Cancer-associated fibroblasts subtypes and role in invasion and metastasis of gastric cancer.

Gastric cancer (GC) is the fifth most common malignancy and the fourth leading cause of cancer-related death worldwide. Cancer-associated fibroblasts (CAFs), an important cell type in the tumor microenvironment, play an important role in GC development. In this review, we describe the current knowledge of CAFs' heterogeneity and their role in GC invasion and metastasis. Currently, CAF-targeted cancer therapies are being rapidly explored and developed. However, the heterogeneity of CAFs limits the application of this therapy, so it is urgent to find specific markers and divide them into different subpopulations. With the development of single-cell RNA sequencing technology, researchers have used this technology to classify CAFs in many tumors, but whether it is applicable to GC and other tumors needs further study. And we believe that this technology will be in the near future utilized to sort CAFs on the basis of different cell markers and functions, so as to target tumor-promoting CAFs and inhibit tumor progression. Targeting CAFs by cell surface markers or normalizing the activated CAFs subsets may be an effective therapy, alone or in combination with other therapeutic approaches for GC treatment. Therefore, in the coming decades, the interaction between CAFs and GC cells will be still the focus of our research.

Effects of water-immersion cooling temperatures on the moisture retention of sodium-reduced pork sausages.

To improve the industrial yield of sodium-reduced meat products, we present a feasible method by adjusting water-immersion cooling temperatures to decrease the water loss of pork sausage during processing. The present results showed that the moisture retention capacity of sodium-reduced pork sausages (SRPS) cooled by the temperatures of 15-20 °C was larger than that of 0-10 °C. The higher cooling temperatures, especially at 20 °C, could change the movement and population of proton of inner water, decrease syneresis and facilitate the formation of homogenous cross-linked network, thus increasing the moisture retention of SRPS. Results of this work indicated that the control of cooling temperature of sodium-reduced sausages after cooking could serve as a feasible approach for improving the economic benefits and quality characteristics of the final products.

[Real-time dynamic recording of cerebral cortical vascular embolization and regeneration in rats].

The purpose of this study was to establish a method to record the dynamic process of vascular regeneration and remodeling in rat cerebral ischemic regions. An animal brain window model was established to continuously observe the changes of rat cortical vascular ischemia in vivo, and the model of cerebral ischemia was established by photochemical embolization. Optical coherence tomography (OCT) was performed to record the formation of vascular blockage and the injury and regeneration of small vessels during cerebral ischemia recovery. The results showed that 30 min of laser irradiation could completely block the cortical vessels in rats. Within 24-48 h after ischemia, the degree of brain injury was the greatest, and the number of blood vessels in the ischemic region reached the minimum. Then the blocked blood vessels began to be dredged, and the small blood vessels around the ischemic area began to regenerate. Small blood vessels in the superficial/deep layers of the cortex disappeared significantly after laser irradiation. During 10 d after ischemia, the blocked blood vessels were gradually dredged and recovered. On the 10th day after laser irradiation, a large number of neovascularization appeared in the superficial layer of cortex, but the deep vessels did not recover. These results indicate that the method established in this study can observe the changes of blood vessel in cerebral ischemic region continuously, which lays a foundation for further quantitative study on the dynamics of embolized blood vessels and peripheral capillaries during the recovery of cerebral ischemia.

Small Molecular Weight Aldose (d-Glucose) and Basic Amino Acids (l-Lysine, l-Arginine) Increase the Occurrence of PAHs in Grilled Pork Sausages.

