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Rennet, a complex of enzymes found in the stomachs of ruminants, is an important component for cheese production. In our study, we described that yak chymosin gene recombinant Pichia pastoris strain could serve as a novel source for rennet production. Yaks total RNA was extracted from the abomasum of an unweaned yak. The yak preprochymosin, prochymosin, and chymosin genes from total RNA were isolated using gene specific primers based on cattle chymosin gene sequence respectively and analyzed their expression pattern byreal time-polymerase chain reaction. The result showed that the chymosin gene expression level of the sucking yaks was 11.45 times higher than one of adult yaks and yak chymosin belongs to Bovidae family in phylogenetic analysis. To express each, the preprochymosin, prochymosin, and chymosin genes were ligated into the expression vector pPICZαA, respectively, and were expressed in Pichia pastoris X33. The results showed that all the recombinant clones of P. pastoris containing the preprochymosin, prochymosin or chymosin genes could produce the active form of recombinant chymosin into the culture supernatant. Heterologous expressed prochymosin (14.55 Soxhlet unit/mL) had the highest enzyme activity of the three expressed chymosin enzymes. Therefore, we suggest that the yak chymosin gene recombinant Pichia pastoris strain could provide an alternative source of rennet production.

The stomach lysozymes of ruminants play a paramount role in digestion and offer the chance to probe evolutionary changes in complex organisms on a biochemical basis. In this paper, we focus on the characterization of the catalytic properties of Tibetan ruminant (TR) stomach lysozymes and provide a comparative study of TR stomach lysozymes and nonplateau ruminant stomach lysozymes. The stomach lysozymes were purified with a Carboxymethyl (CM) Sepharose Fast Flow (FF) column and a Bio-Gel P-100 column. The purified stomach lysozymes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization time-of-flight (MALDITOF) analyses. The antimicrobial activity was evaluated using an agar diffusion method, and the pH, ion strength, temperature, pepsin and trypsin effects on enzymatic activities were evaluated by normal biochemistry methods. The stomach lysozymes of TRs (cattle:TC, yak:TY, and sheep:TS) exhibit maximum activity at pH 5 and ionic strengths below 0.02. TC and TS stomach lysozymes are more resistant to higher temperatures than nonplateau lysozymes. Stomach lysozymes of TRs exhibit greater enzymatic activity than nonplateau lysozymes, in the presence of pepsin and trypsin. Stomach lysozymes from TS retain 86% of their initial enzymatic activity against pepsin inactivation. After trypsin treatment, the stomach lysozymal activities of TRs were significantly affected by trypsin inactivation, with the exception of TS, whose enzymatic activities in the presence of trypsin were 5- and 10-fold higher than those of nonplateau lysozymes. The stomach lysozymes of TRs are more resistant to environmental factors such as pH, temperature, pepsin, and trypsin than nonplateau lysozyme C. [ABSTRACT FROM AUTHOR]

Objective. The purpose of the study was to examine the effects of laser acupuncture (LA) at right Neiguan (RPC6)/left Neiguan (LPC6) acupoints on the releases of nitric oxide (NO) in the treated and contralateral/nontreated PC6, compared to the nonacupoint control area. Methods. 24 mW LA at RPC6, LPC6, and nonacupoint in 22 healthy subjects for 40 min: sterilized dialysis tube was taped to the nontreated PC6/nonacupoint during the treatment and immediately taped to the treated and nontreated PC6/nonacupoint after LA removal. NO-scavenging compound was injected into the tube for 40 min to absorb the molecular which was tested by spectrophotometry in a blinded fashion. Results. LA-induced NO releases over PC6 acupoints for the nontreated and treated sides all significantly increased after LA removal, but for the nontreated acupoints they did not change during LA stimulation. LA at RPC6 induced the more release of the NO at contralateral side than stimulating LPC6, but not on nonacupoints. The results suggest that LA-induced NO release over contralateral acupoint and NO release resulting from the lateralized specificity all are different and specific to the acupoint within different time course. Conclusions. LA-evoked NO release over acupoints could improve the neurogenic, endothelial activity of the vessel wall to further facilitate microcirculation. [ABSTRACT FROM AUTHOR]

