26 heterotrophic bacteria strains were isolated from grown abalone digestion guts and their fanning waters in Jiansheng Abalone Farm in Shangwei, Guangdong province. Analyses of extra-cellular pathogenic factors were performed and API 20 E strips were employed to identify all the isolates. Results indicated that isolates from digestion guts displayed greater ability of producing protease , amylase, gelatinase and/or hemolysis than those from farming waters, while their ability of producing lipase and phospholipase were lower than the later. Regardless of their source of origins, there were some isolates which had great abilities of producing extra-cellular products and most of them were Sphingomonas paucimobilis. Therefore, Sphingomonas paucimobilis should be considered as an opportunistic pathogen in the abalone fanning environments, while the digestion guts and fanning waters should be both regarded as the sources of harboring potential pathogens. In addition, apart from predominant strains, the roles of extra-cellular products of the bacteria community as a whole should be taken into consideration when dealing with fish diseases.

UDP glucosyltransferase (UDPGT) catalyzes the synthesis of secondary metabolites and plant hormones to regulate plant growth and development, pathogen defense and environmental adaptability. In this study 18 members of the RcUDPGT gene family were cloned from Tibetan Rhodiola crenulata and analyzed using bioinformatics. The tissue-specific expression, abiotic stresses and plant hormones (abscisic acid, auxin, methyl jasmonate) induced expression patterns were identified by real-time quantitative PCR. The bait vector of RcUDPGT (JX228125.1) was constructed to select interacting proteins from an Arabidopsis yeast library. The results of the bioinformatics analysis revealed that RcUDPGT nucleotide sequences were about 1 400 bp and encoded 452-498 amino acids. In the primary protein sequences, C-terminal sequences were more conserved compared with N-terminal regions, which held a PSPG (plant secondary product glycosyltransferase) domain. In the tertiary structures, RcUDPGTs contained a UDP sugar donor recognition binding site. In addition, all genes had multiple phosphorylation sites. The results of qRT-PCR showed that RcUDPGTs genes were expressed in root, stem and leaf. The expression levels were regulated by low temperature/ultraviolet light and various plant hormones (ABA, IAA, MeJA), but the expression patterns were quite different among them. For example, RcUDPGT6, RcUDPGT11, and RcUDPGT17 had the highest expression in leaves and were induced by all three hormones, suggesting that the functions of these genes might be to respond to environmental changes. RcUDPGT9, RcUDPGT10, RcUDPGT14 were most abundantly expressed in roots and were significantly induced by ABA and MeJA hormones, indicating that these genes may be involved in the synthesis and accumulation of salidroside. Yeast two-hybrid results showed that RcUDPGT did not exhibit autoactivation and cell toxicity, and two significant interactional genes were identified, AtKCR1 (AT1G67730.1) and AtSNL4 (AT1G70060). The AtKCR1 gene encodes a β-ketoacyl reductase (KCR) involved in synthesis of very long chain fatty acids. The AtSNL4 gene encodes a homolog of the transcriptional repressor SIN3, which could participate in the ABA hormone signaling pathway and enhance the transcriptional repression of AP2/EREBP class factors in Arabidopsis. These results suggest that the accumulation of the secondary metabolite salidroside in Rhodiola crenulata might be affected by several regulatory mechanisms. The above results may lay the foundation for understanding the adaptive mechanism of Rhodiola crenulata in a high altitude environment and stimulate an in-depth study of the synthesis and accumulation of secondary metabolites in this species.

Objective: To investigate the effects of ω-3 fish oil fat emulsion on nutritional status and humoral immunity in postoperative patients suffering from gastrointestinal malignancy. Methods: Thirty patients of gastrointestinal malignancy were randomly divided into study group (n = 15) and control group (n = 15). All the patients were assigned to receive total parenteral nutrition with the equal nitrogen and calory,and those in study group received fish oil fat emulsion additionally. Liver and renal function, blood lipid, haemoglobin, albumin, transferrin, total lymphocyte count (TLC) , B lymphocyte subsets (B1, B2), immunoglobin(IgG, IgM, IgA) and complement(C3, C4) were determined preoperatively and 1, 6d postoperatively. Results: There were no significant differences in liver and renal function and blood lipid on postoperative day 6 versus preoperation in all the two groups. TLC, IgG, IgM, C3 on postoperative day 6 were siginificantly higher in the study group(P < 0. 05). Haemoglobin, albumin, transferrin and B lymphocyte subsets were not significantly different between the two groups. Conclusion: Fish oil fat emulsion treatment was safe and tolerated, and could improve the humoral immunity in patients.

