Objective To study the preventive effects of vitamin K2 on tumor recurrence in patients with hepatocellalar carcinoma (HCC) after radical resection. Methods The clinical data of 50 patients with HCC who received radical resection from March 2006 to March 2007 in No. 181 Hospital of PLA were analyzed retrospec-tively. All the patients were divided into 2 groups according to the random number table. Twenty-six patients in vitamin K2 group were administered with menatetrenone (45 mg per day), and the rest 24 pateints were in the control group. The accumulative and tumor-free survival rates, differences between the 2 groups, multivariate factors for prognosis were analyzed by Kaplan-Meier curve, Log-rank test and Cox regression model, respectively. Results During a period of 36 month follow-up, 10 patients died and 28 had tunor recurrence. The 1-, 2-, 3-year accumulative survival rates were 96%, 92% and 83% in vitamin K2 group, and 96%, 82% and 63% in control group (χ2 = 3.61, P > 0.05). The 1-, 2-, 3-year tumor-free survival rates were 92%, 60% and 38% in vitamin K2 group, and 75%, 42% and 12% in control group, with significant difference between the 2 groups (χ2 =5.61, P <0.05). Univariate and multivariate Cox proportional hazard analysis showed that without taking menate-trenone, the preoperative level of alpha fetoprotein (AFP) ≥800 μg/L and vascular invasion were the indepen-dent risk factors for tumor recurrence. Conclusions Vitamin K2 has a suppressive effect on tumor recurrence of HCC, while patients with AFP≥800 μg/L before operation or with vascular invasion have poor prognosis.

Glucose-sensitive materials have attracted much interest due to their potential application in diabetes treatment in recent years.Phenylboronic acid-based glucose-responsive polymers,which possess continuous glucose sensitivity and good stability,have been most widely used in self-regulated insulin delivery.This review covers the recent advances in PBA-functionalized nanogels (microgels),micelles,vesicles and nanoparticles.Synthesis and application of these nanomaterials are discussed.With the development of PBA-regulated polymers,these nanomaterials will migrate from laboratory to clinical use in the near future.

Objective To explore the efficacy of neoadjuvant chemoradiotherapy (CRT) followed by surgery for locally advanced esophageal squamous cell carcinoma (ESCC),and to investigate the correlation between a clinical complete response (cCR) and a pathologic complete response (pCR).Methods One hundred and fifty-eight patients with locally advanced thoracic ESCC from 2001 to 2013 were retrospectively analyzed.All patients received concurrent chemoradiotherapy followed by surgery.Platinumbased chemotherapy regimens were adopted in chemotherapy and a prescribed dose of 40 Gy in 20 fractions,5 fractions per week,was used in radiotherapy.The overall survival (OS) and disease-free survival (DFS) rates were calculated using the Kaplan-Meier method,and pairwise comparisons and univariate prognostic analyses were performed using the log-rank test.Multivariable prognostic analyses were performed using the Cox regression model.Results The pCR rate was 41.1％ in all patients.After the treatment with neoadjuvant CRT,32(72.7％) out of 44 patients with a cCR had a pCR,but only 33(28.9％) out of 114 patients with a non-cCR had a pCR (P =0.000).The sensitivity,specificity,positive predictive value,and negative predictive value of a cCR in predicting a pCR were 49.2％,87.1％,72.7％,and 71.1％,respectively.The 3-year sample size was 91.The 3-year OS and DFS rates in all patients were 53.9％ and 48.6％,respectively.Patients with a cCR had significantly higher 3-year OS and DFS rates than those with a non-cCR (P =0.012;P =0.026),while patients with a pCR had significantly higher 3-year OS and DFS rates than those with a non-pCR (P =0.000;P =0.000).The multivariate analyses demonstrated that the pathologic response after CRT and chemotherapy regimen were the influencing factors for OS.The most common grade ≥3 acute adverse reaction was leucopenia (34.2％).Conclusions With a high pCR rate and tolerable adverse reactions,neoadjuvant CRT followed by surgery is a safe and effective option for locally advanced ESCC.The cCR rate after CRT is closely correlated with the pCR and OS rates.