Objectives The aim of this study was to detect the influences of LA at nonacupoint and two adjacent acupoints of pericardium meridian on the releases of NO and sGC in 20 healthy subjects. Methods Different intensities (12, 24, 48 mW) of infrared laser were used for irradiating Jianshi ( PC5), Ximen ( PC4) acupoints and nonacupoint for 20, 40 minutes, respectively. Semi-circular tubes were taped to the skin surface and filled with NO-scavenging compound for 20 minutes to capture NO and sGC, which were measured using spectrophotometry in a blinded fashion. Results As the increase in the intensity of LA stimulation, the levels of NO releases over acupoints all were significantly increased, NO releases in nonacupoints following the same treatment only changed slightly, sGC amounts were observably enhanced over acupoints, but did not any change in nonacupoint area. Different intensities of LA treatments can sensitively affect the NO and sGC releases over acupoints. This indicated that LA-induced releases of the NO and sGC were specific to acupoints. Conclusions This is the first evidence reporting that LA induced significant elevations of NO- sGC releases over acupoints, and the enhanced signal molecules contribute to local circulation, which improves the beneficial effects of the therapy. [ABSTRACT FROM AUTHOR]

Purpose: The prognostic significance of circulating tumor cells (CTCs) in esophageal carcinoma (EC) is controversial. We aim to assess its association with clinicopathological and prognostic relevance in EC by using a meta-analysis. Methods: We searched PubMed, Cochrane Database, Embase databases, and the references in relevant studies that assessed the clinicopathological or prognostic relevance of CTCs in peripheral blood of patients with EC. Statistical analyses were conducted by using Stata software to calculate the pooled odds ratio (OR), hazard ratio (HR), and 95% confidence intervals (CIs) using fixed or random-effects models according to the heterogeneity of included studies. The subgroup analyses were performed according to ethnicity, histological type, and detection method. Results: Sixteen trials containing 1,260 patients were included for analysis. Pooled results showed that presence of CTCs was significantly associated with poor overall survival (HR =1.71, 95% CI [1.30, 2.12], P<0.001) and progression-free survival (HR =1.67, 95% CI [1.19, 2.15], P<0.001) in EC patients. Subgroup analysis indicated that presence of CTCs was closely associated with worse overall survival (Asian: HR =1.66, 95% CI [1.24, 2.08], P<0.001; squamous cell carcinoma [SCC]: HR =1.66, 95% CI [1.24, 2.08], P<0.001; no polymerase chain reaction [PCR]: HR =2.08, 95% CI [1.40, 2.76], P<0.001) and progression-free survival (Asian: HR =1.63, 95% CI [1.15, 2.12], P<0.001; SCC: HR =1.63, 95% CI [1.15, 2.12], P<0.001; PCR: HR =1.63, 95% CI [1.15, 2.12], P<0.001). Additionally, ORs showed that presence of CTCs was significantly correlated with tumor node metastasis (TNM) staging (overall: OR = 1.96, 95% CI [1.34, 2.87], P=0.001; Asian: OR =2.09, 95% CI [1.37, 3.19], P=0.001; SCC: OR =1.97, 95% CI [1.21, 3.07], P=0.003; PCR: OR =2.23, 95% CI [1.43, 3.47], P<0.001), venous invasion (overall: OR =2.23, 95% CI [1.46, 3.40], P<0.001; Asian: OR =2.23, 95% CI [1.46, 3.40], P<0.001; SCC: OR =2.23, 95% CI [1.46, 3.40], P<0.001; PCR: OR =2.23, 95% CI [1.46, 3.40], P<0.001), lymph node metastasis (overall: OR =2.41, 95% CI [1.50, 3.86], P<0.001; Asian: OR =2.89, 95% CI [1.80, 4.65], P<0.001; SCC: OR =2.44, 95% CI [1.47, 4.07], P=0.001; PCR: OR =2.89, 95% CI [1.80, 4.65], P<0.001) and distant metastasis (Asian: OR =2.68, 95% CI [1.01, 7.08], P=0.047) in patients with EC. Conclusion: The presence of CTCs indicates a poor prognosis in EC patients, especially in Asian and SCC patients. Further well-designed prospective studies are recommended to explore the clinical applications of CTCs in patients with EC. [ABSTRACT FROM AUTHOR]

