[Construction of a Mouse Model for Myeloproliferative Neoplasms and an Evaluation System].

OBJECTIVE: To construct a myeloproliferative neoplasms (MPN) transplanted mouse model with JAK2-V617F, MPLW515L or CALR-Type I gene mutation, and establish a systematic evaluation system to verify the success of model construction. METHODS: The bone marrow c-kit+ cells of the mice were obtained by the following steps: The mice were killed by cervical dislocation, the femur, tibia and ilium were separated, and the bone marrow cells were collected. The c-kit+ cells were sorted after incubation with CD117 magnetic beads. The method of constructing mouse primary mutant cells is as follows: A gene mutation vector with a GFP tag was constructed by the retroviral system, and the retroviral vector was packaged into the Platinum-E cells to obtain the virus supernatant, and then used it to infect the c-kit+ cells of mice. The MPN mouse model was constructed as follows: the mouse primary c-kit+ cells containing the mutant genes were collected after infection, and then transplanted them via the tail vein into the female recipient mice of the same species which were irradiated with a lethal dose of gamma rays (8.0 Gy). The MPN mouse model was evaluated as follows: After transplantation, the peripheral blood of the mice was regularly collected from the tail vein to perform the complete blood count test, and the size of spleen and the degree of bone marrow fibrosis were estimated. RESULTS: The mouse c-kit+ cells with the mutant genes were successfully obtained from the bone marrow. MPN mouse model was successfully constructed: The peripheral blood cells of the MPN-transplanted mice carried exogenous implanted GFP-positive cells, and the white blood cells (WBC), platelet (PLT) and hematocrit (HCT) were all increased; the body weight loss, and the water and food intake were reduced in the transplanted mice; further pathological analysis showed that the transplanted mice displayed splenomegaly and bone marrow fibrosis. These results suggested that the MPN mouse model was successfully constructed. According to the common and different characteristics of the three MPN mouse model, a preliminary evaluation system for judging the success of MPN mouse model construction was summarized, which mainly included the following indicators, for example, the proportion of GFP-positive cells in the peripheral blood of mice; WBC, PLT and HCT; the degree of spleen enlargement and the bone marrow fibrosis. CONCLUSION: The MPN mouse model with JAK2-V617F, MPLW515L or CALR-Type I gene mutation is successfully established by retroviral system, which can provide an important experimental animal model for the research of MPN pathogenesis and drug-targeted therapy.

Diabetes influences the fusion of autophagosomes with lysosomes in SH-SY5Y cells and induces Aß deposition and cognitive dysfunction in STZ-induced diabetic rats.

Diabetes has been regarded as an independent risk factor for Alzheimer's disease (AD). Our previous study found that diabetes activated autophagy, but lysosome function was impaired. Autophagy-lysosome dysfunction may be involved in Aß deposition in diabetic cognitive impairment. In the present study, we used STZ-induced diabetic rats and SH-SY5Y cells to investigate whether diabetes inhibits autophagosome fusion with lysosomes. We found that in the in vivo study, STZ-induced diabetic rats exhibited cognitive dysfunction, and the lysosome function-related factors CTSL, CTSD, and Rab7 were decreased (P < 0.05). In an in vitro study, the mRFP-GFP-LC3 assay showed that the fusion of autophagosomes with lysosomes was partly blocked in SH-SY5Y cells. High glucose treatment downregulated the number of autophagolysosomes, downregulated CTSD, CTSL, and Rab7 expression (P < 0.05), and then influenced the function of ACP2 to partly block the fusion of autophagosomes and lysosomes to inhibit Aß clearance. These findings indicate that high glucose treatment affected lysosome function, interfered with the fusion of autophagosomes with lysosomes, and partly blocked autophagic flux to influence Aß clearance.

Coexisting cancers: a mixture of neuroendocrine carcinoma and adenocarcinoma in the gallbladder: A case report.

BACKGROUND: Neuroendocrine carcinoma is rare with a proportion of less than 2% in gallbladder malignancies, cases of gallbladder neuroendocrine cell carcinoma coexisting with adenocarcinoma are exceptionally rare, and the prognosis is dismal. METHODS: Herein, we presented an unusual case of poorly differentiated gallbladder neuroendocrine cell carcinoma coexisting with poorly differentiated adenocarcinoma who survived 20 months after the multimodal treatment (MT) of extended surgery and postoperative chemotherapy. RESULTS: Our result indicated that for advanced gallbladder neuroendocrine cell carcinoma coexisting with adenocarcinoma, MT including extended surgical approach combined with postoperative chemotherapy may contribute to a relatively good survival outcome. CONCLUSION: MT may contribute to a relatively good survival outcome for advanced gallbladder neuroendocrine cell carcinoma coexisting with gallbladder adenocarcinoma.

