coding sequence in the presence of highly divergent 5'-regulatory sequences points to a multifactorial control of enzyme expression and suggests that tissue-specific and/or developmentally specific regulation of expression may be important. Delineation of these regulatory mechanisms will aid in the elucidation of the signaling events that ultimately lead to the synthesis of guanine nucleotides required for cellular entry into S phase and the initiation of DNA replication.

Differential actions of neurotrophins on apoptosis mediated by the low affinity neurotrophin receptor p75NTR in immortalised neuronal cell lines.

The low affinity neurotrophin receptor (p75NTR) mediates apoptosis of a number of neuronal and non-neuronal cells but the signals leading to the apoptosis remain obscure. To reveal the mechanism of p75NTR-mediated apoptosis, a neural cell line expressing human p75NTR was established. The human cDNA fragment encoding for p75NTR was PCR-amplified, cloned into the retrovirus expression vector pXT-1 and transfected into the rat cerebellum cell line R2. The expression of p75NTR in the R2 cell line was demonstrated by both Northern blotting analysis and immunocytochemistry. Serum withdrawal induced dramatic apoptosis in p75NTR-expressing R2 cells (R2L1) but not in pXT-1 transfected control R2 cells (R2P). Reverse transcription polymerase chain reaction (RT-PCR) revealed that these cell lines express trkA and trkB but not trkC. The apoptosis of R2L1 cells triggered by the serum deprivation for 48 h was completely prevented by neurotrophin-3 and the antibody to p75NTR but only partially prevented by the nerve growth factor and brain derived neurotrophic factor. We conclude that the p75NTR mediates apoptosis of R2L1 cells by its intrinsic receptor effects requiring an unbound status of this receptor and that the apoptosis is prevented by neurotrophins or the antibody to p75NTR through distinct mechanisms.

Inosine monophosphate dehydrogenase expression: transcriptional regulation of the type I and type II genes.

Inosine 5'-monophosphate dehydrogenase (IMPDH) is an essential rate-limiting enzyme in the de novo guanine nucleotide synthetic pathway that catalyzes the conversion of IMP to XMP. Enzyme activity is accounted for by the expression of two distinct but closely related genes termed IMPDH I and II. Increased IMPDH activity has been linked to both cellular proliferation and neoplastic transformation and generally ascribed to an increase in the expression of the type II gene. We have characterized the type I and type II genes and identified elements important in the transcriptional regulation of both genes. The type II IMPDH gene contains a 466 bp 5' flanking region spanning the translation start site that contains several transcription factor binding sites and mediates increased transcription of a CAT reporter gene in peripheral blood T lymphocytes when these cells are induced to proliferate. The single functional IMPDH type I gene contains exon-intron boundaries and exon structures that are nearly identical to those in the type II gene. In contrast to the type II gene, however, it contains two putative promoter sites, each with the potential for transcriptional regulation. We conclude that these two genes most probably arose from an early gene duplication event and that their highly conserved structures and differential regulation at the transcriptional level argue strongly for a significant role for each gene in cellular metabolism, growth, and differentiation.

Synthesis, crystal structure, and anti-breast cancer activity of a novel metal-porphyrinic complex [YK(TCPP)(OH)2·(solvents)x]

A novel heterometallic metal-porphyrinic framework (MPFs) built from Y and K ions as nods and meso-tetra(4-carboxyphenyl)porphyrin as linkers has been successfully synthesized and characterized. The single crystal X-ray diffraction indicated that this complex 1 exhibited a bilayered architecture of the porphyrins, which is seldom seen in MPFs. In addition, in vitro anticancer activity of complex 1 on three human breast cancer cells (BT474, SKBr-3 and ZR-75-30) was further determined.

A preliminary investigation of the mechanisms underlying the effect of berberine in preventing high-fat diet-induced insulin resistance in rats.

Berberine exerts insulin resistance-improving effects, the underlying mechanism of which is not well understood. We herein aimed to examine the effects of berberine on mediators of insulin signaling in pancreatic ß- and α- islet cells and hepatocytes using a rat obesity model. Rats were fed the following diets for 22 weeks: normal control (NC); normal+berberine (NC+BBR 200 mg/kg/day); high-fat (HF); HF+BBR(1) (BBR 100 mg/kg/day); HF+BBR(2) (BBR 200 mg/kg/day). Metabolic parameters were assessed and mediators of insulin signaling were quantified by immunohistochemistry. The HF diet significantly increased body weight (BW), visceral fat (VF), the visceral fat to BW ratio (VF/BW), and insulin resistance index in the HF group compared with the NC group. Both doses of BBR significantly reduced HF diet-induced increases in BW, VF, and VF/BW. IR and IRS-1 expression in ß-cells was significantly lower in the HF group, but not the HF+BBR groups, compared with the NC and NC+BBR groups. Glucagon expression in α-cells was significantly higher in the HF group compared with all other groups. IR expression in α-cells was significantly lower in the HF group compared with the NC, NC+BBR, and HF+BBR(2) groups. IR expression in hepatocytes was significantly lower in the HF group compared with all groups. Our preliminary findings suggest that berberine may ameliorate the development of insulin resistance by differentially preventing alterations in expression of IR, IRS-1, and glucagon in ß-cells, α-cells, and hepatocytes.