(1) Background: Amino acids and carbohydrates are widely used as additives in the food industry. These compounds have been proven to be an influencing factor in the production of chemical carcinogenic compounds polycyclic aromatic hydrocarbons (PAHs). However, the effect of the properties of the amino acids and carbohydrates on the production of PAHs is still little known. (2) Methods: We added different (i) R groups (the R group represents an aldehyde group in a glucose molecule or a ketone group in a fructose molecule); (ii) molecular weight carbohydrates; (iii) polarities, and (iv) acid-base amino acids to pork sausages. The effects of the molecular properties of carbohydrates and amino acids on the formation of PAHs in grilled pork sausages were investigated. (3) Results: The results showed that a grilled sausage with aldehyde-based d-glucose was capable of producing more PAHs than a sausage with keto-based d-fructose. A higher PAH content was determined in the grilled pork sausage when the smaller molecular weight, d-glucose, was added compared with the sausage where the larger molecular weight, 4-(α-d-glucosido)-d-glucose and cellulose were added. The addition of basic amino acids (l-lysine, l-arginine) was capable of producing more PAHs compared with the addition of acidic amino acids (l-glutamic acid, l-aspartate). When amino acid containing a benzene ring was added, a smaller volume of PAHs was produced compared with the addition of other amino acids. (4) Conclusions: Our study suggests that systematic consideration of molecule properties is necessary when using food additives (amino acids and carbohydrates) for food processing.

[Effect of heche chongcao capsule on expression of TNF-α protein in rats with chronic obstructive pulmonary disease].

OBJECTIVE: To observe the expression of tumor necrosis factor-alpha (TNF-α) protein in the lung tissue of rats with chronic obstructive pulmonary disease (COPD), and to evaluate the intervention and mechanism of Heche Chongcao Capsule (HCC). METHODS: The COPD rat model was prepared by exposure to cigarettes smoke plus intratracheal injecting lipopolysaccharide (LPS). Forty successfully modeled SD rats were randomly divided into the COPD model group, the control group, the low dose HCC group, the medium dose HCC group, and the high dose HCC group, 8 in each group. Meanwhile, a normal control group consisting of 6 rats was also set up. HCC at 0.25, 0.5, and 1.0 g/kg was administered to rats in the 3 dose HCC groups respectively by gastrogavage combined with Theophylline Sustained Release Tablet (TSRT). Rats in the control group were administered with TSRT by gastrogavage at 4.5 mg/kg, 1 mL/100 g each time, once daily. All medication lasted for 4 successive weeks. Equal volume of distilled water was administered by gastrogavage to rats in the COPD model group and the normal control group. Morphological changes of the lung tissue were observed under microscope. The expression of TNF-α protein in the lung tissue were also detected using Real-time PCR. RESULTS: Under electron microscope, the cilium in the tracheal epithelium was disorderly arranged, type I and II alveolar cells were degenerated, endoplasmic reticulum and mitochondria were swollen, the lamellar body was emptied, and free fragment could be seen inside alveolar space. Compared with the model group, all lesions were somewhat ameliorated in all medicated groups, especially in the medium dose HCC group. Compared with the model group, the expression of TNF-α protein decreased in all medicated groups, especially in the medium and low dose HCC groups (P < 0.05, P < 0.01). Compared with the control group, the expression of TNF-α protein decreased in the medium and low dose HCC groups (P < 0.05). CONCLUSION: HCC could effectively regulate the expression of TNF-α protein and inhibit airway inflammation reaction in COPD rats.

Insights into the in vitro digestibility of pork myofibrillar protein with different ionic polysaccharides from the perspective of gel characteristics.

In this work, the simulated gastrointestinal digestion of myofibrillar protein gels (MPGs) with anionic xanthan (XMP) and sodium alginate (SMP)/cationic chitosan (CSMP)/neutral curdlan (CMP) and konjac (KMP) was investigated to develop muscle-gelled foods with good qualities before and after eating. The results indicated that the neutral CMP and KMP groups had higher gel strength and protein digestibility than the CSMP group. Xanthan and sodium alginate facilitated myosin degradation in gastrointestinal digestion because of the weak wraps between protein and anionic polysaccharides, gaining plentiful peptides (1790 and 1692 respectively) with molecular weights below 2000 Da. Chitosan and neutral curdlan could improve the strength of MP gel but inhibited proteolysis and resulted in low contents of released amino acids via the strong cross-linked network blocking trypsin contact. This work provides a theoretical basis for developing low-fat meat products with good qualities and digestion behaviors by simply controlling the ionic types of polysaccharides.

