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1.  Tea and human health: biomedical functions of tea active components and current issues*  
Originating in China, tea and tea planting have spread throughout the world since the middle of the Tang dynasty. Now people from 160 countries in the world are accustomed to tea drinking. A brief history of tea’s medicinal role in China and its spread to the world are introduced. The effectiveness of tea active components and tea drinking on major human diseases, including cancer, metabolic syndrome, cardiovascular disease, and neurodegenerative diseases, is discussed. Also presented are some related issues, such as the bioavailability of tea active components, the new formulations of tea polyphenols, and the safety for consumers of dietary supplements containing tea polyphenols.
PMCID: PMC4322420  PMID: 25644464
Tea; Cancer; Metabolic syndrome; Cardiovascular disease; Neurodegenerative disease; Bioavailability
2.  Cloning of a caffeoyl-coenzyme A O-methyltransferase from Camellia sinensis and analysis of its catalytic activity*  
Epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3"Me) present in leaves of Camellia sinensis has many beneficial biological activities for human health. However, EGCG3"Me occurs naturally in tea leaves in extremely limited quantities. Finding an enzyme from C. sinensis to catalyze the synthesis of EGCG3"Me is an alternative method to make up for the scarcity of EGCG3"Me in natural situations. In the present study, a complementary DNA (cDNA) encoding region and genomic DNA of the caffeoyl-coenzyme A O-methyltransferase (CCoAOMT) gene were isolated from C. sinensis (designated CsCCoAOMT). Nucleotide sequence analysis of CsCCoAOMT revealed an open reading frame of 738 bp that encodes a polypeptide with a predicted molecular weight of 28 kDa, which correlated well with the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The full-length DNA sequence (2678 bp) contained five exons and four introns. The deduced amino acid sequence of CsCCoAOMT shared 92% identity with CCoAOMTs from Codonopsis lanceolata and Betula luminifera. The catalytic activity of CsCCoAOMT was analyzed. Three monomethylated epigallocatechin-3-O-gallate (EGCG) compounds (EGCG4"Me, EGCG3"Me, and EGCG3'Me) were produced by CsCCoAOMT with K m in the micromolar range. Real-time polymerase chain reaction (RT-PCR) experiments indicated that the CsCCoAOMT transcript was present at low levels during the early stages of leaf maturity (the first leaf and bud on a shoot) but the relative expression was augmented at advanced stages of leaf maturity (the third or fourth leaf on a shoot), which accorded well with changes in EGCG3"Me content in fresh leaves. Hence, we concluded that CsCCoAOMT catalyzes the syntheses of methylated EGCGs.
PMCID: PMC4322421  PMID: 25644465
Tea (Camellia sinensis); O-methyltransferase; CsCCoAOMT; Prokaryotic expression; Catalytic activity; Methylated epigallocatechin-3-O-gallate (EGCG)
3.  Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application*  
The cytochrome P450 gene CYP81A6 confers tolerance to bentazon and metsulfuron-methyl, two selective herbicides widely used for weed control in rice and wheat fields. Knockout mutants of CYP81A6 are highly susceptible to both herbicides. The present study aimed to characterize the CYP81A6 expression in rice. Quantitative real-time polymerase chain reaction (PCR) analyses demonstrated that foliar treatment of bentazon (500 mg/L) greatly induced expression of CYP81A6 in both wild-type (Jiazhe B) and its knockout mutant (Jiazhe mB): a 10-fold increase at 9 h before returning to basal levels at 24 h in Jiazhe B, while in the mutant the expression level rose to >20-fold at 12 h and maintained at such high level up to 24 h post exposure. In contrast, metsulfuron-methyl (500 mg/L) treatment did not affect the expression of CYP81A6 in Jiazhe B within 80 h; thereafter the expression peaked at 120 h and returned gradually to basal levels by Day 6. We suggest that a metabolite of metsulfuron-methyl, 1H-2,3-benzothiazin-4-(3H)-one-2,2-dioxide, is likely to be responsible for inducing CYP81A6 expression, rather than the metsulfuron-methyl itself. Use of a promoter-GUS reporter construct (CYP81A6Pro::GUS) demonstrated that CYP81A6 was constitutively expressed throughout the plant, with the highest expression in the upper surfaces of leaves. Subcellular localization studies in rice protoplasts showed that CYP81A6 was localized in the endoplasmic reticulum. These observations advance our understanding of CYP81A6 expression in rice, particularly its response to the two herbicides.