Objective To evaluate reoperations for benign bile duct strictures after a prewousRoux-en-Y hepaticojejunostomy.Methods Clinical date of 28 patients with previous reconstruction of Roux-en-Y hepaticojejunostomy for benign bile duct strictures were retrospectively analyzed.For data staftstical analysis t-test and stepwise logistic regression analysis were used.Results Reoperative surgery was performed for residual biliary stones with bile duct stricture in 10 cases(35.7%),simple anastomotic stricture of hepaticojejunostomy in 11 cases(39.3%),remained biliary stricture after initial rear in 6 cases (21.4%).anastomotic leakage with duodenal leakage in one case(3.6%).Mode of reoperation:18 cases (64.3%)underwent hepatic lobectomy with Roux-en-Y hepaticojejunostomy,liver splitting approach to Roux-en-Y hepaticojejunostomy in 5 cases(17.9%),right hemihepatectomy in one case(3.6%),resection of anastomotic stenosis involved segment and Roux-en-Y hepaticojejunostomy in one case(3.6%),abdominal drainage and duodenum fistulization and jejunum ostomy in one case(3.6%),choledocholithotomy with T-tube drainage in 2 cases(7.1%);Thirteen patients(46.4%)developed postoperative complications.Conclusion Biliary tract stenosis remains the main cause for reoperation in patients undergoing a faeled reconstruction.Wide and patent biliary tract drainage and reconstruction somenmes necessitate a hepatic lobectomy.

Objective To investigate the effects of recombinant human growth hormone (rhGH) on tumor growth and vascular endothelial growth factor (VEGF) expression of human gastric carcinoma xenografts in nude mice with different expressions of growth hormone receptor (GHR). Methods Immunocytochemical method was used to pick out one GHR-positive and one GHR-negative cell line. Then the cells were subcutaneously injected into 24 nude mice separately. The nude mice bearing two different kinds of human gastric caicinoma were equalges of body weight and tumor volume of nude mice were recorded. Serum concentrations of VEGF in peripheral blood were analyzed by ELISA. VEGF protein and mRNA expression in tumor tissue were detected by immunohistochemistry and RT-PCR, respectively. Results We chose SGC-7901 as GHR positive group, and MKN-45 as the negative one. For nude mice bearing GHR + SGC-7901 xenografts, the tumor volumes were significantly larger in rhGH groups than in control group (P ＜ 0.05), and the high-dose rhGH group revealed greater effect (P ＜ 0. 05).Body weights were not significantly different among three groups (P ＞ 0. 05). Serum VEGF concentration was (252.94 ± 15.32) ng/L in the high-dose rhGH group, which was significantly higher than that in control group [(49.94 ± 5.73) ng/L] and low-dose rhGH group [(167.60 ± 9.54) ng/L] (P ＜ 0.05). Moderate positive staining with VEGF was observed in the control group, while VEGF staining was strong in rhGH administration groups. The relative expression of VEGF mRNA for the high-dose rhGH group was 0. 6470 ± 0. 0447, which was significantly higher than that in control group (0. 3230 ± 0. 0258)and low-dose rhGH group (0. 412 ± 0. 0351)(P ＜ 0.05). While for nude mice bearing GHR-MKN-45 xenografts, the body weights of the rhGH-administrated groups were significantly higher than that of control group (P ＜ 0.05), while tumor growth, serum VEGF concentration, and the expressions of VEGF mRNA and protein in tumor tissue were not significantly different (P ＞ 0.05).Conclusions rhGH can promote tumor growth and increase the expression of VEGF in the GHR-highly-expressed SGC-7901 xenograft tumor model. However, such effects do not exist in GHR-negatively-expressed MKN-45 xenograft tumor model. The existence of GHR may be a key target where rhGH influences the secretion of VEGF.

BACKGROUND: Autophagy can regulate bone metabolism disorders in type 2 diabetes mellitus via nuclear factor KB receptor activating factor ligand, mammalian target of rapamycin, Wnt/β-catenin pathway, and p38 mitogen-activated protein kinase. OBJECTIVE: To analyze and summarize the role and possible molecular mechanism of autophagy to improve bone metabolism disorder in type 2 diabetes mellitus. METHODS: Using “autophagy; exercise; type 2 diabetes mellitus; bone metabolism” as keywords, we retrieved literature regarding autophagy for improving bone metabolism disorder in type 2 diabetes mellitus in PubMed and China Knowledge Network, and logically analyzed and summarized the included studies. RESULTS AND CONCLUSION: Autophagy can improve bone metabolism disorders in type 2 diabetes mellitus through activation of signaling pathways, such as PPAR-γ, Hedgehog, MITF, and peroxisome proliferator-activated receptors. Autophagy can up-regulate the differentiation capacity of osteoblasts, down-regulate the absorption capacity of osteoclasts in type 2 diabetes mellitus, and has an important effect on bone formation and osteocalcin mineralization.