This study is to investigate the effect and mechanism of puerarin on DNA damage of HaCaT cells induced by UVB. Puerarin pre-treated cells were irradiated with UVB at 30 mJ x cm(-2). Twenty four hours after irradiation, DNA damage was detected by comet assay, ceramide was measured by thin layer chromatography and gas chromatography, intracellular free calcium ion was analyzed by flow cytometry, the phosphorylation level of p38 protein was examined by Western blotting method. Levels of DNA damage, ceramide, free calcium ion and p-p38 protein were elevated in UVB model cells. Contrary to the model group, all indicators above were reduced in all groups pre-treated by puerarin. Puerarin restrains the ceramide accumulation to block downstream p38 MAPK pathway and calcium ion rising, therefore reduces DNA damage in HaCaT cells induced by UVB.
Calcium ; metabolism ; Cell Line ; Ceramides ; metabolism ; DNA Damage ; drug effects ; radiation effects ; Down-Regulation ; Humans ; Isoflavones ; pharmacology ; Keratinocytes ; cytology ; metabolism ; Phosphorylation ; Signal Transduction ; drug effects ; Ultraviolet Rays ; adverse effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

The production of human recombinant proteins in milk of transgenic farm animals offers a safe, very cost-effective source of commercially important proteins that cannot be produced as efficiently in adequate quantities by other methods. This review has summarized the current status of gene selection, vector construct, transgenic methods, economics, and obvious potential in transgenic animals bioreactors. Recently, a more powerful approach was adopted in the transgenic animals founded on the application of nuclear transfer. As we will illustrate, this strategy presents a breakthrough in the overall efficiency of generating transgenic farm animals, product consistency, and time of product development. The successful adaptation of Cre-/lox P-mediated site-specific DNA recombination systems in farm animals will offer unprecedented possibilities for generating transgenic animals.
Animals ; Animals, Genetically Modified ; Bioreactors ; Breast ; metabolism ; Cell Transplantation ; Gene Expression ; Humans

OBJECTIVETo investigate the feasibility of using adipose tissue derived stem cells (ASCs) to promote neovascularization and survival rate of free fat transplantation.

METHODSASCs were isolated from aspirates from human liposuction and cultured in vitro. The cells were incubated in adipogenic, osteogenic, and chondrogenic medium for 2-4 weeks to induce adipogenesis, osteogenesis and chondrogenesis, respectively. ASCs were labelled by DiI. ASCs (A group), Insulin (B group), Medium (C group) were respectively mixed with free fat graft from aspirates. The mixtures were injected subcutaneously at the three random points on the back of eighteen 4- 6-week-old nude mice. Transplanted fat tissue was harvested after 6 months. The grafts were assessed by morphological observation, HE staining and immunohistochemistry.

RESULTSASCs can be easily harvested from liposuction aspirates and differentiate into adipogenic, osteogenic, chondrogenic lineages. The wet weight of transplanted fat tissue in ASCs group was (165.97 +/- 5.51) mg, significantly higher than that in the insulin group (93.42 +/- 5.12) mg and control group (67.64 +/- 5.09) mg (P = 0.000). The rate of fibrosis and steatonecrosis in ASCs group was( 152.2 +/- 9.8)/10HF, significantly lower than that in the Insulin group (743.9 +/- 20.4)/10HF and control group (892.2 +/- 16.5)/10HF (P = 0.000). DiI labelled ASCs were found between adipocytes and in the connective tissue in free transplanted fat tissue, and some of these cells were immunopositive for antihuman CD31 and FITC, suggesting differentiation into vascular endothelial cells.

CONCLUSIONSASCs can differentiate into vascular endothelial cells and contribute to angiogenesis in free transplanted fat tissue. ASCs can increase the survival rate and decrease the rate of fibrosis and steatonecrosis of free transplanted fat tissue. These findings suggest that ASCs-assisted transplantation may be an ideal cell therapy.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; transplantation ; Adult ; Animals ; Cell Differentiation ; Cells, Cultured ; Feasibility Studies ; Female ; Graft Survival ; Humans ; Mice ; Mice, Nude ; Neovascularization, Physiologic ; Stem Cells ; cytology ; Tissue Scaffolds

To investigate the effects of intratracheal instillation of PM2.5 suspension on bleomycin (BLM)-induced pulmonary fibrosis in mice and the intervention of neotuberostemonine (NTS), the BLM dose (1.5 or 3.0 U/kg) and PM2.5 frequency (1 or 2 times per week) were studied by factorial experiment design. After intratracheal instillation of BLM (1.5 or 3.0 U/kg) on day 0, PM2.5 (5 mg/kg) was intratracheally injected to mice once or twice a week from day 1 to day 21, and the mice in the treatment group were given 30 mg/kg NTS by gavage once a day from day 8 to day 21. The degree of pulmonary fibrosis was evaluated by lung coefficient, hydroxyproline (HYP) content, HE staining and Masson staining lung sections as well as their semi-quantitative index (HE inflammatory score and collagen volume fraction, CVF). The results showed that the HE scores increased significantly in mice singly given PM2.5 once a week, the HYP content and HE score increased in mice singly given PM2.5 twice a week, but their CVF values did not significantly increase. However, the CVF values increased significantly in mice treated with PM2.5 and BLM co-infusion. These results suggested that PM2.5 (administered singly) could significantly increase BLM-induced collagen deposition and greatly aggravate pulmonary fibrosis although it mainly caused pulmonary inflammation rather than pulmonary fibrosis. NTS could significantly reduce the CVF value and α-SMA protein level of the model mice. It can be concluded that PM2.5 has great influence on patients with respiratory diseases, while NTS can improve pulmonary fibrosis induced by the combination of PM2.5 and BLM.

