Pigment Characterization of the Giant-Colony-Forming Haptophyte Phaeocystis globosa in the Beibu Gulf Reveals Blooms of Different Origins.

The giant-colony-forming haptophyte Phaeocystis globosa has caused several large-scale blooms in the Beibu Gulf since 2011, but the distribution and dynamics of the blooms remained largely unknown. In this study, colonies of P. globosa, as well as membrane-concentrated phytoplankton samples, were collected during eight cruises in the Beibu Gulf from September 2016 to August 2017. Pigments were analyzed by high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD). The pigment 19'-hexanoyloxyfucoxanthin (hex-fuco), generally considered a diagnostic pigment for Phaeocystis, was not detected in P. globosa colonies in the Beibu Gulf, whereas 19'-butanoyloxyfucoxanthin (but-fuco) was found in all colony samples. Moreover, but-fuco in membrane-concentrated phytoplankton samples exhibited a similar distribution pattern to that of P. globosa colonies, suggesting that but-fuco provided the diagnostic pigment for bloom-forming P. globosa in the Beibu Gulf. Based on the distribution of but-fuco in different water masses in the region prior to the formation of intensive blooms, it is suggested that P. globosa blooms in the Beibu Gulf could originate from two different sources. IMPORTANCE Phaeocystis globosa has formed intensive blooms in the South China Sea and even around the world, causing huge social economic losses and environmental damage. However, little is known about the formation mechanism and dynamics of P. globosa blooms. 19'-Hexanoyloxyfucoxanthin (hex-fuco) is often used as the pigment proxy to estimate Phaeocystis biomass, while this is challenged by the giant-colony-forming P. globosa in the Beibu Gulf, which contains only 19'-butanoyloxyfucoxanthin (but-fuco) but not hex-fuco. Using but-fuco as a diagnostic pigment, we traced two different origins of P. globosa blooms in the Beibu Gulf. This study clarifies the development process of P. globosa blooms in the Beibu Gulf, which provides a basis for the early monitoring and prevention of the blooms.

Ecological wetland paradigm drives water source improvement in the stream network of Yangtze River Delta.

Jiaxing created a precedent using bypass riparian marshes to purify micro-polluted water sources in China. Pond-wetland complex with constructed root channel technology becomes a paradigm which can be analogized as "human-body wetland model" based on bionics or biomimetics. Heterogeneous plant-bed/ditch system with highly active land/water ecotone interfaces, especially meandering boundaries, breeds many biochemical reactions "living areas". Optimization of hydraulic regulation promotes redox environment alternations and wetland treatment efficiency. Here we reported a series of upgrades and performances in Guanjinggang wetland after the Shijiuyang prototype. Morphological reform of plant-bed/ditch system played a vital role. Spatially root channel zone was main force of wetland purification, and temporally the treatment effect was higher in low-temperature seasons indicating non-temperature dependent mechanisms worked. Water pollution comprehensive index improved steadily from IV to III, and comprehensive pollution load was reduced by ca. 40%-60%. Comprehensive evaluation function value further showed the gradients purification effect of the upgraded wetland. Ecological wetlands ameliorated source water quality, and reduced drinking water treatment reagents, thereby bringing about economic benefits. Through wetlands operation, people can see how the micro-polluted surface water becomes clear and clean, so promoting a significant social benefit. As a viable component of urban green space, wetlands could beautify regional eco-environment, freshen the air, increase urban ecological taste, and enhance the eco-environmental protection publicity. Thus, the multifunctional service values and indirect benefits are substantial. Jiaxing ecological wetlands provide a typical paradigm for water pollution remediation in developing countries and plays a leading role in technology engineering radiation effect.

De novo transcriptome sequencing and analysis of male and female swimming crab (Portunus trituberculatus) reproductive systems during mating embrace (stage II).

