Ma Xing Shi Gan Decoction Protects against PM2.5-Induced Lung Injury through Suppression of Epithelial-to-Mesenchymal Transition (EMT) and Epithelial Barrier Disruption.

This research was designed to explore the effect of Ma Xing Shi Gan decoction (MXD) in alleviating particulate matter less than 2.5 µm in diameter (PM2.5) induced lung injury from the perspective of epithelial barrier protection and inhibition of epithelial-to-mesenchymal transition (EMT). Rats were exposed to PM2.5 to establish a lung injury model in vivo, and a PM2.5-stimulated primary cultured type II alveolar epithelial cell model was introduced in vitro. Our results indicated that MXD alleviated the weight loss and pathologic changes and improved the epithelial barrier dysfunction. MXD also significantly inhibited the TGF-ß/Smad3 pathway, increased the level of ZO-1 and claudin-5, and reversed the EMT process. Notably, the protection of MXD was abolished by TGF-ß in vitro. Our results indicated that MXD has a protection against PM2.5-induced lung injury. The proposed mechanism is reversing PM2.5-induced EMT through inhibiting TGF-ß/Smad3 pathway and then upregulating the expression of tight-junction proteins.

Surveillance-based evidence: elimination of schistosomiasis as a public health problem in the Peoples' Republic of China.

BACKGROUND: A steady progress on schistosomiasis control in the Peoples' Republic of China (P.R. China) was achieved and broadened into the twelve-year medium and long term national plan (MLNP) which marled the implementation of an integrated control strategy across all endemic areas in P.R. China in 2004. To understand the endemic trends of schistosomiasis to assess the effectiveness of an integrated strategy, we conducted an analysis of schistosomiasis surveillance data spanned from 2005 to 2015. METHODS: The schistosomiasis sentinel surveillance data from sentinel sites were collected and analyzed from 2005 to 2015. In these sentinel sites, residents aged 6 years or above were screened annually by indirect hemagglutination assay (IHA), while only antibody positives were followed by stool examination either Kato-katz method (KK) and/or hatching technique (HT). Domestic animals raised in sentinel sites were examined by HT for confirming the infection of schistosomes. Snail investigation was conducted each year through systematic sampling method combined with environmental sampling method. The snails collected from field were tested by microscopic dissection method. The infection rates of schistosomes in residents, domestic animals and snails, as well as the indicators reflecting the snails' distribution were calculated and analyzed. ANOVA analysis was used to examine the changes of the number of eggs per gram feces in population and Chi-square test was used to examine any change in proportions among groups. RESULTS: A total of 148 902 residents from sentinel sites attended this study and 631 676 blood samples were examined by IHA test during the 11 covered years. The annual average antibody positive rates presented a significant decrease trends, from 17.48% (95% CI: 17.20-17.75%) in 2005 to 5.93% (95% CI: 5.71-6.15%) (χ2 = 8890.47, P < 0.001) in 2015. During 2005-2015, the average infection rate of schistosomes in residents declined from 2.07% (95% CI: 1.96-2.17%) to 0.13% (95% CI: 0.09-0.16%), accompanied by significant decrease of infection intensity in population. In 2015, the stool positives were only found in farmers, fishermen and boatmen with infection rate of 0.16% (95% CI: 0.11-0.20%), 0.17% (95% CI: 0-0.50%) respectively. The infection rate of schistosomes in domestic animals dropped from 9.42% (538/5711, 95% CI: 8.66-10.18%) to 0.08% (2/2360, 95% CI: 0-0.20%) from 2005 to 2015. Infections were found in eight species of domestic animals at the beginning of surveillance while only two cattle were infected in 2015. Totally 98 ha of new snail habitats were found, while 94.90% (93/98) distributed in lake and marshland regions. The percentage of frames with snails decreased from 16.96% (56 884/33 5391, 95% CI: 16.83-17.09%) in 2005 to 4.28% (18 121/423 755, 95% CI: 4.22-4.34%) in 2014, with a slightly increase in 2015. Meanwhile, the infection rate of schistosomes in snails was decreased from 0.26% (663/256 531, 95% CI: 0.24-0.28%) to zero during 2005-2015. CONCLUSIONS: The infection rate of schistosomes declined significantly, providing evidence that the goal of the MLNP was achieved. Elimination of schistosomiasis as a public health problem defined as WHO was also reached in P.R. China nationwide. Surveillance-response system should be improved and strengthened to realize the final goal of schistosomiasis elimination.

