Chemical and microbiological characterization of two traditional Mongolian high-fat dairy products produced in Xilin Gol, China.

Mongolian butter and Tude are traditional high-fat dairy products produced in Xilin Gol, China, which have unique chemical and microbiological characteristics. Mongolian Tude is made from Mongolian butter, dreg, and flour. In this study, the traditional manufacturing process of Mongolian butter and Tude was investigated for the first time. Mongolian butter was characterized by high-fat content (99.38 ± 0.63%) and high acidity (77.09 ± 52.91°T), whereas Mongolian Tude was considered a high-fat (21.45 ± 1.23%) and high-protein (8.28 ± 0.65%) dairy product obtained by butter, dreg, and flour. Mongolian butter and Tude were proven to be safe for human consumption in terms of benzopyrene content. In addition, Listeria monocytogenes, Staphylococcus aureus, Salmonella, coliforms, and aflatoxin M1 were not detected in the samples. Bacteria and molds were not isolated from Mongolian butter; in contrast, the total count of bacteria and molds in Mongolian Tude was within the range of 4.5 × 102 to 9.5 × 104 and 0 to 2.2 × 105, respectively. Moreover, Lactococcus (41.55%), Lactobacillus (11.05%), Zygosaccharomyces (40.20%), and Pichia (12.90%) were the predominant bacterial and fungal genera, and Lactobacillus helveticus (15.6%), Lactococcus raffinolactis (9.6%), Streptococcus salivarius (8.5%), Pantoea vagans (6.1%), Bacillus subtilis (4.2%), Kocuria rhizophila (3.5%), Acinetobacter johnsonii (3.5%), Zygosaccharomyces rouxii (46.2%), Pichia fermentans (14.7%), and Dipodascus geotrichum (11.7%) were the predominant species in the microbiota of Mongolian Tude. Thus, it can be stated that the microbiota of food products produced by different small families varied significantly. Collectively, the findings presented herein are the first report of chemical and microbiological characterization of products of geographical origin and highlight the need for standardization of manufacturing procedures of Mongolian butter and Tude in the future.

Determination of the microbial community of traditional Mongolian cheese by using culture-dependent and independent methods.

Mongolian cheese is not only a requisite source of food for the nomadic Mongolian but also follows a unique Mongolian dairy artisanal method of production, possessing high nutritional value and long shelf-life. In this study, the ancient technique for the production of Mongolian cheese was investigated. The nutritional value of Mongolian cheese was characterized by its high-protein content (30.13 ± 2.99%) and low-fat content (9.66 ± 3.36%). Lactobacillus, Lactococcus, and Dipodascus were the predominant bacterial and fungal genera, and Lactobacillus helveticus, Lactococcus piscium, and Dipodascus geotrichum were the predominant species in the Mongolian cheese. The microbiota of products from different cheese factories varies significantly. The high-temperature (85°C-90°C) kneading of coagulated curds could eliminate most of the thermosensitive microorganisms for extending the shelf-life of cheese. The indigenous spore-forming microbes, which included yeasts, belonging to Pichia and Candida genera, and molds, belonging to Mucor and Penicillium genera, which originated from the surroundings during the process of cooling, drying, demolding, and vacuum packaging could survive and cause the package to swell and the cheese to grow mold. The investigation of production technology, nutrition, microbiota, and viable microbes related to shelf-life contributes to the protection of traditional technologies, extraction of highlights (nutritional profiles and curd scalding) for merchandise marketing, and standardization of Mongolian cheese production, including culture starters and aseptic technique.

Identifying FecB genotypes in the muscle from sheep breeds indigenous to Xilingol, and establishment of a TaqMan real-time PCR technique to distinguish FecB alleles.

The muscle from Xilingol indigenous sheep breeds are famous in China, and the FecB genotype in this population remains uncharacterized. In this study, SNPs in the FecB locus were investigated by pyrosequencing, and an optimized PCR-RFLP technique was generated to identify SNPs. In addition, an efficient technique for high-throughput identification of SNPs in FecB was optimized using TaqMan real-time PCR and breed-conservative primers and SNP-specific probes. By genotyping the FecB locus in the muscle of Xilingol indigenous sheep breeds using a novel TaqMan real-time PCR assay, our study has generated the groundwork for the authentication of Xilingol mutton based on the specific gene and the prolificacy-oriented breeding of Xilingol sheep using marker-assisted selection strategies in the future.