OBJECTIVETo identify gene expression patterns in distinct stages of intestinal-type gastric cancer(GC).

METHODSGene expression patterns of distinct stages of intestinal-type GC samples from 3 patients were compared with cDNA microarray, which contained 576 genes. There were 506 target genes, which included 51 genes identified from our previous experiment with suppression subtractive hybridization(SSH) and other 455 genes chosen for their important roles in cancers. Hierarchical clustering was performed to clarify genes in association with distinct stages of GC.

RESULTSOne hundred and eighty-one differentially expressed genes with average Cy5:Cy3 ratios higher than 2.0 or lower than 0.5 in at least one stage of GC were identified by cDNA microarray. Among them, 48 genes were up-regulated and 133 down-regulated. Hierarchical clustering analysis separated the differentially expressed genes in different stages of GC into 5 main characteristic groups. Some important differentially expressed genes in different stages of GC were identified, such as SEC23IP, LIPF, ES(BQ291520), SLC5A1, PG(encoding similar to pepsin A precursor), CXCR4, DICER1, SH3GL2, and IGF2R.

CONCLUSIONThe differentially expressed gene patterns and some important genes were identified, which might be useful in further study on carcinogenesis, progression and metastasis of intestinal-type GC.

DNA, Neoplasm ; analysis ; Gene Expression ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Microarray Analysis ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; Stomach Neoplasms ; genetics ; metabolism ; Transcription, Genetic

This paper describes the use of piezo-driven micropipette for intracytoplasmic sperm injection of mice eggs. The head of fresh spermatozoa from KM (Kunming) fertile mice was individually injected into mature oocytes of hybrid mice B6D2F1. Approximately eighty three percent of sperm-injected oocytes survived, and 84.0% of them fertilized normally (extrusion of the second polar body and formation of male and female pronuclei). The eggs fertilized by sperm injection could develop in vitro to 2-cell (98% vs 94.7%), 4-cell (89.5% vs 92.1%) stages, no significantly (P > 0.05) different from embryos fertilized in vivo but there were significantly (P < 0.01) few morulae (63.8% vs 84.2%) and blastocysts (25.7% vs 68.4%) developed in vitro after further culture in vitro in the group of ICSI. When 120 embryos at the pronuclear stage were transferred to seven pseudopregnant KM female, 23.3% of the embryos (0 - 50%, depending on the host) reached the full term. Except for three that were cannibalized soon after birth, all of the young (25 pups) developed into normal and fertile adult. Here we report the first birth of mouse offspring following ICSI in China. These studies may increase understanding of the fertilization process and of how ICSI works.
Animals ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Male ; Mice ; Oocytes ; physiology ; Pregnancy ; Sperm Injections, Intracytoplasmic ; methods

Myostatin, a member of the TGF-beta family, negatively regulates skeletal muscle development. Mutation of myostatin activity leads to increases muscle growth and carcass lean yield. The bovine myostatin mutation cDNA was amplified by polymerase chain reaction, and then sub-cloned into the expression vector pET-30a( + ) to form the expression plasmid pET30a (+)-action/ Myostatin. The recombinant plasmid was transformed into E. coli BL21. The overexpression product of pET30a (+)-action/ Myostatin was been showed in vitro. Sheep skeletal muscle cell were cultured with the purified myostatin mutation C-terminal peptide. The results of this study suggest that had a powerful activity to stimulate the hyperplasia and proliferation of sheep muscle cells and shows high biochemical activity.
Animals ; Cattle ; Cell Proliferation ; Cells, Cultured ; Cloning, Molecular ; Genetic Vectors ; genetics ; Muscle Development ; genetics ; physiology ; Muscle, Skeletal ; cytology ; metabolism ; Mutation ; Myostatin ; genetics ; metabolism ; Peptides ; genetics ; metabolism ; Sheep