Targeting of TLE3 by miR-3677 in human breast cancer promotes cell proliferation, migration and invasion.

Numerous studies have indicated an important function of microRNAs (miRs) in breast cancer (BC) progression, oncogenesis and metastasis. However, the function of miR-3677, which has been revealed to be upregulated in BC [The Cancer Genome Atlas (TCGA) data], has not been investigated to date. In the present study, miR-3677 was revealed to be upregulated in BC as determined using TCGA. miR-3677 was significantly upregulated in BC tissues and cell lines compared with those noted in adjacent non-cancerous tissues and primary normal breast cells (P<0.05). The overexpression of miR-3677 promoted the cell proliferation, migration and invasion of BC cells. Using bioinformatics algorithms and luciferase assays, a novel target gene for miR-3677, namely transducin-like enhancer of Split3 (TLE3), was identified. Silencing of TLE3 in miR-3677-transfected BC cells suppressed their proliferation and migration. An inverse correlation was observed between miR-3677 and TLE3 expression levels in human BC tissues. In conclusion, the present study demonstrated that miR-3677 promoted BC cell proliferation, migration and invasion by inhibiting TLE3 expression, which provided a novel mechanism and a promising therapeutic target for patients with BC.

Ma Xing Shi Gan Decoction Attenuates PM2.5 Induced Lung Injury via Inhibiting HMGB1/TLR4/NFκB Signal Pathway in Rat.

Ma Xing Shi Gan Decoction (MXD), a classical traditional Chinese medicine prescription, is widely used for the treatment of upper respiratory tract infection. However, the effect of MXD against particulate matters with diameter of less than 2.5 µm (PM2.5) induced lung injury remains to be elucidated. In this study, rats were stimulated with PM2.5 to induce lung injury. MXD was given orally once daily for five days. Lung tissues were harvested to assess pathological changes and edema. Myeloperoxidase (MPO) activity and malonaldehyde (MDA) content in lung were determined to evaluate the degree of injury. To assess the barrier disruption, the bronchoalveolar lavage fluid (BALF) was collected to determine the total protein content and count the number of neutrophils and macrophages. For evaluating the activation of macrophage in lung tissue, CD68 was detected using immunohistochemistry (IHC). The levels of inflammatory factors including tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1ß), and interleukin-6 (IL-6) in BALF and serum were measured. In vitro, a PM2.5-activated RAW 264.7 macrophages inflammatory model was introduced. To evaluate the protective effect of MXD-medicated serum, the cell viability and the release of inflammatory factors were measured. The effects of MXD on the High mobility group box-1/Toll-like receptor 4/Nuclear factor-kappa B (HMGB1/TLR4/NFκB) pathway in lung tissue and RAW 264.7 cells were assessed by Western blot. For further confirming the protective effect of MXD was mediated by inhibiting the HMGB1/TLR4/NFκB pathway, RAW 264.7 cells were incubated with MXD-medicated serum alone or MXD-medicated serum plus recombinant HMGB1 (rHMGB1). MXD significantly ameliorated the lung injury in rats, as evidenced by decreases in the pathological score, lung edema, MPO activity, MDA content, CD68 positive macrophages number, disruption of alveolar capillary barrier and the levels of inflammatory factors. In vitro, MXD-medicated serum increased cell viability and inhibited the release of inflammatory cytokines. Furthermore, MXD treatment was found to inhibit HMGB1/TLR4/NFκB signal pathway both in vivo and in vitro. Moreover, the protection of MXD could be reversed by rHMGB1 in RAW 264.7. Taken together, these results suggest MXD protects rats from PM2.5 induced acute lung injury, possibly through the modulation of HMGB1/TLR4/NFκB pathway and inflammatory responses.

Efficient Secretion of the ß-Galactosidase Bgal1-3 via both Tat-Dependent and Tat-Independent Pathways in Bacillus subtilis.

In this study, the twin-arginine (Tat) signal peptide PhoD was used to direct the secretion of the ß-galactosidase Bgal1-3 into the growth medium of an engineered strain of Bacillus subtilis 168. After 24 h of cultivation, the extracellular activity reached 1.15 U/mL, representing 78% of the total activity. Bgal1-3 was exported via both Tat-dependent and Tat-independent pathways. To improve the secretion amounts, two more copies of the target gene were inserted into the designated loci on the chromosome, further improving the extracellular enzymatic activity to 2.15 U/mL. The engineered strain with three copies of bgal1-3 was genetically stable after 150 generations. To the best of our knowledge, this is the first report on the functional secretion of a heterologous protein via both Tat-dependent and Tat-independent pathways mediated by a Tat signal peptide in B. subtilis. Furthermore, this study provides us with a markerless engineered strain for the production of ß-galactosidase.