Surveillance of COVID-19 vaccine safety among elderly persons aged 65 years and older.

BACKGROUND: Monitoring safety outcomes following COVID-19 vaccination is critical for understanding vaccine safety especially when used in key populations such as elderly persons age 65 years and older who can benefit greatly from vaccination. We present new findings from a nationally representative early warning system that may expand the safety knowledge base to further public trust and inform decision making on vaccine safety by government agencies, healthcare providers, interested stakeholders, and the public. METHODS: We evaluated 14 outcomes of interest following COVID-19 vaccination using the US Centers for Medicare & Medicaid Services (CMS) data covering 30,712,101 elderly persons. The CMS data from December 11, 2020 through Jan 15, 2022 included 17,411,342 COVID-19 vaccinees who received a total of 34,639,937 doses. We conducted weekly sequential testing and generated rate ratios (RR) of observed outcome rates compared to historical (or expected) rates prior to COVID-19 vaccination. FINDINGS: Four outcomes met the threshold for a statistical signal following BNT162b2 vaccination including pulmonary embolism (PE; RR = 1.54), acute myocardial infarction (AMI; RR = 1.42), disseminated intravascular coagulation (DIC; RR = 1.91), and immune thrombocytopenia (ITP; RR = 1.44). After further evaluation, only the RR for PE still met the statistical threshold for a signal; however, the RRs for AMI, DIC, and ITP no longer did. No statistical signals were identified following vaccination with either the mRNA-1273 or Ad26 COV2.S vaccines. INTERPRETATION: This early warning system is the first to identify temporal associations for PE, AMI, DIC, and ITP following BNT162b2 vaccination in the elderly. Because an early warning system does not prove that the vaccines cause these outcomes, more robust epidemiologic studies with adjustment for confounding, including age and nursing home residency, are underway to further evaluate these signals. FDA strongly believes the potential benefits of COVID-19 vaccination outweigh the potential risks of COVID-19 infection.

Background rates of adverse events of special interest for COVID-19 vaccine safety monitoring in the United States, 2019-2020.

BACKGROUND: The U.S. Food and Drug Administration (FDA) Biologics Effectiveness and Safety (BEST) Initiative conducts active surveillance of adverse events of special interest (AESI) after COVID-19 vaccination. Historical incidence rates (IRs) of AESI are comparators to evaluate safety. METHODS: We estimated IRs of 17 AESI in six administrative claims databases from January 1, 2019, to December 11, 2020: Medicare claims for adults ≥ 65 years and commercial claims (Blue Health Intelligence®, CVS Health, HealthCore Integrated Research Database, IBM® MarketScan® Commercial Database, Optum pre-adjudicated claims) for adults < 65 years. IRs were estimated by sex, age, race/ethnicity (Medicare), and nursing home residency (Medicare) in 2019 and for specific periods in 2020. RESULTS: The study included >100 million enrollees annually. In 2019, rates of most AESI increased with age. However, compared with commercially insured adults, Medicare enrollees had lower IRs of anaphylaxis (11 vs 12-19 per 100,000 person-years), appendicitis (80 vs 117-155), and narcolepsy (38 vs 41-53). Rates were higher in males than females for most AESI across databases and varied by race/ethnicity and nursing home status (Medicare). Acute myocardial infarction (Medicare) and anaphylaxis (all databases) IRs varied by season. IRs of most AESI were lower during March-May 2020 compared with March-May 2019 but returned to pre-pandemic levels after May 2020. However, rates of Bell's palsy, Guillain-Barré syndrome, narcolepsy, and hemorrhagic/non-hemorrhagic stroke remained lower in multiple databases after May 2020, whereas some AESI (e.g., disseminated intravascular coagulation) exhibited higher rates after May 2020 compared with 2019. CONCLUSION: AESI background rates varied by database and demographics and fluctuated in March-December 2020, but most returned to pre-pandemic levels after May 2020. It is critical to standardize demographics and consider seasonal and other trends when comparing historical rates with post-vaccination AESI rates in the same database to evaluate COVID-19 vaccine safety.