PMCID: PMC4322422  PMID: 25644466
CYP81A6; Bentazon; Metsulfuron-methyl; Expression induction; Xenobiotics
4.  Lead accumulation and tolerance of Moso bamboo (Phyllostachys pubescens) seedlings: applications of phytoremediation*  
A hydroponics experiment was aimed at identifying the lead (Pb) tolerance and phytoremediation potential of Moso bamboo (Phyllostachys pubescens) seedlings grown under different Pb treatments. Experimental results indicated that at the highest Pb concentration (400 μmol/L), the growth of bamboo seedlings was inhibited and Pb concentrations in leaves, stems, and roots reached the maximum of 148.8, 482.2, and 4282.8 mg/kg, respectively. Scanning electron microscopy revealed that the excessive Pb caused decreased stomatal opening, formation of abundant inclusions in roots, and just a few inclusions in stems. The ultrastructural analysis using transmission electron microscopy revealed that the addition of excessive Pb caused abnormally shaped chloroplasts, disappearance of endoplasmic reticulum, shrinkage of nucleus and nucleolus, and loss of thylakoid membranes. Although ultrastructural analysis revealed some internal damage, even the plants exposed to 400 μmol/L Pb survived and no visual Pb toxicity symptoms such as necrosis and chlorosis were observed in these plants. Even at the highest Pb treatment, no significant difference was observed for the dry weight of stem compared with controls. It is suggested that use of Moso bamboo as an experimental material provides a new perspective for remediation of heavy metal contaminated soil owing to its high metal tolerance and greater biomass.
PMCID: PMC4322423  PMID: 25644467
Moso bamboo; Pb; Phytoremediation; Scanning electron microscopy; Transmission electron microscopy
5.  Assessing winter oilseed rape freeze injury based on Chinese HJ remote sensing data*  
The winter oilseed rape (Brassica napus L.) accounts for about 90% of the total acreage of oilseed rape in China. However, it suffers the risk of freeze injury during the winter. In this study, we used Chinese HJ-1A/1B CCD sensors, which have a revisit frequency of 2 d as well as 30 m spatial resolution, to monitor the freeze injury of oilseed rape. Mahalanobis distance-derived growing regions in a normal year were taken as the benchmark, and a mask method was applied to obtain the growing regions in the 2010–2011 growing season. The normalized difference vegetation index (NDVI) was chosen as the indicator of the degree of damage. The amount of crop damage was determined from the difference in the NDVI before and after the freeze. There was spatial variability in the amount of crop damage, so we examined three factors that may affect the degree of freeze injury: terrain, soil moisture, and crop growth before the freeze. The results showed that all these factors were significantly correlated with freeze injury degree (P<0.01, two-tailed). The damage was generally more serious in low-lying and drought-prone areas; in addition, oilseed rape planted on south- and west-oriented facing slopes and those with luxuriant growth status tended to be more susceptible to freeze injury. Furthermore, land surface temperature (LST) of the coldest day, soil moisture, pre-freeze growth and altitude were in descending order of importance in determining the degree of damage. The findings proposed in this paper would be helpful in understanding the occurrence and severity distribution of oilseed rape freeze injury under certain natural or vegetation conditions, and thus help in mitigation of this kind of meteorological disaster in southern China.
PMCID: PMC4322424  PMID: 25644468
Brassica napus; Freeze injury; Remote sensing; Crop monitoring; HJ-CCD
6.  Treatment of turtle aquaculture effluent by an improved multi-soil-layer system*  
Concentrated turtle aquaculture effluent poses an environmental threat to water bodies, and therefore needs to be treated prior to disposal. This study was conducted to assess the effect of multi-soil-layer (MSL) systems treating turtle aquaculture effluent with adding different amounts of sludge. Four MSL systems were constructed with dry weight ratios of sludge with 0%, 5%, 10%, and 20% (MSL 1, MSL 2, MSL 3, and MSL 4, respectively). The turtle aquaculture effluent had an average chemical oxygen demand (COD), ammonia nitrogen (NH4 +-N) and total nitrogen (TN) concentration of 288.4, 213.4, and 252.0 mg/L, respectively. The COD/TN (C/N) ratio was 1.2. The results showed that the four MSL systems could effectively treat the COD, NH4 +-N, and TN, and MSL 4 showed significantly improved NH4 +-N removal efficiency, suggesting the potential of sludge addition to improve the turtle aquaculture effluent treatment. The average COD, TN, and NH4 +-N removal efficiencies of MSL 4 were 70.3%, 66.5%, and 72.7%, respectively. To further interpret the contribution of microorganisms to the removal, the microbial community compositions and diversities of the four MSL systems were measured. Comparisons of the denaturing gradient gel electrophoresis (DGGE) profiles revealed that the amount of nitrifying bacteria and diversity in MSL 4 were higher than those in the other three systems. We concluded that adding 20% of sludge improved the NH4 +-N removal and stability of the system for nitrification, due to the enrichment of the nitrifying bacteria in MSL 4.