Background: Germline mutations in PALB2 gene make a small contribution to heritable breast cancer susceptibility. A recent report has revealed that women with mutations in the PALB2 gene were more than nine times as likely to develop breast cancer compared to those without. The aim of this study is to understand the status of PALB2 mutations among Chinese high-risk breast cancer patients in a multi-ethnic region in China. Methods: 152 patients with hereditary predisposition to breast cancer from the Xinjing region of China were enrolled in the study, and 100 control samples from healthy women were collected in the same locality. We sequenced the coding sequences and flanking intronic regions of PALB2 gene from DNA samples obtained from all subjects by direct sequencing. Results: A total of 4 deleterious PALB2 mutations were identified in 152 breast cancer patients with a prevalence of about 2.6% (4/152). The PALB2 mutation prevalence was 3.2% (3/95) in cases with family history of breast cancer. In addition to the four deleterious mutations, we identified nine missense variants in 12 patients, using the prediction Softwares SIFT and PolyPhen, four of which might be disease associated (in 5 patients). Two of the 4 patients with deleterious mutations and 2 of the 5 patients presenting putative deleterious missense mutations had triple-negative breast cancer. No PALB2 mutation carriers were identified in 100 healthy controls. Conclusion: PALB2 mutations account for a small, but not negligible, proportion of patients with hereditary predisposition to breast cancer in the Xinjing region of China. [ABSTRACT FROM AUTHOR]

The Colorado potato beetle (Leptinotarsa decemlineata (Say)) in the north Xinjiang Uygur autonomous region has evolved resistance to various types of insecticides. Chlorantraniliprole is a novel anthranilic diamide insecticide that binds and activates ryanodine receptors. It exhibited excellent efficacy against L. decemlineata in several field trails in Europe. In the present paper, the susceptibility of L. decemlineata fourth-instar larvae derived from six field populations and L. decemlineata adults derived from three field populations to chlorantraniliprole was determined by a topical application. The fourth-instar larvae were substantially more susceptible to chlorantraniliprole than adults, although the range of susceptibility was far greater among the fourth-instar larvae. Regarding stomach toxicities, adult beetles were less susceptible to chlorantraniliprole than larvae. Chlorantraniliprole was most toxic to second-instar larvae, followed by third- and fourth-instar larvae. These data suggested that the appropriate timing for chlorantraniliprole spraying is the early larval stage. Moreover, the synergistic activities of chlorantraniliprole in combination with triphenyl phosphate, diethyl maleate, or piperonyl butoxide against fourth-instar larvae from two field populations and adults from one field population were tested. Piperonyl butoxide had synergistic effects with chlorantraniliprole against fourth-instar larvae but not against adult beetles. Conversely, triphenyl phosphate and diethyl maleate exerted little synergistic effects. It appears that there is a potential risk of resistance against chlorantraniliprole resulting from cytochrome P450 monooxygenase activity. [ABSTRACT FROM AUTHOR]

Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), has become the economically most important insect defoliator of potatoes, Solanum tuberosum L., in northern Xinjiang Uygur autonomous region in China. Currently, control of Colorado potato beetle relies mainly on chemical insecticides. And this may result in insecticide resistance. In this study, LDm values were measured by a topical bioassay for 14 conventional insecticides in seven local populations from Urumqi, Changji, Tacheng, Nilka, Gongliu, Qapqal, and Tekes counties (cities). The Tekes field population was the most susceptible population and was selected as a reference strain. Compared with the Tekes strain, the Changji, Qapqal, Nilka, Tacheng, and Congliu populations exhibited moderate to very high levels of resistance to cyhalothrin. The Qapqal and Changji populations showed a moderate and a very high resistance to deltarnethrin, respectively. And the Changji population developed a high resistance against a-cypermethrin. Moreover, the Qapqal population had a moderate resistance to carbofuran, and the Urumqi population reached high level of resistance to endosulfan. Possible resistance mechanisms of the Changji and Qapqal populations were determined using three enzyme inhibitors. Triphenyl phosphate (TPP), diethylmeleate, and piperonyl butoxide (PBO) had little synergism to cyhalothrin in the two populations. In contrast, PBO and TPP exhibited some synergistic effects to carbofuran in the Qapqal population, indicating the involvement of monooxygenases and esterases in conferring carbofuran resistance. It seems that additional mechanisms, such as target site insensitivity, should play an important role in Colorado potato beetle resistances to cyhalothrin and carbofuran in northern Xinjiang local populations. [ABSTRACT FROM AUTHOR]

The existence of solutions for the 2n-order m-point boundary value problem at resonance is obtained by using the coincidence degree theory of Mawhin. We give an example to demonstrate our result. The interest is that the nonlinear term may be noncontinuous. [ABSTRACT FROM AUTHOR]

