Performance of Arma chinensis reared on an artificial diet formulated using transcriptomic methods.

An artificial diet formulated for continuous rearing of the predator Arma chinensis was inferior to natural prey when evaluated using life history parameters. A transcriptome analysis identified differentially expressed genes in diet-fed and prey-fed A. chinensis that were suggestive of molecular mechanisms underlying the nutritive impact of the artificial diet. Changes in the diet formulation were made based on the transcriptome analysis and tested using life history parameters. The quantity of pig liver, chicken egg, tuna fish, biotin, nicotinamide, vitamin B6, thiamine, riboflavin, vitamin C, L-glutamine, and sucrose was reduced, and wheat germ oil, calcium pantothenate and folic acid were increased. Ecuadorian shrimp was added as a partial substitute for tuna fish. Several parameters improved over six generations, including increased egg viability, and decreased egg and adult cannibalism. Additionally, several parameters declined, including longer developmental times for 2nd-5th instars, and decreased nymphal weights. The improvements in life history parameters support the use of transcriptome analyses to help direct formulation improvements. However, the decline in some parameters suggests that additional information, e.g., proteomic data, may be useful as well to maximize diet formulations.

LncRNA H19 serves as a ceRNA and participates in non-small cell lung cancer development by regulating microRNA-107.

OBJECTIVE: To investigate whether lncRNA H19 can regulate NF1 expression through competitive binding to microRNA-107, thereby participating in the occurrence and development of non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Expression levels of H19 and NF1 in NSCLC tissues, paracancerous tissues and NSCLC cell lines were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). The binding condition of microRNA-107, H19 and NF1 was detected by dual-luciferase reporter gene assay. Corresponding lentiviruses of H19 were constructed. The regulatory effects of H19 on proliferative and migratory abilities of A549 cells were detected by cell counting kit-8 (CCK-8) and transwell assay, respectively. Rescue experiments were conducted to explore the regulatory interaction between H19 and microRNA-107 in A549 cells. RESULTS: H19 and NF1 were highly expressed in NSCLC tissues and NSCLC cell lines (A549 and HCC823) than those of controls. Overexpressed H19 increased proliferative and migratory abilities of A549 cells. Dual-luciferase reporter gene assay demonstrated that H19 regulates NF1 expression through competitive binding to microRNA-107, thereafter participating in NSCLC development. CONCLUSIONS: H19 is highly expressed in NSCLC, which promotes NSCLC development by regulating NF1 via competitive binding to microRNA-107.

[Relationship between coronary tortuosity and coronary microvascular disease].

Objective: To explore the relationship between coronary tortuosity and coronary microvascular disease (CMVD). Methods: Patients with typical angina symptoms and without serious coronary artery stenosis by coronary angiography were enrolled from June 2014 to December 2016, and CMVD was diagnosed by single photon emission tomography (SPECT). According to the SPECT results, patients were divided to the CMVD group and non-CMVD group. The baseline clinical characteristics, results of coronary angiography were compared between the two groups. The logistic analysis was used to analyze the relationship between coronary tortuosity and CMVD. Result: A total of 117 cases were enrolled, with 69 cases in the CMVD group and 48 cases in the non-CMVD group. No differences were found in gender distribution, age, hypertension, lipid abnormality, hyperuricemia and uses of statins between the two groups (all P>0.05). Incidence of diabetes (78.26%(54/69) vs. 35.42% (17/48) , P<0.05), hs-CRP ((4.29±2.15)mmol/L vs. (2.63±1.20)mmol/L, P<0.001), LDL-C ((2.98±0.96)mmol/L vs. (2.52±0.83)mmol/L, P=0.008) and homocysteine ((13.7±5.61)mmol/L vs. (11.5±4.38)mmol/L, P=0.025) levels were higher in the CMVD group than in the non-CMVD group. The data derived from echocardiographic examination were similar between the two groups. The Corrected TIMI frame counts were higher in the CMVD group than in non-CMVD group (LAD: 31.56±4.92 vs. 27.31±3.75, LCX: 29.47±4.18 vs. 26.62±3.19, RCA: 29.09±5.05 vs. 26.24±3.28, all P<0.001). The incidences of coronary atherosclerosis (76.81% (53/69) vs. 27.08% (13/48) , P<0.001) and coronary tortuosity ( (60.87% (42/69) vs. 33.33% (16/48) , P=0.035) were also higher in the CMVD group than in non-CMVD group. Logistic analysis found that coronary tortuosity (OR=6.111, 95%CI 2.707-13.794, P<0.001), diabetes (OR=6.565, 95%CI 2.883-14.948, P<0.001) and coronary atherosclerosis (OR=8.918, 95%CI 3.822-20.808, P<0.001) were independent risk factors of CMVD. Conclusion: Coronary tortuosity, diabetes and coronary atherosclerosis are related to CMVD in this patient cohort.