PMCID: PMC4322425  PMID: 25644469
Turtle aquaculture effluent; Multi-soil-layer (MSL) system; Sludge; Microbial community diversity
7.  Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody*  
Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.
PMCID: PMC4322426  PMID: 25644470
Freshness; Royal jelly; Major royal jelly protein 1 (MRJP1); Enzyme-linked immunosorbent assay (ELISA); High specific antibody
9.  Epigenetic and metabolic regulation of breast cancer stem cells*  
Breast cancer has a relatively high mortality rate in women due to recurrence and metastasis. Increasing evidence has identified a rare population of cells with stem cell-like properties in breast cancer. These cells, termed cancer stem cells (CSCs), which have the capacity for self-renewal and differentiation, contribute significantly to tumor progression, recurrence, drug resistance and metastasis. Clarifying the mechanisms regulating breast CSCs has important implications for our understanding of breast cancer progression and therapeutics. A strong connection has been found between breast CSCs and epithelial mesenchymal transition (EMT). In addition, recent studies suggest that the maintenance of the breast CSC phenotype is associated with epigenetic and metabolic regulation. In this review, we focus on recent discoveries about the connection between EMT and CSC, and advances made in understanding the roles and mechanisms of epigenetic and metabolic reprogramming in controlling breast CSC properties.
PMCID: PMC4288940  PMID: 25559951
Cancer stem cells (CSCs); Epithelial mesenchymal transition (EMT); Epigenetic modification; Metabolic reprogramming; Breast cancer
10.  MicroRNAs in breast cancer: oncogene and tumor suppressors with clinical potential*  
MicroRNAs (miRs) are small single-stranded RNA molecules, which function as key negative regulators of post-transcriptional modulation in almost all biological processes. Abnormal expression of microRNAs has been observed in various types of cancer including breast cancer. Great efforts have been made to identify an association between microRNA expression profiles and breast cancer, and to understand the functional role and molecular mechanism of aberrant-expressed microRNAs. As research progressed, ‘oncogenic microRNAs’ and ‘tumor suppressive microRNAs’ became a focus of interest. The potential of candidate microRNAs from both intercellular (tissue) and extracellular (serum) sources for clinical diagnosis and prognosis was revealed, and treatments involving microRNA achieved some amazing curative effects in cancer disease models. In this review, advances from the most recent studies of microRNAs in one of the most common cancers, breast cancer, are highlighted, especially the functions of specifically selected microRNAs. We also assess the potential value of these microRNAs as diagnostic and prognostic markers, and discuss the possible development of microRNA-based therapies.
PMCID: PMC4288941  PMID: 25559952
Breast cancer; MicroRNA; Oncogene; Tumor suppressors; Diagnosis marker; MicroRNA-based therapy
11.  Hypoxia-inducible factor 1 and breast cancer metastasis*  
Accumulating evidence has shown that the hypoxic microenvironment, which is critical during cancer development, plays a key role in regulating breast cancer progression and metastasis. The effects of hypoxia-inducible factor 1 (HIF-1), a master regulator of the hypoxic response, have been extensively studied during these processes. In this review, we focus on the roles of HIF-1 in regulating breast cancer cell metastasis, specifically its effects on multiple key steps of metastasis, such as epithelial-mesenchymal transition (EMT), invasion, extravasation, and metastatic niche formation. We also discuss the roles of HIF-1-regulated non-coding RNAs in breast cancer metastasis, and therapeutic opportunities for breast cancer through targeting the HIF-1 pathway.