Near real-time surveillance of safety outcomes in US COVID-19 vaccine recipients aged 12 to 64 years.

BACKGROUND: Active monitoring of safety outcomes following COVID-19 vaccination is critical to understand vaccine safety and can provide early detection of rare outcomes not identified in pre-licensure trials. We present findings from an early warning rapid surveillance system in three large commercial insurance databases including more than 16 million vaccinated individuals. METHODS: We evaluated 17 outcomes of interest following COVID-19 vaccination among individuals aged 12-64 years in Optum, HealthCore, and CVS Health databases from December 11, 2020, through January 22, 2022, January 7, 2022, and December 31, 2021, respectively. We conducted biweekly or monthly sequential testing and generated rate ratios (RR) of observed outcome rates compared to historical (or expected) rates prior to COVID-19 vaccination. FINDINGS: Among 17 outcomes evaluated, 15 did not meet the threshold for statistical signal in any of the three databases. Myocarditis/pericarditis met the statistical threshold for a signal following BNT162b2 in two of three databases (RRs: 1.83-2.47). Anaphylaxis met the statistical threshold for a signal in all three databases following BNT162b2 vaccination (RRs: 4.48-10.86) and mRNA-1273 vaccination (RRs: 7.64-12.40). DISCUSSION: Consistent with published literature, our near-real time monitoring of 17 adverse outcomes following COVID-19 vaccinations identified signals for myocarditis/pericarditis and anaphylaxis following mRNA COVID-19 vaccinations. The method is intended for early detection of safety signals, and results do not imply a causal effect. Results of this study should be interpreted in the context of the method's utility and limitations, and the validity of detected signals must be evaluated in fully adjusted epidemiologic studies.

Multiplex-Heterogeneous Network-Based Capturing Potential SNP "Switches" of Pathways Associating With Diverse Disease Characteristics of Asthma.

Asthma is a complex heterogeneous respiratory disorder. In recent years nubbly regions of the role of genetic variants and transcriptome including mRNAs, microRNAs, and long non-coding RNAs in the pathogenesis of asthma have been separately excavated and reported. However, how to systematically integrate and decode this scattered information remains unclear. Further exploration would improve understanding of the internal communication of asthma. To excavate new insights into the pathogenesis of asthma, we ascertained three asthma characteristics according to reviews, airway inflammation, airway hyperresponsiveness, and airway remodeling. We manually created a contemporary catalog of corresponding risk transcriptome, including mRNAs, miRNAs, and lncRNAs. MIMP is a multiplex-heterogeneous networks-based approach, measuring the relevance of disease characteristics to the pathway by examining the similarity between the determined vectors of risk transcriptome and pathways in the same low-dimensional vector space. It was developed to enable a more concentrated and in-depth exploration of potential pathways. We integrated experimentally validated competing endogenous RNA regulatory information and the SNPs with significant pathways into the ceRNA-mediated SNP switching pathway network (CSSPN) to analyze ceRNA regulation of pathways and the role of SNP in these dysfunctions. We discovered 11 crucial ceRNA regulations concerning asthma disease feature pathway and propose a potential mechanism of ceRNA regulatory SNP → gene → pathway → disease feature effecting asthma pathogenesis, especially for MALAT1 (rs765499057/rs764699354/rs189435941) → hsa-miR-155 → IL13 (rs201185816/rs1000978586/rs202101165) → Interleukin-4 and Interleukin-13 signaling → inflammation/airway remodeling and MALAT1 (rs765499057/rs764699354/rs189435941) → hsa-miR-155 → IL17RB (rs948046241) → Interleukin-17 signaling (airway remodeling)/Cytokine-cytokine receptor interaction (inflammation). This study showed a systematic and propagable workflow for capturing the potential SNP "switch" of asthma through text and database mining and provides further information on the pathogenesis of asthma.

[Effect of 90 percentage portal branch ligation on liver regeneration and expression of metalloproteinase and tissue inhibitors of metalloproteinases in rats].