Performance and Cost Comparisons for Continuous Rearing of Arma chinensis (Hemiptera: Pentatomidae: Asopinae) on a Zoophytogenous Artificial Diet and a Secondary Prey.

The impact of a zoophytogenous, insect-free artificial diet and a secondary prey, pupae of Chinese oak silk moth Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), on the developmental rate, life history parameters, and fertility was examined for F6, F9, and F12 consecutive generations for domesticated Arma chinensis (Fallou) (Heteroptera: Pentatomidae). This study showed that when fed an insect-free artificial diet during both the nymphal and adult stages, developmental times were prolonged, and fecundity, egg viability, net reproductive rates (R0), and intrinsic rates of increase (rm) declined. As a result, the cost to rear A. chinensis on the artificial diet approached 1.7 times the cost of rearing A. chinensis on pupae of A. pernyi. Future diet improvements should attempt to reduce developmental time, increase fecundity, and egg viability and use less costly nutrient sources.

Resveratrol, an activator of SIRT1, upregulates AMPK and improves cardiac function in heart failure.

Reduced AMP-activated protein kinase (AMPK) expression has been shown to play a significant role in the cardiac dysfunction in heart failure. This study was designed to examine the effect of resveratrol, a potent activator of silent information regulator (SIRT1), on cardiac function and AMPK expression in heart failure. Adult male rat left anterior descending arteries were ligated, and they were fed with either a regular diet or a diet enriched with resveratrol. Heart failure was produced by myocardial infarction, and was associated with markedly increased AMPK and SIRT1 protein levels. Resveratrol treatment had a tremendous beneficial effect, both in terms of improving AMPK expression and on cardiac function. Decreased cardiac function and AMPK expression were also found in SIRT1 knockout (+/-) mice. In cultured cardiomyocytes, resveratrol increased AMPK and SIRT1 expressions, and overexpression of SIRT1 was found to be sufficient to activate AMPK in H9c2 cells. In contrast, pretreatment of cardiomyocytes with an SIRT1 antagonist, nicotinamide, blocked these beneficial effects of resveratrol. Therefore, the protective effects of resveratrol were found to be dependent on its ability to activate SIRT1 and improve AMPK expression. These results demonstrated that in heart failure, the enzymatic activity of cardiac SIRT1 is increased, which contributes to increased expression of AMPK, and resveratrol enhances the expression of AMPK and improves cardiac function through the activation of SIRT1.

Protective effects of MCI-186 on transplantation of bone marrow stromal cells in rat ischemic stroke model.

Transplantation of bone marrow stromal cells (BMSCs) is a potential therapy for ischemic stroke, but poor environmental conditions in brain lesions, such as insufficient nutrition and oxygen free radical toxicity, limit the efficacy of stem cell therapy. Here, we hypothesized that MCI-186, a free radical scavenger, would have protective effects on transplantation of BMSCs in a rat ischemia model. In vitro, flow cytometry showed the apoptotic rates of BMSCs after simulated ischemia-reperfusion (I/R) injury was significantly decreased when treated with MCI-186 (P<0.01). In vivo, rat transient middle cerebral artery occlusion (MCAO) model was established. Two separate MCAO groups were administered with either MCI-186 or phosphate-buffered solution (PBS) immediately after artery occlusion. MCI-186 significantly up-regulated the secretion of brain-derived neurotrophic factor, vascular endothelial growth factor and superoxide dismutase in ischemic brain, while malondialdehyde decreased and neuronal apoptosis was inhibited. Furthermore, another four MCAO groups were administered with either PBS, MCI-186, BMSCs (2×10(6)) or a combination of MCI-186 and BMSCs. When compared with BMSCs or MCI-186 monotherapy, combination therapy significantly improved functional restoration, decreased infarct volume, and increased the number of engrafted-BMSCs and neurons in ischemic brain. The number of engrafted-BMSCs and neurons was significantly correlated with functional outcomes. This study suggests that MCI-186 may improve the environment of the injured brain, enhance the survival of engrafted-BMSCs and neurotization in ischemic brain and produce protective effects on BMSCs transplantation.