[Vertical Distribution and Pollution Risk Assessment of Nitrogen, Phosphorus, and Organic Matter in Sediment of Inflowing Rivers of Erhai Lake Estuarine Wetland in Wet and Dry Seasons].

The estuarine wetland of Luoshijiang River, which flows into Erhai Lake in Yunnan province, was regarded as the typical research object. Sediment samples in the estuarine wetland of Luoshijiang River were collected by the cylindrical sediment sampler and the samples were from surface layer (0-10cm), middle layer (10-30 cm), and bottom layer (30-60 cm). The vertical distribution of total nitrogen, total phosphorus, and total organic material in dry and rainy season was analyzed and contamination risk assessment of sediment was accomplished. The results showed that: ① In dry season, average concentrations of total nitrogen in sediment on surface, middle and bottom layer at estuarine wetland of Luoshijiang River were respectively 1.734, 1.453 and 1.255 g·kg-1, the average concentrations of total phosphorus were respectively 1.085, 1.034 and 0.992 g·kg-1, the average concentrations of organic material were respectively 59.051, 47.730 and 42.133 g·kg-1. In rainy season, average concentrations of total nitrogen in sediment on surface, middle and bottom layer at estuarine wetland of Luoshijiang River were respectively 1.147, 0.948 and 0.895 g·kg-1, the average concentrations of total phosphorus were respectively 0.599, 0.523 and 0.519 g·kg-1, the average concentrations of organic material were respectively 53.098, 46.897 and 43.395 g·kg-1. ② Single factor pollution index, organic nitrogen and organic index assessment were used to evaluate the contamination level of sediment, the assessment results showed that: In dry season, the contamination level of nitrogen and phosphorus in sediment was on severe level, organic material contamination was on medium level. In rainy season, the contamination levels of ON and OI were medium, and that of PI was mild. Generally, the contamination levels of nitrogen, phosphorus and organic material in sediment tendency were as follows: dry season>rainy season, and the pollution index of each layer followed the sequence of surface layer > middle layer > bottom layer, and the most extraordinary contamination of them on surface layer was uncovered whatever the season was. ③ Pollution origin of estuarine wetland of Luoshijiang River in dry and wet season had great differences, it was mainly the exogenous source in dry season, but it was the internal source in wet season; The potential risk of release of nutrient salt of sediment in dry season was higher than the wet, and the risk in the surface layer was the highest.

[Clinical analysis of the self made needle guide for the treatment of odontoid fracture].

OBJECTIVE: To analyze the curative effects and feasibility of the self regulating simple localizer through anterior approach for the treatment of odontoid fracture in adults. METHODS: From June 2010 and December 2012, 6 patients with odontoid fracture underwent an anterior operation using a single hollow screw located by the self regulating simple localizer. There were 4 males and 2 females, aged from 28 to 55 years old with an average of 39.1 years. The injuries were caused by traffic accidents in 4 cases and falling injury from high in 2 cases. According to the classification of Anderson, 4 cases were type II and 2 cases were simple type III. All the patients underwent operations in 5 to7 days after injury with the mean of 5.9 days. None of the patients had a spinal cord injury. The safety and feasibility of the self made localizer were observed in follow up for fracture healing and clinical effects. RESULTS: All the operations were successful with an average time of 50 min (ranged from 45 to 55 min) and the mean bleeding volume was 25 ml(ranged from 20 to 30 ml). No injuries of esophagus, trachea or nerve were found. All the patients were followed up from 8 to 16 months and all fractures were obtained bone healing. The flexion extension radiograph showed a well stability of atlantoaxial joint in last followed up. CONCLUSIONS: The self regulating simple localizer is a minimally invasive, short time and safe method in treating odontoid fractures through anterior operation with hollow screw. It may be a reliable choice while without a professional localizer.

The serum levels of MMP-9, MMP-2 and vWF in patients with low doses of urokinase peritoneal dialysis decreased uremia complicated with cerebral infarction.