Engineering a novel glucose-tolerant ß-glucosidase as supplementation to enhance the hydrolysis of sugarcane bagasse at high glucose concentration.

BACKGROUND: Most ß-glucosidases reported are sensitive to the end product (glucose), making it the rate limiting component of cellulase for efficient degradation of cellulose through enzymatic route. Thus, there are ongoing interests in searching for glucose-tolerant ß-glucosidases, which are still active at high glucose concentration. Although many ß-glucosidases with different glucose-tolerance levels have been isolated and characterized in the past decades, the effects of glucose-tolerance on the hydrolysis of cellulose are not thoroughly studied. RESULTS: In the present study, a novel ß-glucosidase (Bgl6) with the half maximal inhibitory concentration (IC 50) of 3.5 M glucose was isolated from a metagenomic library and characterized. However, its poor thermostability at 50 °C hindered the employment in cellulose hydrolysis. To improve its thermostability, random mutagenesis was performed. A thermostable mutant, M3, with three amino acid substitutions was obtained. The half-life of M3 at 50 °C is 48 h, while that of Bgl6 is 1 h. The K cat/K m value of M3 is 3-fold higher than that of Bgl6. The mutations maintained its high glucose-tolerance with IC 50 of 3.0 M for M3. In a 10-h hydrolysis of cellobiose, M3 completely converted cellobiose to glucose, while Bgl6 reached a conversion of 80 %. Then their synergistic effects with the commercial cellulase (Celluclast 1.5 L) on hydrolyzing pretreated sugarcane bagasse (SCB) were investigated. The supplementation of Bgl6 or mutant M3 to Celluclast 1.5 L significantly improved the SCB conversion from 64 % (Celluclast 1.5 L alone) to 79 % (Bgl6) and 94 % (M3), respectively. To further evaluate the application potential of M3 in high-solids cellulose hydrolysis, such reactions were performed at initial glucose concentration of 20-500 mM. Results showed that the supplementation of mutant M3 enhanced the glucose production from SCB under all the conditions tested, improving the SCB conversion by 14-35 %. CONCLUSIONS: These results not only clearly revealed the significant role of glucose-tolerance in cellulose hydrolysis, but also showed that mutant M3 may be a potent candidate for high-solids cellulose refining.

[Effect of ditching for drain on control of Oncomelania hupensis snail in beaches of Dongting Lake].

OBJECTIVE: To evaluate the effect of ditching for drain on the control of the breed of Oncomelania hupensis snails in beaches of Dongting Lake. METHODS: From November, 2009 to November, 2012, an 0. hupensis snail infested beach of the Yueyang jail and an O. hupensis snail infested beach of Junshan District were selected as research fields in the eastern Dongting Lake area, and the former, as the intervention field, was performed with the ditching for drain by excavators and the latter, as the control field, was not. RESULTS: Before the project implemented, the average soil moisture contents on the beaches in dry seasons of the two fields were both about 35.56%. After the project implemented, in the intervention field, the average soil moisture content was 26.53% which was significantly lower than that (35.56%) in the control field (F = 6.53, P < 0.05). The underground water levels in different heights in the intervention field were lower than those in the control field (χ2 = 33.33, P < 0.05). Before the project implemented, the natural death rates of the snails were 0.98% and 0.89% in the two research fields respectively (P > 0.05), and after the project implemented (in 2012), no adult and young snails were found in the interventional field, but in the control field, the average densities of living snails and young snails were 29.37 snails/0.1 m2 and 213 ± 108.45 snails/0.1 m2 respectively. CONCLUSION: The intervention of ditching for drain can decrease the soil moisture contents quickly and change the ecological condition, therefore, can control the breed of O. hupensis snails in the beaches of Dongting Lake.

Immunohistochemical analysis of phosphorylated mammalian target of rapamycin and its downstream signaling components in invasive breast cancer.