Improved triplex real-time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat.

The authentication and labeling of meat products, concerning origins and species, are key to fair trade and to protect consumer interests in the market. We developed an improved triplex real-time PCR approach to simultaneously identify chicken, duck, and goose DNA in meat, including an endogenous control to avoid false negatives. Our method specifically detected DNA from chicken, duck, and goose, and showed no cross-reaction with DNA extracted from other meat types. The detection limits of chicken, duck, and goose DNA were 0.001-0.00025 ng, 0.0025-0.0001 ng, and 0.001-0.00001 ng, respectively, and we were able to simultaneously identify DNA from two distinct origins using as little as 0.1% of total meat weight. Our newly generated triplex real-time PCR method with endogenous control exhibited high specificity, sensitivity, and efficiency for simultaneous identification of DNA from chicken, duck, and goose in meat.

A snapshot study of the microbial community dynamics in naturally fermented cow's milk.

Natural fermentation of milk is a prerequisite in the production of traditional dairy products and is considered a bioresource of fermentative microorganisms and probiotics. To understand the microbial dynamics during distinct fermentative phases, the roles of different microbes, and the relationship between bacteria and fungi, microbial community dynamics was investigated by culture-dependent and culture-independent approaches. Natural, static fermentation of milk induces the formation of the underlying curds and the superficial sour cream (Zuohe in the Mongolian language). From an overall perspective, viable LAB increased remarkably. Yeast showed an initial increase in their abundance (from 0 hr to 24 hr), which was followed by a decrease, and mold was detected at the later stages of fermentation (after 68 hr). The observed trends in microbiota variation suggest an antagonistic interaction between bacteria (LAB) and fungi (yeast and mold). The beneficial bacterial and fungal genus and species (e.g., Lactococcus, Streptococcus, Leuconostoc, Dipodascus, Lactococcus lacti, Dipodascus australiensis) are gradually increased in concentration, and the potentially detrimental microbial genus and species (e.g., Acinetobacter, Pseudomonas, Fusarium, Aspergillus, Mortierella, Acinetobacter johnsonii, Fusarium solani) decrease during the decline of bacterial and fungi diversity from natural fermentation. The study of microbial community dynamics could make a great contribution to understand the mechanism of natural fermentation of milk and the formation of curds and Zuohe, and to discover the potentially fermentative microbes for industrial starter cultures.

Comparative study of physicochemical composition and microbial community of Khoormog, Chigee, and Airag, traditionally fermented dairy products from Xilin Gol in China.

Due to their outstanding nutritional and functional properties, the traditionally fermented dairy products (TFDP) from camel, mare, and cow gained universal praise during their long history of production. In this study, the physicochemical composition and microbial communities of Khoormog, Chigee, and Airag, the TFDP from Xilin Gol in China, were investigated and compared. The physicochemical analysis revealed a higher content of total solid content, protein, and fat in Khoormog (12.5 ± 1.6%; 4.6 ± 0.7%; 4.4 ± 1.3%) compared to Chigee (7.8 ± 1.3%; 2.1 ± 0.2%; 0.8 ± 0.2%) and Airag (8.9 ± 0.7%; 3.7 ± 0.4%; 1.4 ± 0.5%). All three types of TFDP shared 41.2% of bacterial and 25.4% of fungal OTUs, and 95.34% of bacterial and 95.52% of fungal sequence reads. The bacterial and fungal community consisted of four phyla and five genera, and three phyla and seven genera, respectively. Lastly, Lactobacillus predominated in Khoormog, Chigee, and Airag at the genus level, while the dominant fungal genera varied among the samples. In conclusion, the microbial community structures of TFDP from camel, mare, and cow were not significantly different in a definite area (Xilingol region), and Khoormog, Chigee, and Airag bred the common "core microbiota".

Simultaneous detection of ovine and caprine DNA in meat and dairy products using triplex TaqMan real-time PCR.