OBJECTIVE: To investigate the effect of 90% portal branch ligation on liver regeneration and expression of metalloproteinases and tissue inhibitors of metalloproteinases in rats. METHODS: Ninety-six SD rats were randomly divided into Sham-PBL group and portal vein branches ligation group. The weight of both ligated and unligated lobes of liver were measured at post operation day (POD) 0.5, 1, 3, 5, 7, 14, 21 and 28. The morphological changes of the non-ligated liver lobes were observed by microscope. The expression of PCNA, MMP2, MMP9 and TIMP2 of the non-ligated liver lobes were studied by immunohistochemistry. RESULTS: 1) 95.8% rats survived from the ligation of 90% portal branch. Hepatic lobe at the ligated side diminished progressively after ligation, whereas the lobes of the unligated side underwent compensatory regeneration. The ratio of non-ligated lobes weight to the whole liver increased slowly within 1d, speeded up significantly during 1-5d period, increased slowly after POD5, and got the plateau stag at POD7; 2) PCNA index were markedly increased within POD 0.5-3 (P < 0.01). It reached the peak at POD5 and decreased slightly at POD7, but still higher than Sham-PBL group level, then gradually returned to normal. 3) The expression of MMP2,MMP9 and TIMP2 in the non-ligated liver lobes were markedly increased at 1d. It reached the peak at POD7 and gradually returned to normal within POD7-28. 4) The MMP2 and PCNA in liver had a positive correlation at POD 0.5, 1, 5, 7, 14. The expressions of MMP9 and PCNA had a positive correlation at POD 0.5, 1, 7, 21. CONCLUSION: The expressions of TIMP2 and PCNA had a positive correlation at POD1, 7, 14, 21. The expression of MMP2, MMP9 and TIMP2 of the non-ligated liver lobes is markedly increased at POD1. It reaches the peak at POD7, and dropped to normal level gradually. The expressions of MMP2, MMP9 and TIMP2 and PCNA were correlated in 90% portal branch Ligation rats. The expression of MMP2,MMP9 and TIMP2 may play a pivotal role in liver regeneration.

Gastrointestinal digestion and cecal fermentation of a mixed gel of lean pork meat and resistant starch in mice.

The sensitivity of meat gel to digestive enzymes and the overall digestion pattern of the meat product is vital, and exerts an important influence on the growth and metabolism of mice. In order to provide a comprehensive understanding for better usage of resistant starch (RS) in functional meat products, the effects of a mixed gel (MS, a cooked mixture) of lean pork meat and RS on the gastrointestinal digestion and cecal fermentation of mice were investigated via comparing with those of RS-free meat gel (M) and the addition of RS to meat gel (M + S). The results showed that both M + S and MS promoted gastrointestinal digestion and cecal fermentation in mice. Specifically, the MS diet contributed to the hydrolysis of proteins, the formation of beneficial amino acids, and cecal health in spite of the larger particle size for digestion than that of the M + S group. Collectively, mixed gels of meat and RS are prospective for developing healthier meat products.

Effect of salt mixture on flavor of reduced-sodium restructured bacon with ultrasound treatment.

Flavor loss from sodium reduction is a large challenge faced in the meat industry. The effects of salt mixture (KCl: CaCl2 = 1:1, w/w) content (0%-1.0%) on flavor of reduced-sodium (1.5% NaCl) restructured bacon with ultrasound treatment (UT, 600 w for 30 min) were investigated. The results showed that 0.5% salt mixture (0.25% KCl and 0.25% CaCl2) could significantly (p < .05) enhance the lipid oxidation, the protein oxidation, and the formation of free amino acids of reduced-sodium UT-restructured bacon and could also markedly (p < .05) improve its flavor and the overall quality of sensory evaluation via promoting the release of five kinds of volatile phenolic compounds (o-cresol, m-cresol, 2-methoxy-phenol, 2-methoxy-4-methylphenol, and 2-methoxy-5-methylphenol) and the formation of five kinds of volatile aldehyde compounds (hexanal, nonanal, decanal, furfural, and 5-methyl furfural). It is interesting to understand the mechanism for the effect of salt mixture on flavor and to efficiently develop a technique for improving the flavor of reduced-sodium products in the meat industry.