BACKGROUND: The swimming crab Portunus trituberculatus is one of the most commonly farmed crustaceans in China. As one of the most widely known and high-value edible crabs, it crab supports large crab fishery and aquaculture in China. Only large and sexually mature crabs can provide the greatest economic benefits, suggesting the considerable effect of reproductive system development on fishery. Studies are rarely conducted on the molecular regulatory mechanism underlying the development of the reproductive system during the mating embrace stage in this species. In this study, we used high-throughput sequencing to sequence all transcriptomes of the P. trituberculatus reproductive system. RESULTS: Transcriptome sequencing of the reproductive system produced 81,688,878 raw reads (38,801,152 and 42,887,726 reads from female and male crabs, respectively). Low-quality (quality <20) reads were trimmed and removed, leaving only high-quality reads (37,020,664 and 41,021,030 from female and male crabs, respectively). A total of 126,188 (female) and 164,616 (male) transcripts were then generated by de novo transcriptome assembly using Trinity. Functional annotation of the obtained unigenes revealed that a large number of key genes and some important pathways may participate in cell proliferation and signal transduction. On the basis of our transcriptome analyses and as confirmed by quantitative real-time PCR, a number of genes potentially involved in the regulation of gonadal development and reproduction of P. trituberculatus were identified: ADRA1B, BAP1, ARL3, and TRPA1. CONCLUSION: This study is the first to report on the whole reproductive system transcriptome information in stage II of P. trituberculatus gonadal development and provides rich resources for further studies to elucidate the molecular basis of the development of reproductive systems and reproduction in crabs. The current study can be used to further investigate functional genomics in this species.

Bacterial communities in co-cultured fish intestines and rice field soil irrigated with aquaculture wastewater.

In some regions, integrated rice-fish farms have been developed to balance the needs of aquaculture wastewater discharge and rice field irrigation. In this type of aqua-agriculture system, soil is irrigated with aquaculture wastewater, and intestinal bacteria in cultured fish species likely impact soil bacteria through irrigation. However, little is known about the relationship between soil bacteria and intestinal bacteria in some carp species commonly co-cultured in some Asian regions. Therefore, we co-cultured five carp species in aquaculture ponds and used the aquaculture wastewater to irrigate rice fields for over 5 years, and then compared carp intestinal bacterial communities with rice field soil bacterial communities. The results from analysis of similarity and SourceTracker analysis showed that a low similarity (R = 0.7908, P = 0.001) and contribution (an average of 9.9% of bacterial genera) of intestinal bacteria to soil bacterial communities although 77.5% of soil bacterial genera were shared by intestinal bacteria. Our results also indicated that intestinal bacteria in the numerically dominant fish species in the co-culture system do not necessarily impact soil bacteria more significantly than those of less abundant carp species, and that intestinal bacterial communities in one single fish species may impact certain soil bacterial phyla more significantly than others. Our results provide a better understanding of the impact of aquaculture wastewater on rice fields and will be helpful for the development of this type of aqua-agriculture system.

Monitoring, modeling and projection of harmful algal blooms in China.

This review assesses monitoring, modeling and projection of harmful algal blooms (HABs) in China, with a focus on near-term warning and long-term trend projection. We reviewed phytoplankton monitoring as early warning, remote sensing for offshore environments, building models to describe ecological processes, modeling and forecasting near-term or seasonal HAB events, and projection of long-term HAB trend in China. Over the past 40 years, great progresses were made in traditional observation capability of HABs, and some reliable remote sensing algorithms were developed for HABs in optically complex coastal waters in Chinese seas. Numerical models have been applied to simulating real-world algal bloom events successfully; and these models, to some degree, are capable of predicting the time and place of HAB occurrence. In terms of long-term trend, HABs appeared to have shown diversified forms, being more miniaturized and more harmful. The development of an integrated monitoring and early-warning system of algal blooms and HABs should be a necessary first step to provide an effective management tool for mitigating damages associated with the occurrence of HABs in China.

A high abundance of Firmicutes in the intestine of chinese mitten crabs (Eriocheir sinensis) cultured in an alkaline region.