In this study, we report a new approach for the detection of ovine and caprine DNA in meat and dairy products using real-time PCR protocol. Our new approach is based on the use of endogenous control and species-specific TaqMan fluorescence probes. With this methodology, we specifically detected ovine and caprine DNA in meat and dairy products, with limits of detection of 0.001 ng and 0.01 ng for fresh and processed ovine meats, respectively, and 0.00025 ng, 0.005 ng, and 0.01 ng for caprine meat, milk, and cheese, respectively. Artificial meat and milk mixtures from sheep and goat were used to validate the protocol. Our results support that TaqMan real-time PCR with endogenous control is an efficient and accurate method to detect DNA from sheep and goat in meat and dairy products.

Production technology, nutritional, and microbiological investigation of traditionally fermented mare milk (Chigee) from Xilin Gol in China.

Mare milk originated from female horses, known as mares, to feed their foals during lactation. The health-promoting characteristics of traditionally fermented mare milk (Chigee) are well known for the function of clinic treatment in the traditional Mongolian medicine. This study was conducted to investigate the production technology of Chigee and to evaluate the nutritional and microbiological characteristics of mare milk and Chigee based on 188 samples. The nutritional analysis of mare milk and Chigee indicated that lactose significantly decreased from 6.95 ± 0.45% to 2.82 ± 1.65% and acidity and alcoholic content significantly increased to 136.72 ± 57.88°T and 1.22 ± 0.7%, respectively, after spontaneous fermentation of mare milk. The microbiological analysis of Chigee showed that the total lactic acid bacteria (LAB) count varied from 5.32 to 8.56 log cfu/ml and total yeast count varied from 2.41 to 6.98 log cfu/ml. Moreover, the acidity of Chigee rose with the increase in LAB count within limits, and high acidity (≥178°T) inhibited the growth of coliforms. These findings provide an understanding of traditional production technology, nutrition, and microbiology that is fundamental for establishing the food standard of Chigee in China and will contribute to standardize the fermentation process for the industrial production of Chigee in the future.

Investigation of physicochemical composition and microbial communities in traditionally fermented vrum from Inner Mongolia.

Mongolian traditionally fermented vrum is known for its functional characteristics, and indigenous microbial flora plays a critical role in its natural fermentation. However, studies of traditionally fermented vrum are still rare. In this study, we investigated the artisanal production of traditionally fermented vrum from Inner Mongolia. In general, its physicochemical composition was characterized by 34.5 ± 8% moisture, 44.9 ± 12.1% fat, 10.6 ± 3.2% protein, and 210 ± 102°T. The total lactic acid bacteria and yeast counts ranged from 50 to 2.8 × 108 cfu/g and from 0 to 1.1 × 106 cfu/g, respectively. We studied bacterial and fungal community structures in 9 fermented vrum; we identified 5 bacterial phyla represented by 11 genera (an average relative abundance >1%) and 8 species (>1%), and 3 fungal phyla represented by 8 genera (>1%) and 8 species (>1%). Relative abundance values showed that Lactococcus and Lactobacillus were the most common bacterial genera, and Dipodascus was the predominant fungal genus. This scientific investigation of the nutritional components, microbial counts, and community profiles in Mongolian traditionally fermented vrum could help to develop future functional biomaterials and probiotics.

Study of bacterial and fungal community structures in traditional koumiss from Inner Mongolia.

Koumiss is notable for its nutritional functions, and microorganisms in koumiss determine its versatility. In this study, the bacterial and fungal community structures in traditional koumiss from Inner Mongolia, China, were investigated. Our results demonstrated that 6 bacterial phyla represented by 126 genera and 49 species and 3 fungal phyla represented by 59 genera and 57 species were detected in 11 samples of artisanal koumiss. Among them, Lactobacillus was the predominant genus of bacterium, and Kluyveromyces and Saccharomyces dominated at the fungal genus level. In addition, there were no differences in the bacterial and fungal richness and diversity of koumiss from 3 neighboring administrative divisions in Inner Mongolia, and the bacterial and fungal community structures (the varieties and relative abundance of bacterial and fungal genera and species) were clearly distinct in individual samples. This study provides a comprehensive understanding of the bacterial and fungal population profiles and the predominant genus and species, which would be beneficial for screening, isolation, and culture of potential probiotics to simulate traditional fermentation of koumiss for industrial and standardized production in the future.