Induction of germline apoptosis by cobalt and relevant signal transduction pathways in Caenorhabditis elegans.

Many investigations showed that cobalt exposure could induce apoptosis both in cells and tissues. However, appropriate in vivo animal models to assess the underlying mechanisms of cobalt-induced apoptosis are currently unavailable. The model organism, Caenorhabditis elegans, has been shown to be a good model for evaluating many biological processes. This study detected significant cobalt induced germline cell apoptosis after 12-h exposure; thus demonstrating that C. elegans could be a mammalian in vivo substitute model to study mechanisms of apoptosis. Then knockout gene C. elegans strains were utilized to investigate the relationship between cobalt-induced apoptosis and relevant signal pathways, which were involved in DNA damage and repair, apoptosis regulation, and damage signal transduction. The results presented here demonstrated that cobalt-induced apoptosis was independent of the DNA damage response gene, such as hus-1, p53/cep-1, and egl-1. The loss-of-function of the genes that related to JNK and p38 MAPK signaling cascades suppressed cobalt-induced germline apoptosis, while ERK signaling cascades have no effect on the cobalt-induced germline apoptosis.

Combined effect of CaCl2 and high pressure processing on the solubility of chicken breast myofibrillar proteins under sodium-reduced conditions.

The combined effect of calcium chloride (CaCl2) (20-100 mM) and high pressure processing (HPP, 200 MPa) on the solubility of myofibrillar proteins (MP) was investigated under sodium-reduced conditions (0.3 M sodium chloride). The results revealed that HPP combined with low concentrations (<40 mM) of CaCl2 synergistically increased the solubility of MP, but an antagonistic effect occurred when a high concentration (100 mM) of CaCl2 was present. This synergistic effect could be attributed to a mildly destabilized conformation of myosin, an increased surface hydrophobicity and the decreased total sulfhydryl group content of MP. However, CaCl2 at 100 mM destabilized myosin to a larger extent and induced non-disulfide covalent cross-linking of S-1 subfragment in pressurized MP, thus attenuating the solubilizing effect of HPP on the myosin heavy chain, resulting in the antagonistic effect. Therefore, HPP in combination with low-level CaCl2 (<40 mM) may improve the functional properties of sodium-reduced meat products.

[Effect of particulate matter 2.5 at urban centre of Hangzhou on lung impairment in rats].

OBJECTIVE: To explore the effect of ambient particle matter 2.5 (PM2.5) collected in the urban center of Hangzhou on the lung injury of rats and on the activating of endoplasmic reticulum pathway. METHODS: PM2.5 samples were collected on quartz fiber filters using a PM2.5 high-volume air sampler in the urban area of Hangzhou. The collected PM2.5 particles were extracted in ultrapure water and concentrated by vacuum freeze-drying. Twenty-four male Sprague-Dawly (SD) rats were randomly divided into 3 groups:saline control group, low dose PM2.5 exposure group (5 mg/kg BW) and high dose PM2.5 exposure groups (25 mg/kg BW). Each group received intratracheal instillation of PM2.5, once a week for 4 weeks. Twenty-four hours after the last exposure, the rats were narcotized and sacrificed, left lung was isolated and fixed with 4% paraformaldehyde for histopathological detection. The bronchoalveolar lavage fluid (BALF) was collected from the right lung. The total antioxidant capacity (T-AOC) level, the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) in BALF were detected by chemical colorimetry. The level of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß) and interleukin-6 (IL-6) cytokines in BALF was measured by enzyme linked immunosorbent assay (ELISA). And the protein expressions of glucose-regulated protein 78 (GRP78), phosphorylated protein kinase receptor-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic translation initiation factor (p-eIF2α), transcription factors C/EBP homologue protein (CHOP), inositol-requiring enzyme 1α (IRE1α) and X-box binding protein 1 (XBP1) in lung tissue were determined by Western blotting. RESULTS: Compared with control group, rats in both low dose (5 mg/kg) and high dose (25 mg/kg) PM2.5-treated groups showed obviously dose-dependent pulmonary toxicity including thickening of alveolar walls, narrowing of alveolar space, interstitial hyperplasia and inflammatory cell infiltration. Compared with control group, T-AOC level and the SOD activity in BALF in both PM2.5-treated groups were decreased dose-dependently (P<0.05), whereas the LDH activity in BALF were increased in a dose-dependent manner (P<0.05). Exposure to PM2.5 resulted in a increasing of the release of proinflammatory cytokines including TNF-α, IL-1ß and IL-6 in rat lung in a dose-dependent manner (P<0.05). The levels of GRP78, p-PERK, p-eIF2α, CHOP, IRE1α and spliced XBP1 (XBP1-S) were significantly up-regulated, whereas the level of unspliced XBP1 (XBP1-U) was down-regulated in the rat lung tissue of high-dose PM2.5 treated group. CONCLUSIONS: The PM2.5 in the urban area of Hangzhou can significantly cause lung inflammatory injury in rats. Both oxidative stress and activation of ER stress pathways may be related to such PM2.5 inhalation-induced lung inflammatory injury.