The noggin2 gene of Gekko japonicus (Gekkonidae) is down-regulated in the spinal cord after tail amputation.

The cDNA encoding noggin2 protein was obtained from the brain and spinal cord cDNA library of Gekko japonicus. The size of the noggin2 transcript and its expression in different tissues were analyzed by Northern blot analysis. In situ hybridization revealed positive hybridization signals in both gray and white matter of the spinal cord. Changes in noggin2 expression in the spinal cord after tail amputation were examined by real-time PCR. The noggin2 was expressed in the normal spinal cord and down-regulated three days after tail amputation, reaching the lowest level at two weeks, during the time course when we followed the expression levels. We concluded that the expression of noggin2 is affected by the process of spinal cord injury and regeneration.

The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo.

AIM: In the present study, we investigated the biodegradation of the fibers of chitosan and its acetylated derivatives in vitro and in vivo. METHODS: A series of chitosan fibers, with acetylation degrees of 7.7%, 21.6%, 40.9%, 61.2%, 82.5% and 93.4%, were obtained by acetylating chitosan filament with acetic anhydride, and were investigated by FT-IR analysis, elemental analysis and scanning electron microscopy analysis. RESULTS: The in vitro experimental data indicated that the degradation rate of chitosan fiber was strongly dependent on the degree of acetylation, and the degradation rate increased with an enhancement of the acetylation degree of chitosan fibers. In vivo degradation experiment evaluated by light microscopy as well as scanning electron microscopy, was studied by implanting the fibers between the two nerve stumps of the rat sciatic nerve gap (6 months). The findings demonstrated that acetylation degree could influence the degradation rate of chitosan fibers in vivo. CONCLUSION: These results suggested that acetylated chitosan (chitin) fibers were more biodegradable than chitosan and the biodegradation rate of chitin fiber can be controlled to desirable extent by the variation of acetylation degree.

Disruption of granules by hydrodynamic force in internal circulation anaerobic reactor.

A process of granule disruption by hydrodynamic force is discussed in this paper. Shear force and attrition among granules originated from hydrodynamic force are the main causes of the disruption. Since it is positively correlated to the attrition force, the shear force is utilized to describe the effect of hydrodynamic force on granule disruption. In the experiment, when increase rate of average shear rate (IRgamma) of the 1st stage is about 0.2 s(-1) x d(-1), the granules are disrupted; while re-granulation could develop when IRgamma is about 0.07 s(-1) x d(-1); even when the shear rate is as high as about 30s(-1), the granulation rate keeps stably at a relatively high level, which shows that granules could bear the high hydrodynamic force only if it increases by low increase rate. The experimental results would be valuable for the operation and controlling of the upflow reactors.

Determination of ketorolac enantiomers in plasma using enantioselective liquid chromatography on an alpha 1-acid glycoprotein chiral stationary phase and ultraviolet detection.

A chirally selective high-performance liquid chromatographic assay was developed to measure the R(+) and S(-) enantiomers of ketorolac in plasma for pharmacokinetic studies. Naproxen sodium [S(+) enantiomer] (10 micrograms) was used as an internal standard. Plasma samples (0.5 ml) were acidified (50 microliters of 4 M H3PO4 to pH 1.5), extracted into 0.4 ml of 10% pentan-2-ol in hexane and back-extracted into 0.15 ml of base (20 mM NaOH pH to 7-8), of which samples (5 microliters) were chromatographed on a 100 x 4 mm I.D. column packed with an HPLC chiral stationary phase based upon immobilized alpha 1-acid glycoprotein (Chiral AGP-CSP) with 4% propan-2-ol in 0.1 M NaH2PO4 pH 5.5, at 0.9 ml/min. Detection was at 325 nm and run time was 10 min. Retention times of R- and S-ketorolac and of S(+)-naproxen were 3.3, 4.8 and 6.4 min, respectively. The metabolite p-hydroxyketorolac was not resolved enantiomerically and had a retention time of 2.2 min. The assay was linear over the range 0.5-10 mg/l, with precisions < 5% C.V. Good separations (alpha > 1.35) and resolutions (Rs > 3.23) between peaks were achieved. The sensitivity could be extended to 35 micrograms/l with less precision by increasing the injection volume to 100 microliters.

Rapid immunostaining of live nerve for identification of sensory and motor fasciculi.