PMCID: PMC4288942  PMID: 25559953
Breast cancer; Hypoxia-inducible factor 1 (HIF-1); Metastasis
13.  Growth effects of botulinum toxin type A injected unilaterally into the masseter muscle of developing rats*  
Objective: To evaluate the effects of botulinum toxin type A (BTX-A) on mandible skeletal development by inducing muscle hypofunction. Methods: Four-week-old Sprague-Dawley rats (n=60) were divided into three groups: Group 1 animals served as controls and were injected with saline; Group 2 animals were injected unilaterally with BTX-A (the contralateral side was injected with saline); and Group 3 animals were injected bilaterally with BTX-A. In Group 2, the saline-injected side was designated the control side (Group 2-1), whereas the BTX-A-injected side was designated the experimental side (Group 2-2). After four weeks, the animals were sacrificed, dry skulls were prepared, and mandibles were measured. Results: In the unilateral group, the experimental side (Group 2-2) had reduced dimensions for all mandible measurements compared with the control side (Group 2-1), suggesting a local effect of BTX-A on mandible growth, likely due to muscle reduction. Conclusions: Localized BTX-A injection induced a change in craniofacial growth, and the skeletal effect was unilateral despite both sides of the mandible functioning as one unit.
PMCID: PMC4288944  PMID: 25559955
Botulinum toxin; Craniofacial growth; Mandible
14.  Effects of rapamycin against paraquat-induced pulmonary fibrosis in mice*  
Background and aims: Ingestion of paraquat (PQ), a widely used herbicide, can cause severe toxicity in humans, leading to a poor survival rate and prognosis. One of the main causes of death by PQ is PQ-induced pulmonary fibrosis, for which there are no effective therapies. The aim of this study was to evaluate the effects of rapamycin (RAPA) on inhibiting PQ-induced pulmonary fibrosis in mice and to explore its possible mechanisms. Methods: Male C57BL/6J mice were exposed to either saline (control group) or PQ (10 mg/kg body weight, intraperitoneally; test group). The test group was divided into four subgroups: a PQ group (PQ-exposed, non-treated), a PQ+RAPA group (PQ-exposed, treated with RAPA at 1 mg/kg intragastrically), a PQ+MP group (PQ-exposed, treated with methylprednisolone (MP) at 30 mg/kg intraperitoneally), and a PQ+MP+RAPA group (PQ-exposed, treated with MP at 30 mg/kg intraperitoneally and with RAPA at 1 mg/kg intragastrically). The survival rate and body weight of all the mice were recorded every day. Three mice in each group were sacrificed at 14 d and the rest at 28 d after intoxication. Lung tissues were excised and stained with hematoxylin-eosin (H&E) and Masson’s trichrome stain for histopathological analysis. The hydroxyproline (HYP) content in lung tissues was detected using an enzyme-linked immunosorbent assay (ELISA) kit. The expression of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) in lung tissues was detected by immunohistochemical staining and Western blotting. Results: A mice model of PQ-induced pulmonary fibrosis was established. Histological examination of lung tissues showed that RAPA treatment moderated the pathological changes of pulmonary fibrosis, including alveolar collapse and interstitial collagen deposition. HYP content in lung tissues increased soon after PQ intoxication but had decreased significantly by the 28th day after RAPA treatment. Immunohistochemical staining and Western blotting showed that RAPA treatment significantly down-regulated the enhanced levels of TGF-β1 and α-SMA in lung tissues caused by PQ exposure. However, RAPA treatment alone could not significantly ameliorate the lower survival rate and weight loss of treated mice. MP treatment enhanced the survival rate, but had no significant effects on attenuating PQ-induced pulmonary fibrosis or reducing the expression of TGF-β1 and α-SMA. Conclusions: This study demonstrates that RAPA treatment effectively suppresses PQ-induced alveolar collapse and collagen deposition in lung tissues through reducing the expression of TGF-β1 and α-SMA. Thus, RAPA has potential value in the treatment of PQ-induced pulmonary fibrosis.
PMCID: PMC4288945  PMID: 25559956
Paraquat; Pulmonary fibrosis; Rapamycin; Transforming growth factor-β1; α-Smooth muscle actin; Methylprednisolone
15.  Chronic corticosterone exposure reduces hippocampal glycogen level and induces depression-like behavior in mice*  
Long-term exposure to stress or high glucocorticoid levels leads to depression-like behavior in rodents; however, the cause remains unknown. Increasing evidence shows that astrocytes, the most abundant cells in the central nervous system (CNS), are important to the nervous system. Astrocytes nourish and protect the neurons, and serve as glycogen repositories for the brain. The metabolic process of glycogen, which is closely linked to neuronal activity, can supply sufficient energy substrates for neurons. The research team probed into the effects of chronic corticosterone (CORT) exposure on the glycogen level of astrocytes in the hippocampal tissues of male C57BL/6N mice in this study. The results showed that chronic CORT injection reduced hippocampal neurofilament light protein (NF-L) and synaptophysin (SYP) levels, induced depression-like behavior in male mice, reduced hippocampal glycogen level and glycogen synthase activity, and increased glycogen phosphorylase activity. The results suggested that the reduction of the hippocampal glycogen level may be the mechanism by which chronic CORT treatment damages hippocampal neurons and induces depression-like behavior in male mice.
