Expression of chicken epidermal growth factor (cEGF) in Escherichia coli regulates the microflora structure of the duodenum to improve growth performance and intestinal morphogenesis in broilers.

1. A study used gene synthesis to obtain the functional domains of chicken epidermal growth factor (cEGF) and examined their impact on broiler growth performance, small intestinal morphology, digestive enzyme activities in the intestinal contents and the structure of duodenal microflora.2. The pET-32a-cEGF recombinant expression vector was constructed. The specific band at 26 KDa was shown by SDS-PAGE analysis and WB results. The purified protein content was shown to be 1687 µg/ml by assay.3. A total of 180 healthy, one-day-old Arbor Acres male, white-feathered broilers were randomly divided into three dietary treatment groups (six replicate pens, 10 birds per replicate): A control diet (ND); cEGF diet (cEGF), control supplemented with 250 mg/kg cEGF and the control diet (CD) supplemented with 250 mg/kg chlortetracycline.4. The results showed that feeding the cEGF and CD diet reduced FCR of broilers aged 1-21 d, average daily feed intake (ADFI) at 22-42 d, and the FCR in the whole period (1-42 d; p < 0.05). Compared with the ND group, the cEGF diet increased duodenal α-amylase and alkaline phosphatase activities in the 1-21 d, duodenal lipase, alkaline phosphatase, and ileal alkaline phosphatase activities in the post-period and increased villus height in the duodenum and ileum (p < 0.05). In addition, the ACE and Chao1 index for the birds fed cEGF were higher than the ND group (p < 0.05). At the phyla level, Firmicutes and Proteobacteria were dominant in all groups. At the genus level, the dominant genus was Lactobacillus. The LEfSe analysis showed that the cEGF group was enriched by 11 species including Brevibacillus, Eisenbergiella, Cloacibacterium, Butyricoccus spp.5. The addition of 250 mg/kg cEGF to the diet can increase growth performance by improving intestinal development and digestive enzyme activity, which may be related to the duodenal intestinal microflora. Therefore, cEGF is an effective alternative to antibiotics in broiler farming.

MiRNA-155 promotes the invasion of colorectal cancer SW-480 cells through regulating the Wnt/ß-catenin.

OBJECTIVE: To investigate the role of microRNA-155 (miR-155) in the potential invasion of colon cancer cell and the underlying mechanism. PATIENTS AND METHODS: The expression level of miR-155 in colon cancer and adjacent normal tissues was detected by Real-time quantitative polymerase chain reaction (RT-PCR). miR-155 mimics (miR-155), or siRNA against ß-catenin (ß-catenin siRNA), was transfected into human colon cancer cell line SW-480 using Lipofectamine 2000, respectively. RT-PCR was used to measure the expression levels of miR-155 and ß-catenin mRNA, and ß-catenin protein expression level was detected by Western blot. The in-vitro cell invasion abilities were determined by transwell invasion assays after up-regulating miR-155 or knocking down of ß-catenin. RESULTS: MiR-155 directly regulates ß-catenin at the transcriptional level, and promotes the invasion potential of colon cancer cell, at least partly through the upregulation of ß-catenin. CONCLUSIONS: The findings of this study suggest that miR-155 and ß-catenin may have a unique potential as a novel biomarker candidate for diagnosis and treatment of tumor metastasis.

[Silencing of vacuolar ATPase c subunit ATP6V0C inhibits invasion of prostate cancer cells].