[Inhibition of hepatitis B virus S gene and C gene expression by different 10-23 DNAzymes substrate-recognition domains].

OBJECTIVE: To explore the inhibition effects of 10-23 DNAzymes with different substrate-recognition domains targeting hepatitis B virus (HBV) S gene and C gene expression in 2.2.15 cells. METHODS: 10-23 DNAzymes with different substrate-recognition domains specific to HBV S gene open reading frame (ORF) A(157)UG and HBV C gene ORF A(1816)UG were designed and synthesized, respectively. Different 10-23 DNAzymes were transfected into 2.2.15 cells which is a stable HBV producing cell line. HBsAg and HBeAg secreted into culture media were detected by radioimmunoassay (RIA) and HBV DNA levels were measured by real-time PCR. 3-(4, 5-dimethylthiagol-2-yl)-2, 5-drphnyl tetrazolium bromide (MTT) assays were performed to evaluate cytotoxicity. RESULTS: HBsAg and HBeAg expressions were reduced by various DNAzymes (0.1 - 2.5 micromol/L) with different substrate-recognition domains after transfection. The antiviral effects of DNAzymes were apparent until 72 h post-transfection. The inhibition rates of the DNAzymes at the same dose on HBsAg and HBeAg in the same period of post-transfection were as the following: DrzBS-9 > DrzBS-8 > DrzBS-7; DrzBC-9 > DrzBC-8 > DrzBC-7. Among all the DNAzymes used, DrzBS-9 targeting S gene and DrzBC-9 targeting C gene were most potent, with HBsAg and HBeAg reduced 95% and 92% 48 h post-transfection at the dose of 2.5 micromol/L, respectively. The inhibition effects on HBV DNA by various DNAzymes with different substrate-recognition domains were of no significance. There were no evident cytotoxic effects of these DNAzymes in the range from 0.1 to 2.5 micromol/L. CONCLUSION: 10-23 DNAzymes with different substrate-recognition domains targeting HBV S gene and C gene mRNA possessed specific inhibition effects in 2.2.15 cells, and DrzBS-9 targeting S gene and DrzBC-9 targeting C gene were most potent.

[Cleavage of in vitro transcripts of hepatitis B virus C gene by 10-23 DNA enzyme].