PMCID: PMC4288946  PMID: 25559957
Glycogen; Corticosterone; Stress; Depression; Hippocampus
16.  Epidemiology and microbiology of nosocomial bloodstream infections: analysis of 482 cases from a retrospective surveillance study*  
In many traditional Chinese medicine (TCM) hospitals, most patients are elderly with chronic diseases. Nosocomial bloodstream infections (nBSIs) are an important cause of morbidity and mortality. A retrospective surveillance study was performed to examine the epidemiology and microbiology of nBSIs in a TCM hospital from 2009 to 2011. A total of 482 patients with nBSIs were included in the study period. The incidence rate was 5.7/1000 admissions. Escherichia coli (25.5%) was the most common Gram-negative and coagulase-negative staphylococcus (CoNS) (14.1%) was the most common Gram-positive organism isolated. One-third of the E. coli and Klebsiella pneumoniae isolated from the nBSIs were the third-generation cephalosporin-resistant. Half of the Acinetobacter species isolates were resistant to imipenem. Of all the CoNS isolates, 90.7% were resistant to methicillin. Carbapenems and glycopeptide were the most frequently used for nBSI therapy. Only about one-third of patients (157/482) received appropriate empirical therapy. Septic shock, hemodialysis, Pitt bacteremia score >4, urinary tract infection, and appropriate empirical therapy were most strongly associated with 28-d mortality. The incidence of nBSIs was low in the TCM hospital but the proportion of nBSIs due to antibiotic-resistant organisms was high. A high Pitt bacteremia score was one of the most important risk factors for mortality in nBSIs. Therefore, the implementation of appropriate empirical therapy is crucial to improve the clinical outcome of nBSIs.
PMCID: PMC4288947  PMID: 25559958
Nosocomial bloodstream infection; Traditional Chinese medicine hospital; Epidemiology; Microbiology
17.  Elevated homocysteine levels and risk of cardiovascular and all-cause mortality: a meta-analysis of prospective studies 
Objective: To investigate whether elevated homocysteine levels were a predictor of subsequent coronary heart disease (CHD) mortality, cardiovascular mortality or all-cause mortality in the general population by a meta-analysis. Methods: In a systematic search conducted in the databases of PubMed and Embase prior to October 2013, we identified relevant prospective observational studies evaluating the association between baseline homocysteine levels and CHD mortality, cardiovascular or all-cause mortality in the general population. Pooled adjust risk ratio (RR) and corresponding 95% confidence interval (CI) were calculated separately for categorical risk estimates and continuous risk estimates. Results: Twelve studies with 23 623 subjects were included in the meta-analysis. Comparing the highest to lowest homocysteine level categories, CHD mortality increased by 66% (RR 1.66; 95% CI 1.12–2.47; P=0.012), cardiovascular mortality increased by 68% (RR 1.68; 95% CI 1.04–2.70; P=0.033), and all-cause mortality increased by 93% (RR 1.93; 95% CI 1.54–2.43; P<0.001). Moreover, for each 5 μmol/L homocysteine increment, the pooled RR was 1.52 (95% CI 1.26–1.84; P<0.001) for CHD mortality, 1.32 (95% CI 1.08–1.61; P=0.006) for cardiovascular mortality, and 1.27 (95% CI 1.03–1.55; P=0.023) for all-cause mortality. Conclusions: Elevated homocysteine levels are an independent predictor for subsequent cardiovascular mortality or all-cause mortality, and the risks were more pronounced among elderly persons.
PMCID: PMC4288948  PMID: 25559959
Homocysteine; Coronary heart disease; Cardiovascular mortality; All-cause mortality; Meta-analysis
18.  Zirconium oxide ceramic foam: a promising supporting biomaterial for massive production of glial cell line-derived neurotrophic factor*  
This study investigated the potential application of a zirconium oxide (ZrO2) ceramic foam culturing system to the production of glial cell line-derived neurotrophic factor (GDNF). Three sets of ZrO2 ceramic foams with different pore densities of 10, 20, and 30 pores per linear inch (PPI) were prepared to support a 3D culturing system. After primary astrocytes were cultured in these systems, production yields of GDNF were evaluated. The biomaterial biocompatibility, cell proliferation and activation of cellular signaling pathways in GDNF synthesis and secretion in the culturing systems were also assessed and compared with a conventional culturing system. In this study, we found that the ZrO2 ceramic foam culturing system was biocompatible, using which the GDNF yields were elevated and sustained by stimulated cell proliferation and activation of signaling pathways in astrocytes cultured in the system. In conclusion, the ZrO2 ceramic foam is promising for the development of a GDNF mass production device for Parkinson’s disease treatment.