Anthropometric measurements for prediction of metabolic risk among Chinese adults in Pudong new area of Shanghai.

BACKGROUND: Numerous studies have demonstrated the ability of body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR) and waist-to-height ratio (WHtR) to predict the risk of type 2 diabetes mellitus (T2DM) and cardiovascular diseases (CVD). This study aimed to evaluate the predictive value of these anthropometries for metabolic abnormalities and related diseases in Chinese adults. MATERIAL & METHODS: A cross-sectional study was conducted in 2 477 men and 3 107 women at 20-79 years old who were randomly selected from Pudong New Area of Shanghai, China, through a multistage sampling process. Anthropometric variables and blood pressure were measured according to a standardized protocol, and a fasting blood sample was collected from each subject for biochemical analysis. RESULTS: Prevalence of the metabolic syndrome was observed to increase with increasing BMI, WC, WHR and WHtR in both sexes. Participants with any metabolic abnormality had a higher body size than those without. The associations of anthropometries with each metabolic factor were significant and equal for BMI, WC, WHR and WHtR. Areas under the receiver operating characteristic curves (AUC) ranged from 0.59 to 0.72 across the 4 anthropometries in predicting individual and clusters of metabolic factors. However, none of the 4 anthropometries identified newly-diagnosed T2DM or hypertension with a high sensitivity or specificity. CONCLUSION: Our findings suggest that the independent use of BMI, WC, WHR, or WHtR may not be an effective tool to predict metabolic factors and related chronic diseases in Chinese adults.

PIM1 phosphorylates and negatively regulates ASK1-mediated apoptosis.

The serine/threonine kinase, PIM1, is involved in promoting cell survival in part by phosphorylation and inhibition of proapoptotic proteins. Apoptosis signaling kinase 1 (ASK1), a mitogen-activated protein kinase kinase kinase, is involved in the so-called stress-activated pathways that contribute to apoptotic cell death. Here we show that PIM1 phosphorylates ASK1 specifically on serine residue 83 (Ser83) both in vitro and in vivo and that PIM1 binds to ASK1 in cells by co-immunoprecipitation. Using H1299 cells, our results further demonstrate that PIM1 phosphorylation of ASK1 decreases its kinase activity induced by oxidative stress. PIM1 phosphorylation of ASK1 on Ser83 inhibited ASK1-mediated c-Jun N-terminal kinase phosphorylation as well as p38 kinase phosphorylation. Under H(2)O(2)-induced stress conditions that normally lead to apoptosis, these phosphorylation events were associated with inhibition of caspase-3 activation and resulted in reduced cell death. Moreover, knockdown of PIM1 in H1299 cells decreased phosphorylation of endogenous Ser83 of ASK1 and was associated with a decrease in cell viability after H(2)O(2) treatment. Taken together, these data reveal a novel mechanism by which PIM1 promotes cell survival that involves negative regulation of the stress-activated kinase, ASK1.

Inducible functions in hybrids of a Lyt-2+ BW5147 transfectant and the 2C CTL line.

Cytolytic activity and release of interleukin 2 (IL-2) were induced in Lyt-2-positive T-T cell hybrids by incubation with either concanavalin A or irradiated stimulator cells. Since hybrids of Lyt-2-positive class I-specific cytotoxic T lymphocytes (CTLs) with the fusable mouse thymoma cell line, BW5147, are invariably Lyt-2-negative, a derivative of BW5147 was produced by transfection which constitutively expresses surface Lyt-2.1. This cell line, 3B2, was fused with the H-2Ld-specific long term CTL line, 2C. Such hybrids expressed the transfected Lyt-2 gene but not the endogenous gene of the 2C fusion partner. That Lyt-2 plays a functional role in hybrids of 3B2 with 2C is shown by the observations that: 1) cytolysis by Lyt-2-positive hybrids was inhibited by Lyt-2-specific monoclonal antibody (mAb); 2) Lyt-2-positive but not Lyt-2-negative subclones of one such line develop specific cytotoxicity when incubated with stimulator cells; 3) Less IL-2 was released from Lyt-2-negative subclones incubated with stimulator cells than from Lyt-2-positive subclones; 4) Lyt-2-specific mAb inhibits release of IL-2 from Lyt-2-positive hybrids incubated with stimulator cells. All Lyt-2-positive hybrids expressed functional surface Lyt-3 encoded by the CTL fusion partner, demonstrating that expression of the Lyt-3 gene is not sensitive to the negative regulation which shuts off the endogenous Lyt-2 gene in hybrids of class I-specific CTLs with the 3B2 or BW5147 cell lines. The existence of inducible T-T cell hybrids expressing functional Lyt-2 and Lyt-3 provides a system for evaluation of the role(s) of Lyt-2 and Lyt-3 in the induction of function independent of cell growth.

[The clinical observation of tetracycline stained teeth covered by composite resin veneer induced of secondary gingivitis]

Using gingival index,sulcus bleeding index and plaque index of 765 tetracycline stained teeth covered by composite resin veneer after 1/2-3 years were observed.It was found that the incidence of gingivitis was over 60%.The reexamination after three months later showed that the three indexes were significantly improved by grinding teeth proper and teaching standard brushing method.