Establishment and evaluation of in vitro galectin-3 inhibitor screening model / 药学学报

Galectin-3 (Gal-3) belongs to the galectin family and is specific in binding β-galactoside. Through its C-terminal domain, Gal-3 binds to the galactoside group of the glycosylated insulin receptor (IR) and inhibits IR signaling pathway, which leads to the insulin resistance. Thus, Gal-3 is a potential therapeutic target for the treatment of insulin resistance and type 2 diabetes. Here we report a simple Gal-3 screening model based on the property that Gal-3 binds to the galactoside. We expressed and purified human Gal-3 in Escherichia coli (E.coli), and labeled it with fluorescein isothiocyanate (FITC) in vitro. After incubating FITC labeled Gal-3 (Gal-3-FITC) with PANC-1 cells, which express glycosylated membrane protein, PANC-1 cells started to show green fluorescent signal due to the Gal-3-FITC binding to the glycosylated membrane protein. Gal-3 inhibitor disrupts the binding of Gal-3-FITC and PANC1 cells, subsequently leads to the decrease of the fluorescent signal in PANC-1 cells. We can evaluate the inhibitory efficiency of Gal-3 inhibitors through measurement of the fluorescent signal. Further studies show this model is simple, stable, and repeatable with a Z' factor between 0.7 and 0.85. In sum, we have successfully established an in vitro high-throughput screening model for Gal-3 inhibitors.

Identification of Aerial Roots of Ficus microcarpa Linn. F. and Ficus elastica Roxb. Ex Hornem / 广州中医药大学学报

Objective To compare the pharmacognosy characteristics of aerial roots of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem.. Methods Fresh aerial roots were harvested and were used as the experimental samples. Stereoscopy was used for the observation of macroscopic appearance of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem.,and the microscope was used for the examination of their microscopic features of the velamen surface, cross section of root tip, cross section and longitudinal section of the posterior root, and powder. Results The appearance characteristics of the two species were as follows:the number of aerial roots of Ficus microcarpa Linn. f. was more,and the diameter was smaller than that of Ficus elastica Roxb. ex Hornem. The root tips of Ficus microcarpa Linn. f. aerial roots were light yellow turning to yellow-white, covered with gray or yellowish-white lenticels;the root tips of Ficus elastica Roxb. ex Hornem. aerial roots were light yellow or yellow, covered with gray lenticels. Microscopic identification results of the two plants were as follows:the primary xylems of transverse section of root tips and posterior roots of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem. were different,the former being five to seven heptarch,and the latter being six to eleven heptarch. Both of the two species had non-articulated unbranched laticifers in their longitudinal section of posterior root, and the diameter of Ficus. elastica Roxb. ex Hornem. was slightly larger than that of Ficus microcarpa Linn. f.. The powder of Ficus microcarpa Linn. f. was red brown,with spiral and pitted vessels;Ficus elastica Roxb. ex Hornem. was yellow brown,with single small and large pitted vessels,and the color of its fiber was shallow or nearly colorless or even transparent, with lines of cluster crystal. Conclusion The results will provide evidence for the identification , exploitation and utilization of Ficus microcarpa Linn . f . and Ficus elastica Roxb. ex Hornem.

Undergone and expecttation from Sepsis 1 to Sepsis 3 of Sepsis / 医学研究生学报

The knowledge about sepsis has undergone a transition from that of Sepsis 1.0 to that of Sepsis 3.0, which reflects a deeper insight into this syndrome -multiple organ dysfunctions resulting from SIRS-CARS, immunity, metabolism, microcirculation, and other comprehensive factors .Although sepsis 3.0 does not represent a perfect understanding of the disease , it is an improvement on the cognition of the body's response to the stimulation of infection .

Undergone and expecttation from Sepsis 1 to Sepsis 3 of Sepsis / 医学研究生学报

The knowledge about sepsis has undergone a transition from that of Sepsis 1.0 to that of Sepsis 3.0, which reflects a deeper insight into this syndrome -multiple organ dysfunctions resulting from SIRS-CARS, immunity, metabolism, microcirculation, and other comprehensive factors .Although sepsis 3.0 does not represent a perfect understanding of the disease , it is an improvement on the cognition of the body's response to the stimulation of infection .

Toll-like receptor 4/nuclear factor-kappa B signaling detected in brain after early subarachnoid hemorrhage / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Inflammation and immunity play a vital role in the pathogenesis of early brain injury after subarachnoid hemorrhage (SAH). Nuclear factor-kappa B (NF-kappaB) regulates many genes essential for inflammation and immunity and is activated by toll-like receptor (TLR). This study aimed to detect the expression of the toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-kappaB) signaling in the rat brain after early SAH.</p><p><b>METHODS</b>The rats were decapitated and their brains were removed at 0, 2, 4, 6, 12, 24 and 48 hours after a single injection of blood into the prechiasmatic cistern. mRNA expression of TLR4 was measured by Taqman real-time RT-PCR, and protein expression by immunohistochemistry and Western blotting. NF-kappaB activity and concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>TaqMan real-time RT-PCR and Western blotting identified a biphasic change in TLR4 expression in both mRNA and protein: an initial peak (2 - 6 hours) and a sustained elevation (12 - 48 hours). Immunohistochemical staining showed the inducible expression of TLR4-like immunoreactions predominantly in glial cells and vascular endothelium. A similar biphasic change in the activation of NF-kappaB subunit p65 as well as the production of NF-kappaB-regulated proinflammatory cytokines (TNF-alpha, IL-1beta and IL-6) were detected by ELISA.</p><p><b>CONCLUSIONS</b>These data suggest that experimental SAH induces significant up-regulation of TLR4 expression and the NF-kappaB signaling in early brain injury. Activation of the TLR4/NF-kappaB signaling may regulate the inflammatory responses after SAH.</p>

Cloning and expression of a new glucoamylase gene / 生物工程学报

According to the reported gene sequence of Rhizopus oryzae glucoamylases, the glucoamylase gene containing four introns was cloned from the total DNA of the natural Rhizopus arrhizu. Specific primers were designed to delete introns by overlapping PCR and a new cDNA sequence of Rhizopus arrhizu glucoamylase was obtained. The accession number in gene bank is DQ903853. This gene is successfully expressed in the Picha pastoris, producing a new protein with a high activity of glucoamylase.

Detection of methylation of hMSH2 gene promoter region of esophageal cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect methylation in promoter region of hMSH2 gene in esophageal cancer.</p><p><b>METHODS</b>Specimens of cancer and normal tissues freshly removed from 32 cases of esophageal cancer patients without previous radiotherapy, chemotherapy or other treatment were preserved at -80 degrees C within 30 min. Methylation specific PCR (MSP) was used to detect methylation of mismatch repair gene (MMR) hMSH2 in promoter region in esophageal cancer and normal esophageal tissues.</p><p><b>RESULTS</b>The frequencies of methylation of hMSH2 gene in promoter region of cancer and normal esophageal tissues were 32.4% (11/32) and 0/30 (0%), respectively, and significant difference was found between the two groups (P < 0.01). The frequency of methylation in elder patients (> or = 70 years old) was significantly higher than that in younger patients (< 70 years old) (P < 0.05). Methylation was less frequently found in grade I-II (18.2%) than in grade III-IV (70.0%) (P < 0.05).</p><p><b>CONCLUSION</b>Methylation of hMSH2 gene in promoter region is related to patients' age and histopathological grade of the esophageal cancer.</p>