PMCID: PMC4265555  PMID: 25471830
Zirconium oxide; Ceramic foam; Glial cell line-derived neurotrophic factor (GDNF); Parkinson’s disease
19.  Dynamic changes of serum cholinesterase activity after severe trauma*  
Objective: The aim of the present study was to examine dynamic changes in serum cholinesterase (ChE) activity during early-stage severe trauma and the clinical significance of these changes. Methods: This prospective, observational study included 81 patients with severe trauma who were treated between October 2011 and April 2013 in the emergency intensive care unit (EICU) of a university-affiliated, tertiary-care, grade A general hospital in China. Serum ChE activity was measured on Days 1, 3, and 7 post-injury. The correlation of dynamic changes in serum ChE activity with trauma severity and prognosis was assessed. Correlations between changes in serum ChE activity after injury and albumin (ALB), prealbumin (PAB), transferrin (TRF), and C-reactive protein (CRP) levels were also analyzed. Results: Serum ChE activity in trauma patients was 42.3%–50.2% lower on Days 1, 3, and 7 compared with the control (P<0.001 for all time points), and it continued to decrease after Day 7 in both the survival and death subgroups. In the subgroup with an injury severity score (ISS) of ≤25, serum ChE activity initially decreased, but eventually increased. However, activity decreased continuously in the ISS>25 subgroup. ChE activity was significantly lower in both the death and the ISS>25 subgroups than in the survival and ISS≤25 subgroups on Days 1, 3, and 7 after injury. Activity was negatively correlated with ISS and acute physiology and chronic health evaluation III (APACHE III) at all time points. When comparing the receiver operating characteristic (ROC) curves for predicting prognosis, the area under the curve (AUC) in the plot of serum ChE was similar to the AUCs in plots of ISS and APACHE III, but significantly smaller than the AUC in the plot of the trauma and injury severity score (TRISS). Serum ChE activity was positively correlated with ALB, PAB, and TRF at all time points post-injury. Activity was not significantly correlated with CRP on Day 1, but was significantly and negatively correlated with CRP on Days 3 and 7. Conclusions: There is a significant decrease in serum ChE activity after severe trauma. Serum ChE may be regarded as a negative acute phase protein (APP) and the dynamic changes in serum ChE may be useful as an auxiliary indicator for evaluating trauma severity and predicting prognosis.
PMCID: PMC4265556  PMID: 25471831
Multiple trauma; Cholinesterase (ChE); Acute phase protein
20.  USP11 regulates p53 stability by deubiquitinating p53*  
The p53 tumor suppressor protein coordinates the cellular responses to a broad range of cellular stresses, leading to DNA repair, cell cycle arrest or apoptosis. The stability of p53 is essential for its tumor suppressor function, which is tightly controlled by ubiquitin-dependent degradation primarily through its negative regulator murine double minute 2 (Mdm2). To better understand the regulation of p53, we tested the interaction between p53 and USP11 using co-immunoprecipitation. The results show that USP11, an ubiquitin-specific protease, forms specific complexes with p53 and stabilizes p53 by deubiquitinating it. Moreover, down-regulation of USP11 dramatically attenuated p53 induction in response to DNA damage stress. These findings reveal that USP11 is a novel regulator of p53, which is required for p53 activation in response to DNA damage.