The present study aimed to investigate whether the mammalian target of rapamycin (mTOR) signaling pathway is activated in invasive breast cancer. The expression levels of phosphorylated (p)­mTOR at ser2448 were detected, as well as the expression levels of its downstream signaling molecules: Eukaryotic translation initiation factor 4E­binding protein 1 (4E­BP1), and p70 ribosomal protein S6 kinase 1 (S6K1). The correlation between p­mTOR, p­4E­BP1, p­S6K1, and the clinicopathological parameters of breast cancer were also determined. p­mTOR, p­4E­BP1 and p­S6K1 expression was detected in 285 breast cancer tumor samples and adjacent normal tissue samples using immunohistochemistry. The expression levels and the location of the proteins were analyzed and compared in the various tissue samples. Multivariate Cox regression was used to analyze the clinicopathological factors and prognosis associated with the tissue samples. The disease­free survival rate was examined using survival analyses and Log­rank tests. The results of the present study indicated that the expression levels of p­mTOR, p­4E­BP1, and p­S6K1 were significantly higher in breast cancer tissue, as compared with normal tissue (P<0.01). p­mTOR was predominantly expressed in the cytoplasm, whereas p­4E­BP1 and p­S6K1 were predominantly co­expressed in the cytoplasm and the nucleus. In addition, p­4E­BP1 and p­S6K1 were more likely to be expressed in the cytoplasm in breast cancer tissue samples, as compared with normal tissue samples (P<0.001). Positive p­mTOR was not significantly correlated with positive p­4E­BP1 and p­S6K1 expression. The survival analyses of the patients with positive p­mTOR, p­4E­BP1, and p­S6K1 tissue samples were not significantly different from those of the patients with negative tissue samples (P>0.05). Thus suggesting that these markers are not adequate risk factors for disease free survival (P>0.05). In conclusion, the results of the present study suggested that p­mTOR, p­4E­BP1, and p­S6K1 are activated in invasive breast cancer. In addition, the exclusive expression of p­4E­BP1 and p­S6K1 in the cytoplasm may be characteristic of progressive breast cancer. However, p­mTOR, p­4E­BP1, and p­S6K1 are not prognostic factors for breast cancer.

Three Gorges Dam: Impact of Water Level Changes on the Density of Schistosome-Transmitting Snail Oncomelania hupensis in Dongting Lake Area, China.

BACKGROUND: Schistosomiasis remains an important public health issue in China and worldwide. Oncomelania hupensis is the unique intermediate host of schistosoma japonicum, and its change influences the distribution of S. japonica. The Three Gorges Dam (TGD) has substantially changed the ecology and environment in the Dongting Lake region. This study investigated the impact of water level and elevation on the survival and habitat of the snails. METHODS: Data were collected for 16 bottomlands around 4 hydrological stations, which included water, density of living snails (form the Anxiang Station for Schistosomiasis Control) and elevation (from Google Earth). Based on the elevation, sixteen bottomlands were divided into 3 groups. ARIMA models were built to predict the density of living snails in different elevation areas. RESULTS: Before closure of TGD, 7 out of 9 years had a water level beyond the warning level at least once at Anxiang hydrological station, compared with only 3 out of 10 years after closure of TGD. There were two severe droughts that happened in 2006 and 2011, with much fewer number of flooding per year compared with other study years. Overall, there was a correlation between water level changing and density of living snails variation in all the elevations areas. The density of living snails in all elevations areas was decreasing after the TGD was built. The relationship between number of flooding per year and the density of living snails was more pronounced in the medium and high elevation areas; the density of living snails kept decreasing from 2003 to 2014. In low elevation area however, the density of living snails decreased after 2003 first and turned to increase after 2011. Our ARIMA prediction models indicated that the snails would not disappear in the Dongting Lake region in the next 7 years. In the low elevation area, the density of living snails would increase slightly, and then stabilize after the year 2017. In the medium elevation region, the change of the density of living snails would be more obvious and would increase till the year 2020. In the high elevation area, the density of living snails would remain stable after the year 2015. CONCLUSION: The TGD influenced water levels and reduced the risk of flooding and the density of living snails in the study region. Based on our prediction models, the density of living snails in all elevations tends to be stabilized. Control of S. japonica would continue to be an important task in the study area in the coming decade.

Characterization of the cross-linked enzyme aggregates of a novel ß-galactosidase, a potential catalyst for the synthesis of galacto-oligosaccharides.