Background: The purpose of the study is to determine the optical properties and their differences for normal human stomach mucosa/submucosa tissue in the cardiac orifice in vitro at 635, 730, 808, 890 and 980 nm wavelengths of laser.Methods: The measurements were performed using a CCD detector, and the optical properties were assessed from the measurements using the spatially resolved reflectance, and nonlinear fitting of diffusion equation.Results: The results of measurement showed that the absorption coefficients, the reduced scattering coefficients, the optical penetration depths, the diffusion coefficients, the diffuse reflectance and the shifts of diffuse reflectance of tissue samples at five different wavelengths vary with a change of wavelength. The maximum absorption coefficient for tissue samples is 0.265 mm-1 at 980 nm, and the minimum absorption coefficient is 0.0332 mm-1 at 730 nm, and the maximum difference in the absorption coefficients is 698% between 730 and 980 nm, and the minimum difference is 1.61% between 635 and 808 nm. The maximum reduced scattering coefficient for tissue samples is 1.19 mm-1 at 635 nm, and the minimum reduced scattering coefficient is 0.521 mm-1 at 980 nm, and the maximum difference in the reduced scattering coefficients is 128% between 635 and 980 nm, and the minimum difference is 1.15% between 890 and 980 nm. The maximum optical penetration depth for tissue samples is 3.57 mm at 808 nm, and the minimum optical penetration depth is 1.43 mm at 980 nm. The maximum diffusion constant for tissue samples is 0.608 mm at 890 nm, and the minimum diffusion constant is 0.278 mm at 635 nm. The maximum diffuse reflectance is 3.57 mm-1 at 808 nm, and the minimum diffuse reflectance is 1.43 mm-1 at 980 nm. The maximum shift Deltax of diffuse reflectance is 1.11 mm-1 at 890 nm, and the minimum shift Deltax of diffuse reflectance is 0.507 mm-1 at 635 nm.Conclusion: The absorption coefficients, the reduced scattering coefficients, the optical penetration depths, the diffusion coefficients, the diffuse reflectance and the shifts of diffuse reflectance of tissue samples at 635, 730, 808, 890 and 980 nm wavelengths vary with a change of wavelength. There were significant differences in the optical properties for tissue samples at five different wavelengths (P < 0.01). [ABSTRACT FROM AUTHOR]

Transcription factors (TFs) are crucial components of regulatory networks that control gene transcription. Current TF assays are limited to the analysis of a single TF or require TF-specific antibodies. Here we report the Single Primer Amplification assisted Oligonucleotide Array-based Transcription Factor Assay (SPA-OATFA) which can directly analyze the binding activities of 240 human TFs simultaneously. Examining early events during serum-stimulation of HeLa cells as a model, we demonstrated the utility of SPA-OATFA combined with whole genome gene expression to systematically map the temporal activation of signaling pathways. Both TFs known to function in this stimulation response such as EGR1 and AP1 and new TFs such as HSF1 were identified. This information, combined with mRNA profiling, provided novel insights into the activities of regulatory pathways, and illustrates the potential of SPA-OATFA in detailed systems biology analysis of cell responses.

Cell/tissue-specific gene expression are tightly regulated by various combinations of multiple transcription factors (TFs). Here, we present an oligonucleotide array-based transcription factor assay (OATFA), which allows the simultaneous assay of multiple active TFs. In this proof-of-principle work, both purified TFs and cell extracts were analyzed using OATFA and further antibody-based validation confirmed the chip data. This method could simplify the assay of multiple TFs and may facilitate high-throughput profiling of large numbers of TFs.

The purpose of the study is to analyze and compare differences in the optical properties between normal and adenomatous human colon tissues in vitro at 630-, 680-, 720-, 780-, 850-, and 890-nm wavelengths using a Ti:sapphire laser. The optical parameters of tissue samples are determined using a double integrating sphere setup at seven different laser wavelengths. The inverse Monte Carlo simulation is used to determine the optical properties from the measurements. The results of measurement show that the optical properties and their differences vary with a change of laser wavelength for normal and adenomatous colon mucosa/submucosa and normal and adenomatous colon muscle layer/chorion. The maximum absorption coefficients for normal and adenomatous human colon mucosa/submucosa are 680 nm, and the minimum absorption coefficients for both are 890 nm. The maximum difference of the absorption coefficients between both is 56.8% at 780 nm. The maximum scattering coefficients for normal and adenomatous colon mucosa/submucosa are 890 nm, and the minimum scattering coefficients for both are 780 nm. The maximum difference of the scattering coefficients between both is 10.6% at 780 nm. The maximum absorption coefficients for normal and adenomatous colon muscle layer/chorion are 680 nm, and the minimum absorption coefficients for both are 890 nm. The maximum difference of the absorption coefficients between both is 47.9% at 780 nm. The maximum scattering coefficients for normal and adenomatous colon muscle layer/chorion are 890 nm, and the minimum scattering coefficients for both are 680 nm. The maximum difference of the scattering coefficients between both is 9.61% at 850 nm. The differences in absorption coefficients between normal and adenomatous tissues are more significant than those in scattering coefficients.