The Chinese mitten crab (Eriocheir sinensis) is a popular aquaculture product in East Asia, especially in China. In the last decade, rice-crab co-culture has rapidly expanded in China. Under this model, crabs are raised in rice fields instead of in traditional aquaculture ponds. In this study, we cultured two varieties of Chinese mitten crabs (Changjiang and Liaohe) in an alkaline region in northwest China and used Illumina MiSeq sequencing to compare the intestinal bacterial alpha diversity and community structure between traditional and co-culture aquaculture models, between two crab varieties, and between female and male crabs. Significant variations in intestinal bacterial communities were found between crab varieties and between female and male crabs but not between aquaculture models. These results show that rice-crab co-culture operations did not obviously impact the crab intestinal bacterial community compared with traditional pond aquaculture. Firmicutes was the most abundant bacterial phylum in the crab intestines (78%, relative abundance). Three dominant operational taxonomic units (OTUs) represented 73.2% of Firmicutes sequences and 56.8% of all sequences. A dominant OTU assigned as Firmicutes that was negatively correlated with crab body length, width, and weight was found in the source water for the experimental area. The results of this study suggest that the aquaculture of Chinese mitten crabs in alkaline regions requires more study to improve cultivation techniques.

Hydrogen gas alleviates acute alcohol-induced liver injury by inhibiting JNK activation.

Binge alcohol drinking is fast becoming a global health concern, with the liver among the first organ involved and the one afflicted with the greatest degree of injury. Oxidative stress, alterations in hepatic metabolism, immunity and inflammation have all been reported to contribute to the development of alcoholic liver disease (ALD). Hydrogen gas (H2) serves a key role in the modulation of hepatic redox, immune and inflammatory homeostasis. However, the effects of treatment using intraperitoneal injection of H2 on ALD remain unexplored. Therefore, the aim of the present study was to investigate the effects and underlying mechanism of intraperitoneal injection of H2 on acute alcohol-induced liver injury in a mouse model. H2 was administered by daily intraperitoneal injections (1.0 ml/100 g) for 4 days. On day 4, the mice received H2 after fasting for 5.5 h. After 30 min, the mice were administered with 33% (v/v) ethanol at a cumulative dose of 4.5 g/kg body weight by four equally divided gavages at 20-min intervals. Blood and liver tissues were collected at 16 h after the first ethanol gavage. Subsequently, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride and total cholesterol (TC) levels were analyzed using an Automatic Clinical Analyzer. Hepatic JNK activity and GAPDH levels were examined by western blotting. It was observed that acute ethanol gavage induced liver injury, as indicated by significantly increased serum ALT and AST levels, which were effectively decreased by H2 at 16 h after the first ethanol gavage. In addition, H2 treatment reduced serum TC levels in the Alcohol+H2 group when compared with those in Alcohol group. Mechanistically, H2 attenuated hepatic JNK phosphorylation induced by acute ethanol gavage. Therefore, the results of the present study demonstrated that treatment with exogenous H2 by intraperitoneal injection may alleviate acute alcohol-induced liver injury by inhibiting hepatic JNK activation, which may represent a novel therapeutic strategy for ALD.

Reproductive performance and semen characteristics of pond-reared and wild-caught large-sized male broodstock of the Chinese mitten crab Eriocheir sinensis.