OBJECTIVE: Vacuolar ATPase (V-ATPase) was found within the membranes and internal organelles of a vast array of eukaryotic cells, and was related to various kinds of highly metastatic tumors. LASS2/TMSG1 gene was a novel tumor metastasis suppressor gene cloned from human prostate cancer cell line PC-3M in 1999 by our laboratory. It was found out that protein encoded by LASS2/TMSG1 could interact with the c subunit of V-ATPase (ATP6V0C). In this study, To use RNA interference to suppress the expression of ATP6V0C and try to further investigate the molecular mechanism of ATP6V0C in tumor metastasis and its relationship with LASS2/TMSG1 gene. METHODS AND RESULTS: The expression level of ATP6V0C mRNA and protein in high metastatic potential prostate cancer cell lines (PC-3M-1E8 and PC-3M) was significantly higher than that in low metastatic potential prostate cancer cell lines (PC-3M-2B4 and PC-3), the expression level in PC-3M-1E8 being the highest. Follow-up tests selected PC-3M-1E8 cells for gene silencing. The expression and secretion of MMP-2 and the expression of MMP-9 in ATP6V0C siRNA transfected PC-3M-1E8 cells displayed no obvious change, but the activity of secreted MMP-9 was abated noticeably compared with the controls (P<0.05). Extracellular hydrogen ion concentration and V-ATPase activity in interference group were both reduced significantly compared with the controls (P<0.05). The migration and invasion capacity of ATP6V0C siRNA interfered cells in vitro were diminished significantly compared with the controls (P<0.05). Furthermore, a dramatic reduction of LASS2/TMSG1 mRNA and protein level after transfection of siRNA in PC-3M-1E8 cells was discovered (P<0.05). Confocal immunofluorescence showed a vast co-localization of ATP6V0C protein and LASS2/TMSG1 protein in plasma and membrane. The co-localization signals of control group were much stronger than those of interference group. CONCLUSION: Specific siRNA silencing of ATP6V0C gene inhi-bits the invasion of human prostate cancer cells in vitro by mechanism of inhibiting V-ATPase activity and then reducing the extracellular hydrogen ion concentration, inhibiting MMP-9 activation and affecting ECM degradation and reconstruction. Meanwhile, ATP6V0C and LASS2/TMSG1 have interaction and it is likely that ATP6V0C functions as a feedback regulator of LASS2/TMSG1.

[Analysis on occupational noise-induced hearing loss of different type workers in underground mining].

Objective: To investigate hearing loss status of blasters, drillers mechanics and so on in underground mining, and put forward suggestion diagnosis of occupational explosive deafness and occupational deafness. Methods: Underground excavation workers in a metal mine were recruited in this study, those with a history of ear disease and non-occupational deafness were all excluded. Finally, the features of pure tone audiometry of 459 noise-exposed workers were analyzed. Results: High-frequency hearing loss occurred on 351workers and the positive detection rate was 74.29%, workers who had both high-frequency and linguistic frequency hearing loss were 51 and the positive detection rate was 11.11%. The positive detection of high-frequency hearing loss in right ear (χ(2)=9.427 and P= 0.024) and in left ear (χ(2)=14.375, P=0.002) was significantly different between different exposure age groups. The positive detection of high-frequency hearing loss of driving group was the highest, followed by blasting group, mining group and machine repair group. The characteristics of the hearing loss caused by drilling noise of the blasting workers with no accident occurred were in line with that of noise-induced hearing loss. Conclusion: The diagnosis grading should be carried out according to the diagnostic criteria of occupational noise-induced deafness for the employees who engaged in the blasting operation with no record of blast accident.

Correlation between the NPPB gene promoter c.-1298 G/T polymorphism site and pulse pressure in the Chinese Han population.

The aim of this study was to investigate the correlation between the natriuretic peptide precursor B (NPPB) gene single nucleotide polymorphism (SNP) c.-1298 G/T and pulse pressure (PP) of the Chinese Han population and the association between genotype and clinical indicators of hypertension. Peripheral blood was collected from 180 unrelated patients with hypertension and 540 healthy volunteers (control group), and DNA was extracted to amplify the 5'-flanking region and 2 exons of the NPPB gene by polymerase chain reaction; the fragment was sequenced after purification. The clinical data of all subjects were recorded, the distribution of the NPPB gene c.-1298 G/T polymorphism was determined, and differences in clinical indicators between the two groups were evaluated. The mean arterial pressure PP, and creatinine levels were significantly higher in the hypertension group than in the control group (P<0.05), but no other clinical indicators differed between the groups. There were no significant differences in genotype frequency and distribution of the NPPB gene c.-1298 G/T polymorphism between the hypertension group and the control group (P>0.05); in the control group, the mean PP of individuals with the SNP c.-1298 GG genotype was greater than that of individuals with the GT+TT genotype (P<0.05). In conclusion, there was no significant correlation between the NPPB gene c.-1298 G/T polymorphism and the incidence of essential hypertension in the Han population; however, the PP of the SNP c.-1298 GG genotype was greater than that of the GT+TT genotype in the control group.