OBJECTIVE: To investigate the cleavage activities of 10-23 DNA enzymes targeting at HBV C gene mRNA in vitro. METHODS: 10-23 DNA enzymes named DrzBC-7, DrzBC-8 and DrzBC-9 specific to HBV C gene ORF A1816UG were designed and synthesized. HBV C gene mRNA was obtained by in vitro transcription method. Cleavage activities were observed in vitro. The influence of MgCl2 concentration on RNA cleaving activity was examined with DrzBC-9. Values of kinetic parameters including Km, Kcat and Kcat/Km were calculated accordingly. RESULT: Targeted substrate mRNA with the size of 300 nt was obtained by transcription in vitro. Under the certain cleavage conditions, DrzBC-7, DrzBC-8 and DrzBC-9 all efficiently cleaved target mRNA at specific sites in vitro. Cleavage products of 109 nt and 191 nt were obtained. No cleavage occurred without MgCl2. The most efficient cleavage was obtained at 150 mmol x L(-1) MgCl2. The efficiency of cleavage did not increase when the MgCl2 concentration was more than 200 mmol x L(-1). The kinetic parameters, Km, Kcat and Kcat/Km for DrzBC-9 were 1.4x10(-9) mol x L(-1), 1.6 min(-1) and 1.1x10(9) mol x L(-1) x min(-1), respectively. CONCLUSION: 10-23 DNA enzymes targeting at HBV C gene mRNA possess the specific cleavage activities in vitro.

Role of original and modified Frey's procedures in chronic pancreatitis.

AIM: To retrospectively review patients with chronic pancreatitis (CP) treated with Frey's procedures between January 2009 and January 2014. METHODS: A retrospective review was performed of patients with CP treated with Frey's procedures between January 2009 and January 2014 in the Department of Pancreatic Surgery. A cross-sectional study of postoperative pain relief, quality of life (QoL), and alcohol and nicotine abuse was performed by clinical interview, letters and telephone interview in January 2016. QoL of patients was evaluated with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) version 3.0. The patients were requested to fill in the questionnaires by themselves via correspondence or clinical interview. RESULTS: A total of 80 patients were enrolled for analysis, including 44 who underwent the original Frey's procedure and 36 who underwent a modified Frey's procedure. The mean age was 46 years in the original group and 48 years in the modified group. Thirty-five male patients (80%) were in the original group and 33 (92%) in the modified group. There were no differences in the operating time, blood loss, and postoperative morbidity and mortality between the two groups. The mean follow-up was 50.3 mo in the original group and 48.7 mo in the modified group. There were no differences in endocrine and exocrine function preservation between the two groups. The original Frey's procedure resulted in significantly better pain relief, as shown by 5-year follow-up (P = 0.032), better emotional status (P = 0.047) and fewer fatigue symptoms (P = 0.028). When stratifying these patients by the M-ANNHEIM severity index, no impact was found on pain relief after the two types of surgery. CONCLUSION: The original Frey's procedure is as safe as the modified procedure, but the former yields better pain relief. The severity of CP does not affect postoperative pain relief.

Effect of Modified Wheat Gluten on Boiling Resistance Capacity of Pork Meatballs.

The effect of the modified wheat gluten (MWG) extender, prepared by alcalase-based hydrolysis and transglutaminase cross-linking, on meatballs was analyzed in this study. Here, we studied the effect of MWG addition on the boiling resistance capacity of pork meatballs (MB-MWG) at high temperature (100 °C) and increasing cooking time; meatballs with added soy protein isolates (MB-SPI) and raw wheat gluten (MB-WG) were used as references. The cooking loss, water-holding capacity (WHC), and textural properties of meatballs were investigated. The results revealed that MB-MWG showed lower cooking loss, which decreased by 49.16% compared to meatballs without added extenders when treated for 30 min. The WHC of MB-MWG significantly increased from 80.68% to 95.42%. The hardness, springiness, and chewiness (textural properties) of MB-MWG were also significantly increased by 97.05%, 6.68%, and 121.96%, respectively. The addition of MWG increased the cross-linking in meatballs during the cooking process, as indicated by the higher G'. SDS-PAGE indicated an obvious decrease in myosin heavy chain in MB-MWG cooked for 30 min at 100 °C, which was attributed to the interaction of myofibrillar proteins in pork meat with MWG. The nuclear magnetic resonance T2 relaxation time patterns indicated that MWG addition caused an increase in the bound water content, and decrease in the free water content, of meatballs. An analysis of the microstructures revealed that the MB-MWG formed the most regular and compact network. Therefore, MWG could be used as an ingredient to facilitate the processing of meat products.