A novel ß-galactosidase (Bgal1-3) was isolated from a marine metagenomic library and then its cross-linked enzyme aggregates (CLEAs) were prepared. The enzymatic properties of Bgal1-3-CLEAs were studied and compared with that of the free enzyme. The thermostability and storage stability of Bgal1-3 were significantly improved after it was immobilized as CLEAs. The galactose-tolerance of the enzyme was also enhanced after the immobilization, which could relieve the inhibitory effect and then tends to be beneficial for the galacto-oligosaccharides (GOS) synthesis. Moreover, higher GOS yield was achieved (59.4 ± 1.5%) by Bgal1-3-CLEAs compared to the free counterpart (57.1 ± 1.7%) in an organic-aqueous biphasic system. The GOS content and composition of the syrups synthesized by the free enzyme and Bgal1-3-CLEAs were similar and they both contained at least seven different oligosaccharides with the degree of polymerization (DP) ranging between 3 and 9. Furthermore, Bgal1-3-CLEAs maintained 82.1 ± 2.1% activity after ten cycles of reuse; the GOS yield of the tenth batch was 52.3 ± 0.3%, which was still higher than that of the most former reports. To the best of our knowledge, this is the first report on the GOS synthesis using CLEAs of ß-galactosidase in an organic-aqueous biphasic system. The study not only further expands the application scope of CLEA, but also provides a potential catalyst for the synthesis of GOS with low cost.

[Diagnosis and treatment of schistosomiasis japonica-concensuses among ex- perts in Hunan, Hubei and Jiangxi provinces].

In order to further standardize the diagnosis and treatment of schistosomiasis japonica in China, on the basis of evidence-based medicine, the experts on schistosomiasis control from Hunan, Hubei and Jiangxi provinces summarized their consensuses on the disease after the discussion on the current situation and progress of clinical diagnosis and treatment of schistosomiasis in China, with the reference to the Diagnostic Criteria for Schistosomiasis (WS261-2006), which aimed to establish the therapeutic standards or guideline of schistosomiasis in China.

[Appropriate technology: a driver to eliminate schistosomiasis in China. In- sights gained from the First Forum on Schistosomiasis Control in China].

This paper described the First Forum on Schistosomiasis Control in China. With "Appropriate technology is essential to schistosomiasis prevention and control" as the topic, the forum shared the main interventions and experiences for schistosomiasis control and elimination, exhibited the recent achievements gained from the development of appropriate technology for schistosomiasis control, showed the technical support of appropriate technology for schistosomiasis elimination, and proposed the priority for the future research and development of appropriate technology for schistosomiasis control and elimination in China. It is considered that the forum builds a bridge for communicating information between the administrative sectors and specialized prevention and control institutions, between research institutions and manufactures, and between experts/professors and grassroots professional staff, and provides a platform to communicate the national schistosomiasis control knowledge of China.

[Clinical manifestations and treatment of schistosomiasis hematobia].

Schistosomiasis hematobia is one major human schistosomiasis. The disease is endemic in Africa and Mediterra- nean region, and is the main cause of urogenital diseases. Although only Schistosoma japonicum is spreading across the Mainland China, now more schistosomiasis hematobia cases are reported among aid projects and migrant workers to Africa, with the economy development and the increasing degree of foreign exchanges. Meanwhile, the relevant clinical data of.schistosomiasis hematobia are rare in China. This article reviews the clinical manifestations and progress in diagnosis and treatment of the disease.

[Evaluation on effect of treatment and assistance to advanced schistosomiasis patients in Hunan Province from 2004 to 2013].

OBJECTIVE: To comprehensively evaluate the effect of the program of treatment and assistance to advanced schistosomiasis patients in Hunan Province from 2004 to 2013. METHODS: The fund investment of the program, the profits of hospitals and the improvement of the patients' health were investigated by data collection and questionnaire survey. The evaluation index system of treatment and assistance to advanced schistosomiasis in Hunan Province was constructed by the Delphi method and analytic hierarchy process, and the program was assessed comprehensively. RESULTS: The evaluation index system including 6 primary indices and 33 secondary indices was established. Among all the primary indices, the score of the treatment and assistance (22.25) was the highest, and that of the satisfaction assessment (8.15) was the lowest, and the score of the comprehensive assessment was 87.06. The average cure rate of the patients was 13.08% from 2004 to 2013. More than 60% of the patients' disease condition got better, and nearly 70% of the patients' psychological condition improved, and more than 70% of patients' self-help ability and social contact improved, as well as family happiness increased. In addition, the annual average cost for caretakers decreased by 2000 Yuan, and the profits of all the fixed-point hospitals for treatment and assistance increased. CONCLUSION: The effectiveness and efficiency of the treatment and assistance to advanced schistosomiasis patients in Hunan Province is obvious, and the government should continuously invest in the program.