There has been a recent emphasis on production of large-sized Eriocheir sinensis broodstock. In China, aquaculturists generally prefer wild-caught (WC) crabs from the Yangtze River as broodstock because offspring performance is superior to that of pond-reared (PR) broodstock. Currently, however, there is a ban on fishing in the Yangtze River, and effects on E. sinensis breeding have not been ascertained. There was comparison in the present study of reproductive performance and semen characteristics of male broodstock of PR and WC groups. After copulation, sperm quantity in the vas deferens of crabs in specimens of both groups was large, although there was a consistent decrease in vaso-somatic index. Although sperm density of PR crabs was less, that of WC specimens remained relatively constant. Specimens of neither group, however, had changes in the hepatopancreas index or condition factor, and sperm survival was close to 100%. Although the acrosome reaction was detected in response to cold-temperature induction, there were differences in extent of reaction to cold temperatures. Importantly, in as many as 98% of sperm from female spermathecae, the reaction was completed, which was considerably greater than 15% for sperm of males post-mating. It is concluded there was no difference between PR and WC crabs with respect to reproductive performance or semen characteristics, and, notably, sperm from PR crabs were of sufficient quality for use in E. sinensis aquaculture enterprises. Accordingly, it is predicted the Yangtze River fishing ban would only have a limited effect on supply of male E. sinensis broodstock.

Alkaline phosphatase activity during a phosphate replete dinoflagellate bloom caused by Prorocentrum obtusidens.

Prorocentrum obtusidens Schiller (formerly P. donghaiense Lu), a harmful algal species common in the East China Sea (ECS), often thrives with the depletion of phosphate. Three cruises in the spring of 2013 sampled an entire P. obtusidens bloom process to investigate the dynamics of alkaline phosphatase activity (APA) and phosphorus (P) status of the bloom species using both bulk and cell-specific assays. Unlike previous studies, the bloom of P. obtusidens occurred in a phosphate replete environment. Very high APA, with an average of 76.62 ± 90.24 nmol L-1 h-1, was observed during the early-bloom phase, a value comparable to that in low phosphate environments. The alkaline phosphatase (AP) hydrolytic kinetics also suggested a more efficient AP system with a lower half-saturation constant (Ks), but higher maximum potential hydrolytic velocity (Vmax) in this period. The APA decreased significantly with an average of 24.98 ± 30.98 nmol L-1 h-1 when the bloom reached its peak. The lack of a correlation between dissolved inorganic phosphate (DIP) or dissolved organic phosphate (DOP) concentration and APA suggested that the APA was regulated by the internal P growth demand, rather than the external P availability during the phosphate replete P. obtusidens bloom. These findings facilitate an understanding of the P. obtusidens acclimation strategy with respect to P variations in terms of AP expression during blooms in the ECS.

Down-regulation of telomerase activity and activation of caspase-3 are responsible for Tanshinone I-induced apoptosis in monocyte leukemia cells in vitro.

Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR-enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells.

Si-Wu-Tang Alleviates Nonalcoholic Fatty Liver Disease via Blocking TLR4-JNK and Caspase-8-GSDMD Signaling Pathways.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) has high global prevalence; however, the treatments of NAFLD are limited due to lack of approved drugs. METHODS: Mice were randomly assigned into three groups: Control group, NAFLD group, NAFLD plus Si-Wu-Tang group. A NAFLD mice model was established by feeding with a methionine- and choline-deficient (MCD) diet for four weeks. Si-Wu-Tang was given orally by gastric gavage at the beginning of 3rd week, and it lasted for two weeks. The treatment effects of Si-Wu-Tang were confirmed by examining the change of body weight, serum alanine aminotransferase (ALT) and aspartate transaminase (AST) levels, Oil Red O staining, and hematoxylin and eosin (H&E) staining of the liver samples and accompanied by steatosis grade scores. The expression and activation of the possible signaling proteins involved in the pathogenesis of NAFLD were determined by western blotting. RESULTS: Mice fed with four weeks of MCD diet displayed elevated serum levels of ALT and AST, while there was decreased body weight. The hepatic Oil Red O staining and H&E staining showed severe liver steatosis with high steatosis grade scores. All these can be improved by treating with Si-Wu-Tang for two weeks. Mechanistically, the increased hepatic TLR4 expression and its downstream JNK phosphorylation induced by MCD diet were suppressed by Si-Wu-Tang. Moreover, the upregulations of Caspase-8, gasdermin D (GSDMD), and cleaved-GSDMD in liver mediated by MCD diet were all inhibited by Si-Wu-Tang. CONCLUSIONS: Treatment with Si-Wu-Tang improves MCD diet-induced NAFLD in part via blocking TLR4-JNK and Caspase-8-GSDMD signaling pathways, suggesting that Si-Wu-Tang has potential for clinical application in treating NAFLD.