Aim: The purpose of the present study is to compare the optical properties of normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion in vitro at 476.5, 488, 496.5, 514.5 and 532 nm. We believe these differences in optical properties should help differential diagnosis of human colon tissues by using optical methods., Methods: In vitro optical properties were investigated for four kinds of tissues: normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion. Tissue samples were taken from 13 human colons (13 adenomatous, 13 normal). From the normal human colons a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion), and from the adenomatous human bladders a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion). The measurements were performed using a double-integrating-sphere setup and the optical properties were assessed from these measurements using the adding-doubling method that was considered reliable., Results: The results of measurement showed that there were significant differences in the absorption coefficients and scattering coefficients between normal and adenomatous human colon mucosa/submucosa at the same wavelength, and there were also significant differences in the two optical parameters between both colon muscle layer/chorion at the same wavelength. And there were large differences in the anisotropy factors between both colon mucosa/submucosa at the same wavelength, there were also large differences in the anisotropy factors between both colon muscle layer/chorion at the same wavelength. There were large differences in the value ranges of the absorption coefficients, scattering coefficients and anisotropy factors between both colon mucosa/submucosa, and there were also large differences in these value ranges between both colon muscle layer/chorion. There are the same orders of magnitude in the absorption coefficients for four kinds of colon tissues. The scattering coefficients of these tissues exceed the absorption coefficients by at least two orders of magnitude., Conclusion: There were large differences in the three optical parameters between normal and adenomatous human colon mucosa/submucosa at the same laser wavelength, and there were also large differences in these parameters between both colon muscle layer/chorion at the same laser wavelength. Large differences in optical parameters indicate that there were large differences in compositions and structures between both colon mucosa/submucosa, and between both colon muscle layer/chorion. Optical parameters for four kinds of colon tissues are wavelength dependent, and these differences would be useful and helpful in clinical applications of laser and tumors photodynamic therapy (PDT).

Aim: To study the optical properties of human normal small intestine tissue at 476.5 nm, 488 nm, 496.5 nm, 514.5 nm, 532 nm, 808 nm wavelengths of laser irradiation., Methods: A double-integrating-sphere system, the basic principle of measuring technology of light radiation, and an optical model of biological tissues were used in the study., Results: The results of measurement showed that there were no significant differences in the absorption coefficients of human normal small intestine tissue at 476.5 nm, 488 nm, 496.5 nm laser in the Kubelka-Munk two-flux model (P>0.05). The absorption coefficients of the tissue at 514.5 nm, 532 nm, 808 nm laser irradiation were obviously increased with the decrease of these wavelengths. The scattering coefficients of the tissue at 476.5 nm, 488 nm, 496.5 nm laser irradiation were increased with the decrease of these wavelengths. The scattering coefficients at 496.5 nm, 514.5 nm, 532 nm laser irradiation were obviously increased with the increase of these wavelengths. The scattering coefficient of the tissue at 532 nm laser irradiation was bigger than that at 808 nm. There were no significant differences in the total attenuation coefficient of the tissue at 476.5 nm and 488 nm laser irradiation (P>0.05). The total attenuation coefficient of the tissue at 488 nm, 496.5 nm, 514.5 nm, 532 nm, 808 nm laser irradiation was obviously increased with the decrease of these wavelengths, and their effective attenuation coefficient revealed the same trend. There were no significant differences among the forward scattered photon fluxe, backward scattered photon fluxe, and total scattered photon fluxe of the tissue at 476.5 nm, 488 nm, 496.5 nm laser irradiation. They were all obviously increased with attenuation of tissue thickness. The attenuations of forward and backward scattered photon fluxes, and the total scattered photon fluxe of the tissue at 514.5 nm laser irradiation were slower than those at 476.5 nm, 488 nm, 496.5 nm laser irradiation respectively. The attenuations of forward and backward scattered photon fluxes, and total scattered photon fluxes at 532 nm laser irradiation were obviously slower than those at 476.5 nm, 488 nm, 496.5 nm, 514.5 nm laser irradiation. The attenuations of forward and backward scattered photon fluxes, and total scattered photon fluxe at 808 nm laser irradiation were all obviously slower than those at 476.5 nm, 488 nm, 496.5 nm, 514.5 nm, 532 nm laser irradiation respectively., Conclusion: There are significant differences in optical parameters of human normal small intestine tissue in the Kubelka-Munk two-flux model at six different wavelengths of laser radiation. The results would provide a new method of information analysis for clinical diagnosis.