Optical radiation stability of ZnO hollow particles.

Zinc oxide has multifunctional physical properties depending on its microstructure and morphology. Herein, we reported the in situ investigations of the radiation stability of ZnO particles with hollow, ball, star and flower shapes under electron and proton irradiation. 100 keV protons with a fluence of 5 × 1015 cm-2 and 50 keV electrons with fluence ranging from 0.5 to 7 × 1016 cm-2 are employed to investigate the radiation stability of nanostructured ZnO particles. In situ reflectance, X-ray photoelectron spectra and photoluminescence were characterized in the irradiation environment to avoid the effects of the atmospheric environment on radiation induced defects. The experimental results reveal that, compared to the other shapes, the hollow structure with the best radiation stability due to the hollow structure facilitates the decrease of the accumulation of radiation defects. This study clearly demonstrates the promise of ZnO hollow particles as a plasmonic nanostructure for achieving high radiation stability, and they could be easily employed to serve as the radiation stability pigment for coatings.

Fish oil supplementation improves pregnancy outcomes and size of the newborn: a meta-analysis of 21 randomized controlled trials.

OBJECTIVE: To observe the effects of fish oil on related pregnancy outcomes. METHODS: A systematic search of the Medline, EMBASE and Cochrane's library databases was conducted for the randomized controlled trials published till February 2015 that compared the effects of fish oil supplementation with a control diet in women during pregnancy. RESULTS: Twenty-one studies comprising 10,802 pregnant women were included. Dietary fish oil was associated with a 5.8-day increase in gestational age of the newborn, a 22% reduced risk for early preterm delivery (risk ratio [RR] = 0.78, 95% CI: 0.64-0.95, p = 0.01) and a 10% reduction in preterm delivery (RR = 0.90, 95% CI 0.81-1.00, p = 0.05). Fish oil supplementation was associated with higher infantile birth weight (51.23 g), birth length (0.28 cm) and head circumference (0.09 cm), and a 23% lower risk of low birth weight. No benefit from fish oil supplementation was found with regard to risk of intrauterine growth restriction or stillbirth. CONCLUSIONS: Dietary fish oil during pregnancy was associated with reduced risk of preterm delivery and improved size of the newborn. Fish oil during pregnancy may be an effective prophylactic for preterm delivery.

Screening of biomarkers in cervical squamous cell carcinomas via gene expression profiling.

In the present study, gene expression profiles of high-grade squamous intraepithelial lesions (HSIL) and invasive cervical squamous cell carcinomas (CSCC) were analyzed using bioinformatic tools to identify key genes and potential biomarkers. Analyses of differentially expressed genes (DEGs) were performed for HSIL vs. normal control and invasive CSCC vs. normal control tissues using the Limma package in R. Pathway enrichment analysis was performed using KOBAS. A protein­protein interaction (PPI) network for the DEGs in invasive CSCC was constructed using String. Functional enrichment analysis was performed for the DEGs in the PPI network using DAVID. Relevant small molecules were predicted using Cmap. A total of 633 and 881 DEGs were identified in HSIL and invasive CSCC, respectively, and the two groups had 305 DEGs in common. Genes associated with the mitogen-activated protein kinase signaling pathway were enriched in the HSIL, while cell cycle-associated genes were over­represented in invasive CSCC. The PPI network, containing 72 upregulated genes and 434 edges, was illustrated. Functional enrichment analysis showed that the cell cycle was the most significant gene ontology term. A total of six small molecules associated with the pathology of CSCC were identified, including the anti-cancer drug piperlongumine, which showed a negative correlation. The findings of the present study not only enhanced the current understanding of the pathogenesis of CSCC, but may also be a basis for the development of novel therapies.