Note: phase retrieval method for analyzing single-phase displacement interferometry data.

We present a phase retrieval method (PRM) for analyzing single-phase displacement interferometry measurements on rapidly changing velocity histories, including photon Doppler velocimetry (PDV). PRM identifies the peaks and valleys as well as zero-crossing points in a PDV time series, performs normalization and extracts point-by-point phase and thus velocity information. PRM does not require a wide time window as in sliding window Fourier transformation, and thus improves the effective temporal resolution. This method is implemented in analyzing PDV data obtained from gas gun experiments, and validated against simultaneous measurements with velocity interferometer system for any reflector.

Identification of a novel frame-shift mutation in PRSS1 gene in Han patients with autoimmune pancreatitis.

OBJECTIVE: To detect mutations of trypsinogen gene (PRSS1) in patients with autoimmune pancreatitis (AIP) and to determine the underlying pathogenesis. METHODS: DNA sequencing was used to detect full-length of PRSS1, cystic fibrosis transmembrane conductance regulator (CFTR), and pancreatic secretory trypsin inhibitor (SPINK1) genes mutations in an AIP family and a sporadic case and 520 normal controls. Furthermore, a mutant-expressing system was constructed for functional confirmation. RESULTS: For the first time, we report a deletion mutation at exon 2 of PRSS1 gene (IVS 2 +56_60 del CCCAG) which encoded a truncated PRSS1 protein without trypsinogen activation peptide (TAP). Vitro functional study suggested the identified mutation would result in loss of PRSS1 activity. Mutant trypsinogen activated at a faster rate than wild-type trypsinogen in the autoactivation experiment. Histopathologic examination revealed the ratio of IgG4/IgG-positive plasma cells exceeded 0.455 in pancreas, and the patients responded to glucocorticoids. CONCLUSION: PRSS1: IVS 2 +56_60 del CCCAG is a noval mutant which may contribute to AIP pathogenesis.

Multi-walled carbon nanotube-supported metal-doped ZnO nanoparticles and their photocatalytic property.

A simple and versatile approach has been developed to synthesize multi-walled carbon nanotubes/metal-doped ZnO nanohybrid materials (MWNT/M-doped ZnO) by means of the co-deposition method. The experimental results illuminate that MWNTs can be modified by metal-doped ZnO nanoparticles at 450 °C, such as Mn, Mg, and Co elements. Furthermore, the MWNT/Mg-doped ZnO hybrids have been proven to have a high photocatalytic ability for methyl orange (MO), in which the degraded rate for MO reaches 100 % in 60 min. The enhancement in photocatalytic activity is attributed to the excellent electriconal property of MWNTs and Mg-doping. The resultant MWNT/Mg-doped ZnO nanohybrids have potential applications in photocatalysis and environmental protection.

Chemical and mineralogical composition of the Mongolian rural soils and their uranium sorption behavior.