[Curative effect of Ruangan pills in treatment of schistosomiasis liver fibrosis].

OBJECTIVE: To explore the efficacy, mechanism and safety of silibinin combined with Ruangan pills (a Chinese herbal preparation) in the treatment of schistosomiasis liver fibrosis. METHODS: A total of 200 patients with schistosomiasis liver fibrosis were randomly divided into a control group and a treatment group, and 100 patients in each group were respectively administered with oral silibinin alone and oral silibinin combined with Ruangan pills, respectively. The curative effects in the two groups were evaluated in 3 months, 6 months, 9 months and 12 months respectively. RESULTS: The common five clinical symptoms of schistosomiasis liver fibrosis patients significantly relieved in the treatment group 12 months after the therapy, and the total efficiency reached more than 75%, which were significantly higher than that in the control group. In the treatment group and the control group, there was no improvement in the liver B ultrasonic classification 3 months and 6 months after the therapy (P > 0.05); however, in 9 months and 12 months, the liver B ultrasonic classification in the treatment group was better than that in the control group (P < 0.05, P < 0.01, respectively). For the four serum indexes of liver fibrosis, there was no significant differences between the two groups in 3 months, however, in 6 months, 9 months, and 12 months, there was a significant improvement in the treatment group compared with the control group. There were no obviously adverse effects in two groups. CONCLUSION: Silibinin combined with Ruangan pills has a better curative effect in the treatment of schistosomiasis liver fibrosis.

Identification of optimum scopes of environmental factors for snails using spatial analysis techniques in Dongting Lake Region, China.

BACKGROUND: Owing to the harmfulness and seriousness of Schistosomiasis japonica in China, the control and prevention of S. japonica transmission are imperative. As the unique intermediate host of this disease, Oncomelania hupensis plays an important role in the transmission. It has been reported that the snail population in Qiangliang Lake district, Dongting Lake Region has been naturally declining and is slowly becoming extinct. Considering the changes of environmental factors that may cause this phenomenon, we try to explore the relationship between circumstance elements and snails, and then search for the possible optimum scopes of environmental factors for snails. METHODS: Moisture content of soil, pH, temperature of soil and elevation were collected by corresponding apparatus in the study sites. The LISA statistic and GWR model were used to analyze the association between factors and mean snail density, and the values in high-high clustered areas and low-low clustered areas were extracted to find out the possible optimum ranges of these elements for snails. RESULTS: A total of 8,589 snail specimens were collected from 397 sampling sites in the study field. Besides the mean snail density, three environmental factors including water content, pH and temperature had high spatial autocorrelation. The spatial clustering suggested that the possible optimum scopes of moisture content, pH, temperature of the soil and elevation were 58.70 to 68.93%, 6.80 to 7.80, 22.73 to 24.23°C and 23.50 to 25.97 m, respectively. Moreover, the GWR model showed that the possible optimum ranges of these four factors were 36.58 to 61.08%, 6.541 to 6.89, 24.30 to 25.70°C and 23.50 to 29.44 m, respectively. CONCLUSION: The results indicated the association between snails and environmental factors was not linear but U-shaped. Considering the results of two analysis methods, the possible optimum scopes of moisture content, pH, temperature of the soil and elevation were 58.70% to 68.93%, 6.6 to 7.0, 22.73°C to 24.23°C, and 23.5 m to 26.0 m, respectively. The findings in this research will help in making an effective strategy to control snails and provide a method to analyze other factors.

Identification and characterization of an unusual glycosyltransferase-like enzyme with ß-galactosidase activity from a soil metagenomic library.