Fish Oil Supplementation does not Reduce Risks of Gestational Diabetes Mellitus, Pregnancy-Induced Hypertension, or Pre-Eclampsia: A Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: The effects of gestational supplementation with fish oil on risks for gestational diabetes mellitus (GDM), pregnancy-induced hypertension (PIH), and pre-eclampsia (PE) have not been confirmed. In this study, a meta-analysis was performed to evaluate the effect of fish oil supplementation on these gestational complications. MATERIAL AND METHODS: Randomized controlled human trials that investigated the effects of fish oil supplementation in pregnant women were identified by a systematic search of Medline, Embase, and Cochrane's Library, and references of related reviews and studies up to December 2014. Relative risks (RRs) for GDM, PIH, and PE were the outcomes of interest. Fixed-effects or random-effects models were applied according to the heterogeneity. RESULTS: Thirteen comparisons from 11 published articles, including more than 5000 participants, were included. The results showed that fish oil supplementation was not associated with reduced risks for GDM (RR=1.06, 95% confidence interval [CI]: 0.85-1.32, p=0.60), PIH (RR=1.03, 95% CI: 0.89-1.20, p=0.66), or PE (RR=0.93, 95% CI: 0.74-1.16, p=0.51). No statistically significant heterogeneity was detected for the comparison of each outcome. The effects of fish oil on these gestational complications were consistent between women with low-risk and high-risk pregnancies. CONCLUSIONS: Gestational supplementation with fish oil during the second or third trimester of pregnancy is not associated with reduced risks for GDM, PIH, or PE. Other possible benefits of fish oil supplementation during pregnancy warrant further evaluation.

Crocetin downregulates the proinflammatory cytokines in methylcholanthrene-induced rodent tumor model and inhibits COX-2 expression in cervical cancer cells.

The effect of crocetin (C20H24O4) on methylcholanthrene- (MCA-) induced uterine cervical cancer in mice was studied in this paper. After the mice were treated orally with crocetin, maleic dialdehyde (MDA), polymorphonuclear cells (PMN), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α) were examined by ELISA or immunohistochemistry. The inducible nitric oxide synthase (iNOS) activation in HeLa cells was analyzed using fluorescence microscopy for light microscopic examination. The MCA mice showed a significant increase in plasma MDA, PMN, IL-1ß, TNF-α, and nitrates levels. At the same time, the mRNA level of COX-2 in HeLa cells was also significantly increased. These changes were attenuated by crocetin supplementation in the MCA mice. Crocetin supplementation in the MCA mice also showed protection against cervical cancer. These results suggest that crocetin may act as a chemopreventive and an anti-inflammatory agent.

MiRNA-494 inhibits metastasis of cervical cancer through Pttg1.

Many cervical cancer (CC) patients experience early cancer metastasis, resulting in poor therapeutic outcome after resection of primary cancer. Hence, there is a compelling requirement for understanding of the molecular mechanisms underlying the invasiveness control of CC. Pituitary tumor-transforming gene 1 (Pttg1) has been recently reported to promote cancer cell growth and metastasis in a number of various tumors. However, its regulation by microRNAs (miRNAs) as well as its role in CC have not been clarified. Here, we reported significantly higher levels of Pttg1 and significantly lower levels of miR-494 in the resected CC tissue, compared with the adjacent normal cervical tissue from the same patient. Interestingly, Pttg1 levels inversely correlated with miR-494 levels. In vitro, Pttg1 levels determined CC cell invasiveness and were inhibited by miR-494 levels. However, miR-494 levels were not affected by Pttg1 levels. Furthermore, miR-494 inhibited Pttg1 expression in CC cells, through directly binding and inhibition on 3'-UTR of Pttg1 mRNA. Together, our data suggest that Pttg1 may increase CC cell metastasis, which is negatively regulated by miR-494. Our work thus highlights a novel molecular regulatory machinery in metastasis of CC.

Effects of suberoylanilide hydroxamic acid (SAHA) combined with paclitaxel (PTX) on paclitaxel-resistant ovarian cancer cells and insights into the underlying mechanisms.