Six New Zealand white rabbits were anesthetized with pentobarbital, and sciatic nerves were exposed and cut at the mid thigh. Both proximal and distal ends were incubated directly with Blue-SAb in a micropipe of 5 mm diameter covering the nerve trunk ends for 30 minutes at room temperature. After removal of the micropipe, the nerve ends were washed with physiological saline. Blue-stained fasciculi, i.e., sensory fasciculi were seen among unstained ones under the operating microscope. This method requires no histological sections. Neural cells of the spinal cord and ganglion were cultured in RPM11640 medium containing Bright Blue. The growth and metabolism of the neural cells were tested by MTT assay and their morphology was observed. Statistical difference between the experiment and control groups was determined, indicating that Bright Blue had no effect on the neural cells and their repairing power. This rapid immunostaining technique offers a good approach for the identification and accurate coaptation of sensory fasciculi in peripheral nerve repair.

Isolation of a strain of Hantaan virus from peritoneal exudate cells of a patient with hemorrhagic fever with renal syndrome.

A new strain of Hantaan virus (HTNV), GH716, was isolated from the peritoneal exudate cells (PEC) of a patient with severe hemorrhagic fever with renal syndrome (HFRS). The isolate was propagated in Vero E6 cells. At each passage the virus-infected cells were examined for HTNV with immunofluorescence technique using monoclonal antibodies (McAb) 25-1 and 84-1 against HTNV. From passage 5 on, fluorescent intensity tended to stabilize at (+++) and infectivity titer reached 10(5) TCID50/ml. Using 14 McAb to 76-118 and BI strains of HTNV, we found that strain GH716 was antigenically similar to strain 76-118 (Apodemus type) and different from strain R22 (Rattus type), suggesting that GH716 may fall into the Apodemus type of HTNV. This is the first isolation of an HTNV from human PEC collected on the tenth day of illness. The successful isolation of strain GH716 may provide an alternative source of obtaining HTNV during the later stages of HFRS.

Hemorrhagic fever with renal syndrome. Separation of human peripheral blood T and B cells and detection of viral antigen.

In this study, total blood lymphocytes were prepared from patients with hemorrhagic fever with renal syndrome (HFRS) by a density gradient on Ficoll-Hypaque. T and B cells were then purified by passing the total lymphocytes over a nylon wool column. The purities of total lymphocytes, T cells and B cells were 97.8 +/- 2.3%, 91.6 +/- 4.5% and 74.2 +/- 12.1%, respectively. Also, after modification of cell fixation and smear drying, the number of cells were increased and the time needed for slide preparation was shortened. Detection of viral antigen by immunofluorescence assay using monoclonal antibodies to Hantavirus (HTNV) showed that the total lymphocytes. T cells and B cells were infected by HTNV during the early stages of HFRS and no specific fluorescence was seen in the cells from the late diuretic phase to convalescent phase. The results suggest that virus replication in blood lymphocytes may partly contribute in the early stages to the impairment of cell immune response and in vivo spread of HTNV to its target sites.

[Effects of stopping water fluoridation on prevalence of dental caries in children].

Fluoridation of public water supply in Fangcun Guangzhou was carried out from 1965 to 1983. Data of dental caries of children aged 3-6 in Fangcun and other statistical data were in May 1986 and January 1988, which were 31 months and 52 months respectively after stopping water fluoridation in October 1983. Compared with that during the water fluoridation, the result of survey in 1986 showed that prevalence of dental caries and other statistical figures increased significantly (P less than 0.01) in the 3-year-old group, but non-significantly in the groups of children aged 4-6 because they were born during the water fluoridation period. The result of survey in 1988 demonstrated significant increase of dental caries in the groups of children aged 3-5, particularly the interdental caries of front teeth which went up most noticeably, but non-significantly in the 6-year-old groups for the same reason. The results confirmed that systemic effects of fluoridation were more important and the prevention of interdental caries was best in water fluoridation programs.

Hemorrhagic fever with renal syndrome: separation of human peripheral blood T and B cells and detection of viral antigen.

Total lymphocytes in peripheral blood were prepared from patients with hemorrhagic fever with renal syndrome by a density gradient on Ficoll-Hypaque, and then T and B cells purified by passing the lymphocytes over a nylon column with modifications. Also, after modification of cell fixation and use of a spinner for smear drying, the number of cells were increased and the duration for slide preparation was shortened, thus resulting in quality slides. Detection of viral antigen by immunofluorescence assay using monoclonal antibodies to Hantavirus (HTNV) showed that the total lymphocytes, T cells and B cells were infected with HTNV during the early stages of the illness and no specific fluorescence was seen in the cells from the late diuretic phase to convalescent phase.