PMCID: PMC4265557  PMID: 25471832
p53; USP11; Deubiquitination; Stability
21.  Therapeutic detoxification of quercetin against carbon tetrachloride-induced acute liver injury in mice and its mechanism*  
This study observes the therapeutic detoxification of quercetin, a well-known flavonoid, against carbon tetrachloride (CCl4) induced acute liver injury in vivo and explores its mechanism. Quercetin decreased CCl4-increased serum activities of alanine and aspartate aminotransferases (ALT/AST) when orally taken 30 min after CCl4 intoxication. The results of a histological evaluation further evidenced the ability of quercetin to protect against CCl4-induced liver injury. Quercetin decreased the CCl4-increased malondialdehyde (MDA) and reduced the glutathione (GSH) amounts in the liver. It also reduced the enhanced immunohistochemical staining of the 4-hydroxynonenal (4-HNE) in the liver induced by CCl4. Peroxiredoxin (Prx) 1, 2, 3, 5, 6, thioredoxin reductase 1 and 2 (TrxR1/2), thioredoxin 1 and 2 (Trx1/2), nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) all play critical roles in maintaining cellular redox homeostasis. Real-time polymerase chain reaction (PCR) results demonstrated that quercetin reversed the decreased mRNA expression of all those genes induced by CCl4. In conclusion, our results demonstrate that quercetin ameliorates CCl4-induced acute liver injury in vivo via alleviating oxidative stress injuries when orally taken after CCl4 intoxication. This protection may be caused by the elevation of the antioxidant capacity induced by quercetin.
PMCID: PMC4265558  PMID: 25471833
Hepatotoxicity; Oxidative stress; Peroxiredoxin (Prx); Nuclear factor erythroid 2-related factor 2 (Nrf2); TrxR; Trx; HO-1
22.  Predictive factors of cardiac rupture in patients with ST-elevation myocardial infarction*  
Cardiac rupture (CR) is a potentially fatal mechanical complication of ST-elevation myocardial infarction (STEMI). We aimed to determine the incidence and risk factors of CR in Chinese STEMI patients. A total of 9798 consecutive STEMI patients from four centers in China were retrospectively analyzed, among which 178 patients had CR. STEMI patients without CR were chosen as a control group. Clinical characteristics were compared between STEMI patients with CR and those without CR. The incidence of CR in STEMI patients was 1.82%, and the 30-d mortality was up to 61.2%. CR patients were significantly older, more female, and associated with a longer time from onset of pain to hospital admission than their non-CR counterparts (P<0.001). More patients with anterior myocardial infarction (82.1%) were found in the CR group, and CR patients had significantly higher heart rates than the control group ((91±19) bpm vs. (71±16) bpm; P<0.001). In multiple-adjusted models, the independent risk factors of CR were advanced age, female gender, anaemia, increased heart rate, anterior myocardial infarction, increased white blood cell (WBC) count, delayed admission, and renal dysfunction. The level of hemoglobin remained a significant determinant factor of CR (OR (95% CI): 0.82 (0.75–0.89); P<0.001) after adjusting for various potential confounding factors. Counts of WBC also remained a significant determinant of the CR (OR (95% CI): 1.08 (1.04–1.12); P<0.001). A number of variables were independently related to CR. This study indicated, for the first time, that both hemoglobin and WBC levels were independently correlated with occurrence of CR.
PMCID: PMC4265559  PMID: 25471834
ST-elevation myocardial infarction; Risk factor; Anaemia; Cardiac rupture
23.  Peripheral deep anterior lamellar keratoplasty using a cryopreserved donor cornea for Terrien’s marginal degeneration*  
Objective: To evaluate the clinical efficacy of peripheral deep anterior lamellar keratoplasty (DALK) using a cryopreserved donor cornea for Terrien’s marginal degeneration (TMD). Methods: Thirty-one eyes of 27 patients with TMD underwent peripheral DALK using cryopreserved donor corneas. According to the distance between the inner edge of the lesion and the limbus, a ring-shaped or D-shaped DALK was performed. All grafts were stored at −20 °C. Cryopreserved corneoscleral rims were prepared for ring-shaped grafts and cryopreserved whole eyeballs were prepared for D-shaped grafts. The general conditions, intraoperative performance, postoperative corneal reconstruction, astigmatism, best corrected visual acuity (BCVA), and various complications were analyzed. Results: Ring-shaped DALK was performed in 28 eyes and D-shaped DALK was performed in 3 eyes. Postoperative follow-up time was (28.4±24.8) months. There was evidence of inflammation before surgery in 12 eyes (38.7%) and intraoperative perforation occurred in 13 eyes (41.9%). The corneal structures of all eyes were reconstructed. Postoperative astigmatism and BCVA showed improvement (both P=0.00) except for cases that underwent D-shaped DALK. Ten eyes (32.2%) developed transient ocular hypertension and one eye (3.2%) developed secondary glaucoma. No primary disease recurrence or corneal allograft rejection was observed. Conclusions: Peripheral DALK for TMD using cryopreserved donor tissue is an effective technique that eliminates rejection and extends the use of donor eyes. Inflammatory history or intraoperative perforation has no adverse effect on graft recovery. However, D-shaped DALK did not achieve good visual outcomes.