BACKGROUND: Suberoylanilide hydroxamic acid (SAHA) is a member of the hydroxamic acid class of the newly developed histone deacetylase inhibitors. Recently, Suberoylanilide hydroxamic acid has attracted increasing attention because of its antitumor activity and synergistic effects in combination with a variety of traditional chemotherapeutic drugs. Paclitaxel (PTX), is a natural anticancer drugs; however, resistance to paclitaxel has become a major challenge to the efficacy of this agent. The purpose of this study was to investigate the effects of the combined application of these two drugs on the paclitaxel-resistant ovarian cancer OC3/P cell line. METHODS: In the present study, the effects of Suberoylanilide hydroxamic acid or/and paclitaxel on OC3/P cells cultured in vitro were analyzed in terms of cell viability, migration, cell-cycle progression and apoptosis by CCK-8, wound healing and flow cytometry assays. Changes in cell ultrastructure were observed by transmission electron microscopy. The expression of genes and proteins related to proliferation, apoptosis and drug resistance were analyzed by quantitative real-time polymerase chain reaction and Western blot analyses. RESULTS: There was no cross-resistance of the paclitaxel-resistant ovarian cancer OC3/P cells to Suberoylanilide hydroxamic acid. Suberoylanilide hydroxamic acid combined with paclitaxel significantly inhibited cell growth and reduced the migration of OC3/P cells compared with the effects of Suberoylanilide hydroxamic acid or paclitaxel alone. Q-PCR showed the combination of Suberoylanilide hydroxamic acid and paclitaxel reduced intracellular bcl-2 and c-myc gene expression and increased bax gene expression more distinctly than the application of SAHA or paclitaxel alone. Moreover, the level of mdr1 gene expression in cells treated with Suberoylanilide hydroxamic acid was lower than that of the control group (P <0.05). Western blot analysis showed that Suberoylanilide hydroxamic acid alone or in combination with paclitaxel enhanced caspase-3 protein expression and degraded ID1 protein expression in OC3/P cells. CONCLUSION: Suberoylanilide hydroxamic acid inhibited the growth of paclitaxel-resistant ovarian cancer OC3/P cells and reduced migration by the induction of cell-cycle arrest, apoptosis and autophagy. These observations indicate the possible synergistic antitumor effects of sequential Suberoylanilide hydroxamic acid and paclitaxel treatment.

Expression of metastasis suppressor 1 in cervical carcinoma and the clinical significance.

This study aimed to investigate the expression of metastasis suppressor 1 (MTSS1) in cervical intraepithelial neoplasia (CIN) and malignant cervical tissues, and the role of MTSS1 in carcinogenesis. MTSS1 expression was detected by immunohistochemistry in 147 cervical tissue specimens collected from 30 healthy individuals, 30 patients with cervical CIN I, 30 patients with CIN II-III and 57 patients with cervical cancer. The association between MTSS1 expression and clinicopathological factors was also examined. MTSS1 was found to be positively expressed in 43.33% CIN I cervical tissues, 100% CIN II-III cervical tissues and 100% malignant cervical tissues, but was weakly or negatively expressed in benign cervical tissues. The positive expression rates of MTSS1 were significantly higher in CIN II-III and malignant cervical tissues than in CIN I or normal cervical tissues (P<0.05). When examining MTSS1 expression and clinicopathological factors, the strong positive MTSS1 expression rates in early-stage versus middle- and advanced-stage cervical cancer tissues were 39.13% and 82.35%, respectively. Furthermore, the positive expression rates of MTSS1 were significantly higher in cervical tissues at an advanced clinical stage than those at an early clinical stage (P<0.05). The results suggest that the dysregulation of MTSS1 may be involved in cervical carcinogenesis, and thus MTSS1 may be a novel diagnostic biomarker or therapeutic target in cervical cancer patients.

Comparison of vaginal and abdominal hysterectomy:A prospective non-randomized trial.

OBJECTIVE: To compare outcomes of vaginal and abdominal hysterectomy procedures in women with benign gynaecological diseases. METHODS: This was a prospective study of outcomes of consecutive patients who underwent total vaginal hysterectomy (VH) or abdominal hysterectomy (AH) for benign gynaecological diseases. Patient characteristics before, during, and after the operations were reviewed. Patients were followed up for three months to evaluate postoperative complications. RESULTS: This study included a total of 313 patients. 143 patients underwent AH and 170 patients underwent VH. Baseline characteristics were similar between the two groups. There were no intraoperative complications in either group. Operation time, intraoperative blood loss, first postoperative flatus time, time to out-of-bed activity, mean maximum postoperative body temperature, and duration of fever were all significantly shorter and less severe in the VH group compared with the AH group. In addition, vaginal length in the VH group was significantly shorter than in the AH group. CONCLUSIONS: Vaginal hysterectomy has advantages over AH in the treatment of benign gynaecological diseases, providing greater efficacy and safety with minimal invasiveness.