Objective To investigate the effect of bifidobacteria on dextran sulphate sodium(DSS)- induced acute ulcerative colitis in mice.Methods Thirty BALB/C mice were randomly divided into nor- mal control group (n=10),0501 strain group (n=10) and c122 strain group (n=10).Fifty BALB/ C mice received 5% dextran sulphate sodium(DSS) for 7 days to induce ulcerative colitis.The mice were then divided to model group,negative control group(perfused with 0.9 NaCl solution ),positive control group(perfused with SASP of 20 mg/ml),DSS + 0501 strain group(perfused with 1?10~9 CFU/ml bifidobacteria 0501 strain solution and DSS + c122 strain group (perfused with 1?10~9 CFU/ml bifidobacteria c122 strain solution).All mice were sacrificed 9 days later.The colon specimens were measure by histoehemical staining with H-E.The expressions of interleukin-10 (IL-10) and its protein were detected by RT-PCR and immunohistochemistry respectively.Results The degree of colon inflam- mation in mice both in DSS+ 0501 strain and DSS+ c122 strain groups were aggravated and expressions of IL-10 mRNA and protein were reduced compared to model group.No colon inflammation was found in 0501 strain and c122 strain groups.Conclusion Some strain of bifidobaeteria may aggravate colon in- flammation in mice when mucosal harrier is destroyed.

ObjectiveTo explore clinical,histopathological and genetic features in a case of fatal familial insomnia (FFI) and related literatures were reviewed. Methods The clinical features in one patient with FFI were analyzed,and the dead patient was examined at autopsy and histopathological studies were performed on the brain tissues; and the blood samples from the patient and some of her familial members were collected for the sequencing of prion protein gene (PRNP). Results The main clinical features included intractable insomnia,psychiatric symptoms and abnormal night sleep behavior,unsteady gait,difficulty swallowing,sudden death,and positive family history. The pathological studies showed multiple neuronal loss and gliosis of brain tissues from the proband,predominated in thalamus; and analysis of PRNP revealed gene D178N mutation,and linkage with 129 methionine (Met) allele in the proband and a relative.ConclusionsFFI patients may manifest as sudden death,and may have prominent psychiatric symptoms; the corresponding gene mutation could occur in the asymptomatic carriers; the data of autopsy and brain tissue pathology is helpful for further understanding of this disease.

Objective To verify the reference interval for D-dimer assay and analyze the influence of age and gender on the ref-erence interval.Methods Inclusion criteria for reference individuals were established.60 healthy males and 63 females were enrolled and divided to three groups by age,including 20 to 39 years old group (20 males and 20 females),40 to 59 years old group (20 males and 23 females)and above 60 years old group (20 males and 20 females).Blood samples were drawn in cit-rate sodium anticoagulated tubes and D-dimer concentration was determined by three different coagulation analyzers using o-riginal reagents.According to CLSI guideline C-28-A3,the reference interval for each measurement system from reagent manufacturer was verified and the difference of D-dimer concentration between different age-group and sex-group was ana-lyzed using non-parameters tests.Results All reference intervals were verified for people under age 40,while one reference interval cannot be verified for people from 40 to 59 years old as same as one for people above 60 years old.D-dimer concen-tration increased with age and there was significantly different between 20~39 years old group and 40~59 years old group or above 60 years old group(P<0.05).There was only a significant difference between sex-group for people under age 60(P<0.05).Conclusion D-dimer concentration was associated with age and sex.For people under age 40,the reference inter-val from reagent manufacture can be verified and directly used in laboratory,while for people above age 60,the reference in-terval from reagent manufacture cannot be verified.The cause should be investigated and a new reference interval should be established separately when necessary.

Objective To compare the bone microstructure and osteoblast and osteoclast activity in different regions of osteonecrosis of the femoral head.Methods The osteonecrosis femoral heads were collected from 10 patients (Ficat Ⅳ) who had undergone total hip arthroplasty from March 2011 to May 2013.There were 6 males and 4 females.Their average age was 47.7 years old (range,40-57 years).The samples were divided into subchondral bone region,necrotic region,sclerosis region and healthy region according to radiographic results,then the bone microstructure,micro mechanism and osteoblasts/osteoclasts activity were analyzed byMicro-CT,RT-PCR,Nanoindentation,immunohistochemistry and Trap staining.Results According to the micro-CT results,the continuity of trabecular bone in necrotic region was damaged.The number of trabecular was increased and the gap was narrowed in sclerosis region.The shape and number of trabecular bone were normal in the healthy region.The elasticity moduli in different regions were:subchondral bone region 13.808±4.22 GPa,necrotic region 13.999±3.816 GPa,sclerosis region 17.266±3.533 GPa and healthy region 11.927±1.743 GPa.The hardness were subchondral bone region 0.425±0.173 GPa,necrotic region 0.331±0.173 GPa,sclerosis region 0.661±0.208 GPa,and healthy region 0.423±0.088 GPa.The trap staining of subchondral bone in healthy region and necrotic region were positive while other regions were negative.Immunohistochemistry staining showed that compared with necrotic region,the RANK and RANKL staining level increased significantly in subchondral bone and necrotic region,while Runx2 and BMP2 staining level increased significantly in sclerosis region.Conclusion The mechanical properties of trabecular have no significant difference between necrotic region and healthy region in the progress of the osteonecrosis,while the bone structure has obvious changes.An active bone resorption is observed in subchondral bone and necrotic region,while a higher bone formation activity is found in sclerosis region.