PMCID: PMC4265560  PMID: 25471835
Terrien’s marginal degeneration (TMD); Deep anterior lamellar keratoplasty (DALK); Cryopreservation
24.  Contour changes in human alveolar bone following tooth extraction of the maxillary central incisor 
The purpose of this study was to apply cone-beam computed tomography (CBCT) to observe contour changes in human alveolar bone after tooth extraction of the maxillary central incisor and to provide original morphological evidence for aesthetic implant treatment in the maxillary anterior area. Forty patients were recruited into the study. Each patient had two CBCT scans (CBCT I and CBCT II), one taken before and one taken three months after tooth extraction of maxillary central incisor (test tooth T). A fixed anatomic reference point was used to orient the starting axial slice of the two scans. On three CBCT I axial slices, which represented the deep, middle, and shallow layers of the socket, labial and palatal alveolar bone widths of T were measured. The number of sagittal slices from the start point to the pulp centre of T was recorded. On three CBCT II axial slices, the pulp centres of extracted T were oriented according to the number of moved sagittal slices recorded in CBCT I. Labial and palatal alveolar bone widths at the oriented sites were measured. On the CBCT I axial slice which represented the middle layer of the socket, sagittal slices were reconstructed. Relevant distances of T on the sagittal slice were measured, as were the alveolar bone width and tooth length of the opposite central incisor. On the CBCT II axial slice, which represented the middle layer of the socket, relevant distances recorded in CBCT I were transferred on the sagittal slice. The height reduction of alveolar bone on labial and palatal sides was measured, as were the alveolar bone width and tooth length of the opposite central incisor at the oriented site. Intraobserver reliability assessed by intraclass correlation coefficients (ICCs) was high. Paired sample t-tests were performed. The alveolar bone width and tooth length of the opposite central incisor showed no statistical differences (P<0.05). The labial alveolar bone widths of T at the deep, middle, and shallow layers all showed statistical differences. However, no palatal alveolar bone widths showed any statistical differences. The width reduction of alveolar bone was 1.2, 1.6, and 2.7 mm at the deep, middle, and shallow layers, respectively. The height reduction of alveolar bone on labial and palatal sides of T both showed statistical differences, which was 1.9 and 1.1 mm, respectively.
PMCID: PMC4265561  PMID: 25471836
Cone-beam computed tomography; Alveolar bone contour; Maxillary central incisor
25.  Development of the science of mass casualty incident management: reflection on the medical response to the Wenchuan earthquake and Hangzhou bus fire*  
Objective: In this paper, we review the previous classic research paradigms of a mass casualty incident (MCI) systematically and reflect the medical response to the Wenchuan earthquake and Hangzhou bus fire, in order to outline and develop an improved research paradigm for MCI management. Methods: We searched PubMed, EMBASE, China Wanfang, and China Biology Medicine (CBM) databases for relevant studies. The following key words and medical subject headings were used: ‘mass casualty incident’, ‘MCI’, ‘research method’, ‘Wenchuan’, ‘earthquake’, ‘research paradigm’, ‘science of surge’, ‘surge’, ‘surge capacity’, and ‘vulnerability’. Searches were performed without year or language restriction. After searching the four literature databases using the above listed key words and medical subject headings, related articles containing research paradigms of MCI, 2008 Wenchuan earthquake, July 5 bus fire, and science of surge and vulnerability were independently included by two authors. Results: The current progresses on MCI management include new golden hour, damage control philosophy, chain of survival, and three links theory. In addition, there are three evaluation methods (medical severity index (MSI), potential injury creating event (PICE) classification, and disaster severity scale (DSS)), which can dynamically assess the MCI situations and decisions for MCI responses and can be made based on the results of such evaluations. However, the three methods only offer a retrospective evaluation of MCI and thus fail to develop a real-time assessment of MCI responses. Therefore, they cannot be used as practical guidance for decision-making during MCI. Although the theory of surge science has made great improvements, we found that a very important factor has been ignored—vulnerability, based on reflecting on the MCI response to the 2008 Wenchuan earthquake and July 5 bus fire in Hangzhou. Conclusions: This new paradigm breaks through the limitation of traditional research paradigms and will contribute to the development of a methodology for disaster research.
PMCID: PMC4265562  PMID: 25471837
Mass casualty incident; Surge; Vulnerability; Earthquake; Fire incident

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