Therapeutic effect of traditional Chinese medicine on coagulation disorder and accompanying intractable jaundice in hepatitis B virus-related liver cirrhosis patients.

AIM: To observe the therapeutic effects of new traditional Chinese medicine (TCM) therapy on coagulation disorder and accompanying intractable jaundice in HBV-related liver cirrhosis patients. METHODS: Using stratified random sampling according to fibrinogen (Fib) levels, 145 liver cirrhosis patients due to hepatitis B complicated by coagulation disorder were treated. Of them, 70 in research group were treated with TCM by "nourishing yin, cooling blood and invigorating blood circulation" and Western medicine, 75 in control group were treated with conventional Western medicine. The indexes of liver function, coagulation function and bleeding events were observed and compared. RESULTS: The prothrombin time (PT) was shorter and the fibrinogen (Fib) level was higher in the research group than in the control group (Fib = 1.6-2.0 g/L, 1.1-1.5 g/L, and < or = 1.0 g/L). The total bilirubin (TBIL) level was significantly lower in the research group than in the control group, except for the subgroup of FIB < or = 1.0 g/L. CONCLUSION: TCM therapy can improve coagulation fuction and decrease TBIL.

Traditional Chinese medicine syndromes of chronic hepatitis B with precore mutant.

AIM: This study aims at exploring the distribution of TCM syndromes in CHB patients with HBV pre-core mutation (1896) and the relationship between pre-core mutation and T lymphocytes subgroup, through which to provide objective data on clinical syndrome differentiation of TCM, and further to suggest the therapeutic principle and guide clinical treatment. METHODS: One hundred and forty CHB patients were evenly divided into two study groups, HBV pre-core mutant group and HBV pre-core wild-type group. Besides, 30 healthy blood donors were selected as a healthy control group. HBV-labeled compound, T lymphocytes subgroup, and HBV-DNA pre-core mutant were tested in the study groups. T lymphocytes subgroup were also tested in the control group. All the patients were both diagnosed by syndrome differentiation of TCM and western medicine. RESULTS: The most common syndrome in mutant group was damp-heat combined with blood stasis, and the most common syndrome in the wild-type group was damp-heat stasis in the middle-jiao. There were more cases of medium and severe hepatitis in mutant group than that in wild-type group. The content of CD4+ lymphocytes and CD4+/CD8+ ratio were decreased gradually (healthy control group>wild-type group>mutant group). In the wild-type group, severe and medium CHB patients had considerably lower level of them than mild CHB patients. However, in the mutant group, the opposite result appeared. Meanwhile, the content of HBV-DNA in mutant group was higher than that in wild-type group. CONCLUSION: Damp, heat, toxin and blood stasis were the basic pathogens of CHB, whether pre-core mutant or not. CHB with precore mutant may lead to more severe hepatitis. The decreased content of CD4+ lymphocytes and ratio of CD4+/CD8+ may be taken as one of the indices in confirming the deficiency syndrome of CHB patients with pre-core mutation.

Chinese medicine syndrome distribution of chronic hepatitis B virus carriers in immunotolerant phase.