Distribution of uranium (VI) between soil solids and solutions is a key parameter in assessing the risk to the biosphere of disposing uranium-rich waste products from nuclear plants as well as uranium (U) ore mining. Both of these topics have recently been brought to public attention in Mongolia. Regional background levels of soil elements are an important dataset for accessing the actual environmental situation and monitoring pollution levels. Little information, however, is available on background concentrations of various elements in Mongolian soils. Thirteen rural soils were sampled from six provinces in Mongolia, and the concentrations of macro-, micro- and trace elements were measured. The values obtained served as a reference (baseline) for uncontaminated soils. The soils were characterized with slightly acidic to strongly alkaline pH values. With the exception of the sample from a western province, all the soils investigated contained little organic matter. The content of soil elements did not vary widely among geographical regions. The concentration of most micro elements was within the range of worldwide soil values but the value for Zn tended to be moderately higher. The U (VI) sorption into the soils was investigated using the batch technique and the (237)U radionuclide tracer, produced by the photo fission reaction (238)U(γ, n) (237)U at an electron accelerator. The (237)U distribution coefficient (K(d)), derived from the sorption isotherms, was related to solution pH and varying from 9 to 2547 mL g(-1) when the pH ranged between 3 and 7.7. The sorption process was interpreted in terms of the formation of different U (VI) species at given concentrations, calculated using the Speciation program with and without carbonate in the system. The U sorption isotherm displayed two general patterns: one where sorption decreased as solution pH increased, showing a maximum at pH 3, and another pattern revealed an adsorption maximum at pH 5 and then decreased up to pH 7.7 (the final solution pH). The observed decrease in K(d) when solution pH increased from 6 to 8 was consistent with the increased formation of soluble UO(2)(OH)(2) species. A linear negative correlation between lgK(d) and the solution pH was observed similarly to that reported for the soils with a pH ≥ 6.

Impact of a novel mutation in the 5'-flanking region of natriuretic peptide precursor B gene on the antihypertensive effects of sodium nitroprusside in patients with hypertension.

Our aim was to identify the possible mutations of the natriuretic peptide precursor B (NPPB) gene in a family with hereditary hypertension, and determine whether the mutations are associated with the antihypertensive effect of sodium nitroprusside. The subjects included one family with hereditary hypertension, 36 cases of sporadic hypertension and 120 healthy controls. The 5'-flanking sequence of NPPB was amplified with PCR, and the presence of mutations was analyzed by direct sequencing. Patients with hypertension were treated with sodium nitroprusside and blood pressure data and serum B-type natriuretic peptide (BNP) levels were measured. A novel complex mutation in 5'-flanking sequence of the NPPB gene was detected in three patients (II 2, III 2, and III 5) of the hypertension family, which included c.-1195_ -1176 insert 5'-CCTTCTTTCTTTCTTTCTTT-3', c.-1208 T>A, c.-1214 T>C, and c.-1216 T>A. Patients with this mutation were less sensitive to sodium nitroprusside treatment. Sporadic hypertension patients (without NPPB gene mutation) and patients with the c.-1181 T>A point mutation were sensitive to sodium nitroprusside treatment. BNP levels of patients with the complex mutation were significantly lower than that of sporadic hypertension patients and c.-1181 T>A mutation patients before and during the early stage of sodium nitroprusside treatment. The complex mutation of the NPPB gene might be an etiological factor of hereditary malignant hypertension, and it is associated with low sensitivity to the antihypertensive effect of sodium nitroprusside.

Distribution of heavy metals from iron bath-melting separation process applied to municipal solid waste incineration fly ash.

Fly ash generated from municipal solid waste (MSW) incineration is frequently classified as hazardous material and requires special disposal. The management of the large amount of fly ash has caused increasing problems in China. This work describes a novel approach for melting MSW incineration fly ash, and the distribution of heavy metals was characterized during the iron bath-melting separation process. Four hundred grams of pelletized fly ash was fed into the furnace in molten iron bath atmosphere. After the melting separation process, the distribution of heavy metals in samples and the leaching characteristics of the slag were investigated. The results indicated that iron bath-melting promoted the transfer of Cr, Mn and Cu from the slag phase to the iron phase, which also improved Zn and Pb volatilization. The leaching concentrations, determined by the Chinese Standard Method (rollover leaching procedure) of the target metals of the slag from leaching tests were lower than the Chinese regulatory thresholds. Therefore, this method was proposed as an environmentally friendly technology to achieve a satisfactory solution for MSW incineration fly ash management.

[Clinical study of TCM-WM on aplastic anemia complicated with hepatitis C].