To investigate the effect of MMP-9, MMP-2 and vWF in patients with low doses of urokinase peritoneal dialysis decreased uremia complicated with cerebral infarction. 112 cases of uremia complicated with cerebral infarction were randomly divided into the peritoneal dialysate with urokinase treatment group (66 cases) and the conventional treatment group (46 cases). At the same time, 50 cases of healthy people who were more than 45 years old were enrolled in the control group. The basic treatment in both treatment groups was the same. In urokinase therapy group based on the conventional treatment, urokinase was added into peritoneal dialysis fluid, and changes of serum MMP-9, MMP-2 and vWF were observed by drawing blood at different time points within 8 weeks. The changes of serum MMP-2, MMP-9 and vWF were detected by enzyme-linked immunosorbent assay. At the time of the onset of uremia complicated with cerebral infarction patients the serum MMP-9, MMP-2, vWF were significantly higher (P<0.05, P<0.05, P<0.01). Conventional antiplatelet therapy in brain protection only reduce MMP-9 to the normal range (P>0.05) within 8 weeks. But the MMP-2 and vWF cannot be reduced to the normal range (P<0.01, P<0.01). Low doses of urokinase can reduce MMP-9 (7 d) and MMP-2 (14 d) to the normal range (P>0.05, P>0.05) at the early stage and decrease the vWF to a normal range within 8 weeks (P>0.05). At the time of the onset of uremia complicated with cerebral infarction patients the serum MMP-9, MMP-2 and vWF increased significantly. Low doses of urokinase dialysis can reduce serum MMP-9, MMP-2, and vWF in acute uremia complicated with cerebral infarction without recurrence of cerebral infarction and cerebral hemorrhagic transformation, indicating that low dose of urokinase peritoneal dialysis may have a certain effect on the early treatment of this disease.

Identification and characterization of the antifungal substances of a novel Streptomyces cavourensis NA4.

A new actinomycete strain NA4 was isolated from a deep-sea sediment collected from the South China Sea and showed promising antifungal activities against soilborne fungal pathogens. It was identified as Streptomyces cavourensis by morphological, physiological, and phylogenetic analyses based on its 16S rRNA gene sequence. The main antifungal components were isolated and identified from the fermentation culture as bafilomycins B1 and C1. These compounds exhibited significant antifungal activities and a broad antifungal spectrum. The results suggest that the Streptomyces cavourensis NA4 and bafilomycins B1 and C1 could be used as potential biocontrol agents for soilborne fungal diseases of plants.

New spirotetronate antibiotics, lobophorins H and I, from a South China Sea-derived Streptomyces sp. 12A35.

Strain 12A35 was isolated from a deep-sea sediment collected from the South China Sea and showed promising antibacterial activities. It was identified as Streptomyces sp. by the 16S rDNA sequence analysis. Bioassay-guided fractionation using HP20 adsorption, flash chromatography over silica gel and octadecylsilyl (ODS) and semi-preparative HPLC, led to the isolation and purification of five metabolites from the fermentation culture of 12A35. Two new spirotetronate antibiotics, lobophorins H (1) and I (2), along with three known analogues, O-ß-kijanosyl-(1→17)-kijanolide (3), lobophorins B (4) and F (5) were characterized by 1D, 2D-NMR and MS data. These compounds exhibited significant inhibitory activities against Bacillus subtilis. Compounds 1 and 5 exhibited moderate activities against Staphylococcus aureus. In particular, the new compound lobophorin H (1) showed similar antibacterial activities against B. subtilis CMCC63501 to ampicillin.

Potent toxic macrocyclic trichothecenes from the marine-derived fungus Myrothecium verrucaria Hmp-F73.

Activity-guided fractionation of a methanol extract from the culture broth of Myrothecium verrucaria Hmp-F73, a fungus associated with the sponge Hymeniacidon perleve, afforded six macrocyclic trichothecenes, verrucarin J (1), 8-hydroxyverrucarin J (2), verrucarin A (3), 8-acetoxyroridin H (4), isororidin E (5), and roridin E (6), along with trichoverrin B (7). All seven metabolites displayed potent toxicity to the brine shrimp (Artemia salina). In addition, compounds 2, 3, and 6 showed weak phytotoxic activities against lettuce seeds. A preliminary structure-activity relationship of the metabolites is also discussed.

[Analysis of therapeutic effects of external fixator for the treatment of comminuted fracture of distal radius].

OBJECTIVE: To evaluate the clinical effects of external fixator in treating comminuted fracture of distal radius. METHODS: From Mar.2008 to Dec.2009, 37 patients with comminuted fracture of distal radius were treated with external fixator or assisted with Kirschner wire and T-shape locking compression plate (T-LCP) fixation. There were 14 males and 23 females, ranging in age from 30 to 79 years, with an average of 59.1 years. According to AO typing, type C1 was in 3 cases, type C2 was in 11 cases and type C3 was in 23 cases. Function of wrist joint and X-ray films were observed according to Gratland-Werley system at different months. RESULTS: All patients were followed up from 8 to 24 months with an average of 12 months. All fractures had healing with an average time of 8 weeks. According to Gratland-Werley system, 16 cases obtained excellent result, 17 good, 4 fair, the rate of excellent and good was 89.0%. CONCLUSION: External fixator can obtained satisfactory clinical effect in treating comminuted fracture of distal radius.