OBJECTIVE: To explore Chinese medicine (CM) syndrome distribution of chronic hepatitis B virus (HBV) carriers in immunotolerant phase (ITP). METHODS: One hundred and eighty-five chronic HBV carriers in ITP, seen in the Third Affiliated Hospital of Sun Yat-sen University from May 2009 to December 2010, were admitted in an observational study under the guidance of CM. Patients' CM symptoms and signs, demographics, liver biochemistries, and qualitative HBV DNA were recorded in the questionnaires. CM syndromes were then differentiated to 15 detailed types and analyzed by generalization. Lastly, the location, pathogenic factors and nature of the disease were also assessed. RESULTS: When CM syndrome patterns were differentiated to 15 types, there were 27 (15%) no syndrome cases, 94 (50%) single syndrome cases and 64 (35%) compound syndromes cases. The main detailed syndromes included Liver (Gan)-qi depression (LQD), Kidney (Shen)-qi deficiency (KQD), Spleen (Pi)-qi deficiency (SQD) and Kidney-yang deficiency (KYAD). After CM syndromes generalized to five types, their frequency was Spleen-Kidney deficiency (SKD)>LQD>inner dampness-heat retention (IDHR)>Liver-Kidney deficiency (LKD)>blood stasis blocking collateral (BSBC). SKD and LQD occupied 64%. The disease location included Liver, Gallbladder (Dan), Spleen, Stomach (Wei) and Kidney. The pathogenic factors were mainly qi stagnation, qi deficiency, yang deficiency, concurrently dampness-heat and blood stasis. The deficiency syndrome was more than excess syndrome in its nature. CONCLUSIONS: Most of chronic HBV carriers in ITP have their CM syndrome, and the most common types are SKAD, LQD. This study suggests that the natural history may be improved through breaking the state of immune tolerance or shorten the time of ITP by strengthening Spleen-Kidney and reliving Liver qi.

The correlation between Prorocentrum donghaiense blooms and the Taiwan warm current in the East China Sea - evidence for the "Pelagic Seed Bank" hypothesis.

During the last two decades, large-scale high biomass algal blooms of the dinoflagellate Prorocentrum donghaiense Lu have occurred frequently in the East China Sea (ECS). The role of increasing nutrient concentrations in driving those blooms is well-established, but the source population that initiates them is poorly understood. We hypothesized that the front of Taiwan Warm Current (TWC) may serve as a 'seed bank' that initiates P. donghaiense blooms in the ECS, as the physiochemical conditions in the TWC are suitable for the growth of P. donghaiense. In order to test this hypothesis, two surveys at different spatio-temporal scales were conducted in 2010 and 2011. We found a strong correlation in space and time between the abundance of P. donghaiense and the TWC. The spatial extent of the P. donghaiense bloom coincided with the TWC front in both 2010 and 2011. During the early development of the blooms, P. donghaiense concentration was highest at the TWC front, and then the bloom mass shifted inshore over the course of our 2011 survey. The TWC also moved inshore, albeit after the appearance of P. donghaiense. Overall, these results support our hypothesis that P. donghaiense blooms develop from the population at the TWC front in the ECS, suggesting the role of the ocean current front as a seed bank to dinoflagellate blooms.

[Relationship between traditional Chinese medicine syndrome types and serum HBV DNA level in chronic HBV carriers positive for HBeAg].

OBJECTIVE: To explore the relationship between the syndrome types in traditional Chinese medicine (TCM) and serum HBV DNA load in chronic HBV carriers positive for HBeAg. METHODS: According to the TCM syndrome types, 185 chronic HBV carriers with HBeAg positivity were classified into single syndrome group (liver Qi depression, kidney Qi deficiency, spleen Qi deficiency, and kidney Yang deficiency), compound syndrome group, and unidentifiable syndrome group; based on the nature of the condition in TCM terms, the patients were classified into excess syndrome group, deficiency syndrome group and comorbidity syndrome group. The serum HBV DNA levels in these cases were analyzed in relation to the TCM syndrome types and disease nature. RESULTS: HBV DNA levels showed no significant difference among the patients with single syndrome, compound syndromes and unidentifiable syndrome (F=0.910, P=0.404), nor among the patients with the 5 different single TCM syndromes (χ²=4.672, P=0.323) or those with different disease nature (F=0.631, P=0.596). CONCLUSION: Serum HBV DNA level can not be considered as the evidence for syndrome differentiation in chronic HBV carriers with positive HBeAg.