The testing kit of second generation for serum anti-HCV was used in 82 cases of aplastic anemia (AA). The results showed that positive rate was 69.4% (43/62) in the patients of AA with transfusion, this was significantly higher than that in the patients of AA without transfusion. There was no difference of anti-HCV antibody positive rate between chronic AA and acute AA (P > 0.05), incidence rate of post-transfusion hepatitis C (PTHC) in AA was 33.9% (21/62), among which the incidence rate in acute and chronic AA were 68.8% (11/16) and 21.7% (10/46) respectively (P < 0.01). The anti-HCV positive patients were divided into two groups: PTHC and non-PTHC, there was no statistical difference of their transfusion volume, hemoglobin, white blood cell between these groups. Response rate of AA was lower in anti-HCV positive patients than that in negative patients (P < 0.05). Acute, icteric PTHC was predominant in patients with AA. The patients with AA complicated with PTHC was liable to bleed and be infected. PTHC has been an important complication in patients with AA. The better response was obtained by TCM-WM therapy in the patients.

Distribution and location of immunoreactive atrial natriuretic peptides in cochlear stria vascularis of guinea pig.

The distribution and location of atrial natriuretic peptides (ANP) in the cochlear stria vascularis of normal guinea pigs were studied to find out regional regulating factors of cochlear blood flow (CBF) and productive foundation of cochlear endolymph by ABC immunocytochemistry and immunoelectronmicroscopy. The ANP-IR products were seen in the stria vascularis of the 1-4 turns of the lateral wall of the cochlea, more markedly along its margin; and were negative in the spiral ligaments. Immunoelectron-microscopically, a plenty of sphericals of 250-350nm in diameter were noted in the cytoplasm of marginal and intermediate cells, but few in the basal cells. The results suggest that the cochlear stria vascularis had the function of secreting ANP in the guinea pig.

Duration of immunity following immunization with live measles vaccine: 15 years of observation in Zhejiang Province, China.

The duration of immunity following measles vaccination of 2882 immunized children has been investigated in a closed region of China for 15 years. A total of 1002 of the children were treated as primary immunization subjects, and 1547 as reimmunization subjects. These two cohorts were not in contact with known wild measles virus over the whole observation period, and the results obtained probably reflected the antibody responses to measles vaccine alone. The remaining 333 vaccinees came into contact with wild measles virus, and this permitted evaluation of the protective effect of the measles vaccines tested: 4 children experienced very mild clinical measles, and 329 experienced subclinical infection, including 12 who had had undetectable haemagglutination-inhibition antibodies for 9-10 years. These results indicate that the immunity induced by successful primary immunization may persist for at least 15 years. Within this period, a second dose of vaccine only induces low antibody responses which decrease rapidly to their original levels. This provides strong evidence that the immunity produced by primary immunization is long-lasting. However, there were some indications that reimmunization might produce better effects if live attenuated measles virus were used with a longer interval between doses.

PIP: The duration of immunity following measles vaccination of 2882 immunized children has been investigated in a closed region of China for 15 years. A total of 1002 children were treated as primary immunization subjects, and 1547 as reimmunization subjects. These 2 cohorts had no contact with known wild measles virus over the entire observation period, and the results obtained probably reflected the antibody responses to measles vaccine alone. The remaining 333 vaccines came into contact with wild measles virus, and this allowed for the evaluation of the protective effect of the measles vaccines tested. 4 children experienced very mild clinical measles and 329 experienced subclinical infection, including 12 who had had undetectable hemagglutination-inhibition antibodies for 9-10 years. These results indicate that the immunity induced by successful primary immunization may persist for at least 15 years. Within this period of time, a 2nd dose of vaccine only induces low antibody responses which decreases rapidly to their original levels. This demonstrates strong evidence that the immunity produced by primary immunization is long lasting. However, there were indications that reimmunization might produce better results if live attenuated measles virus were used with a longer interval between doses.

Endoscopic lithotripsy of gastric bezoars using a laser-ignited mini-explosive device.

This paper reports the successful experiences of in vitro and in vivo studies on endoscopic fragmentation of gastric bezoars using a laser-ignited mini-explosive device. This new technique has been used clinically in 31 patients with gastric bezoars since January 1988. All patients were completely cured without any complication. The relationship between gastric bezoars and ulcers, and the technique of treatment were discussed.