Glycosyltransferases and glycoside hydrolases are two diversified groups of carbohydrate-active enzymes (CAZymes) in existence, they serve to build and break down the glycosidic bonds, respectively, and both categories have formed many sequence-based families. In this study, a novel gene (glyt110) conferring ß-galactosidase activity was obtained from a metagenomic library of Turpan Basin soil. Sequence analysis revealed that glyt110 encoded a protein of 369 amino acids that, rather than belonging to a family typically known for ß-galactosidase activity, belonged to glycosyltransferase family 4. Because of this unusual sequence information, the novel gene glyt110 was subsequently expressed in Escherichia coli BL21(DE3), and the recombinant enzyme (Glyt110) was purified and characterized. Biochemical characterization revealed that the ß-galactosidase activity of Glyt110 toward o-nitrophenyl-ß-D-galactopyranoside (ONPG) and lactose were identified to be 314±18.3 and 32±2.7 U/mg, correspondingly. In addition, Glyt110 can synthesize galacto-oligosaccharides (GOS) using lactose as substrate. A GOS yield of 47.2% (w/w) was achieved from 30% lactose solution at 50 °Ð¡, pH 8.0 after 10 h reaction. However, Glyt110 was unable to glycosylate either N-acetylated saccharides or lactose and galactose using UDP-gal as sugar donor, and its glycosyltransferase activity needs further investigation. These results indicated that Glyt110 is an unusual enzyme with ß-galactosidase activity but phylogenetically related to glycosyltransferase. Our findings may provide opportunities to improve the insight into the relationship between glycosyltransferases and glycoside hydrolases and the sequence-based classification.

[Schistosomiasis control effect of measures of replacing cattle with machine for cultivation and forbidding depasturage of livestock on marshlands in marshland and lake regions].

OBJECTIVE: To evaluate the schistosomiasis control effect of the measures of replacing cattle with machine for cultivation and forbidding depasturage of livestock on marshlands in marshland and lake regions. METHODS: The retrospective review and field survey were implemented in the Jiangling and Gongan counties of Hubei Province where were implemented with the measure of replacing cattle with machine for cultivation, and Yuanjiang and Huarong counties of Hunan Province where were implemented with the measure of forbidding the depasturage of livestock on marshlands. The schistosome infection status of human, cattle, and Oncomelania hupensis snails, and schistosome-infested field excreta were surveyed from 2007 to 2013. The effects of the interventions were compared before and after their implementations. RESULTS: The 6 villages of Hubei Province were implemented with the measure of replacing cattle with machine for cultivation, and 7 villages of Hunan Province were implemented with the measure of forbidding the depasturage of livestock on marshlands. From 2007 to 2013, the schistosome infection rates of residents declined from 3.95% to 0.70% (χ2 = 128.376, P < 0.05), with the descend range of 82.53%. The descend ranges of the measures of replacing cattle with machine for cultivation and forbidding the depasturage of livestock on marshlands were 83.081% and 81.62% respectively, and there was no significant difference between the two measures (χ2 = 0.132, P > 0.05). The infection rate of cattle decreased from 3.66% in 2007 to 0.65% in 2013, and the descend range was 82.24% (χ2 = 13.692, P < 0.05). The field excreta was surveyed in the snail breeding place in 2013. The investigated area was 157.435 hm2, and 625 samples of field feces of cattle were collected with the density of field excreta of 3.97/hm2, and the positive rate was 1.12% (7/625). The schistosome infection rate of snails and the density of infected snails decreased successively from 2007, and no infected snails were detected from 2011 to 2013. CONCLUSIONS: The schistosomiasis endemic situation are decreased significantly after the interventions of replacing cattle with machine for cultivation and forbidding the depasturage of livestock on marshlands. Therefore, schistosomiasis control effect of these measures is notable in the marshland and lake regions.

[Retrospective investigation of schistosomiasis endemic situation in Hunan Province].

OBJECTIVE: To understand the dynamic rules of schistosomiasis endemic situation before and after reaching the criteria of schistosomiasis transmission controlled or transmission interrupted, so as to provide the evidence for improving schistosomiasis control. METHODS: Wuling District, Xihu District and Linli County were selected and investigated retrospectively to collect the schistosomiasis epidemiological information 10 years before they reached the criteria of transmission controlled and the subsequent years until 2008. A database of retrospective investigation was established for analyzing the trends and rules of changes of the Oncomelania hupensis snail status and infection status of cattle and human. RESULTS: In Wuling District, the endemic situation was declining, and no schistosome infection persons, animals and snails were found after 1974. There was no rebound until 2008. In Xihu District, the endemic situation reached the criteria of transmission controlled in 1997, and the endemic situation was stable. The human infection rate was positively correlated with the area of infection snails (r = 0.584, P < 0.05). In Linli County, there were no snails, no infected persons and cattle twice, but 2 endemic rebounds, and there were positive correlations between the densities of living snails and the infection rates of human and animal during the endemic rebound period. CONCLUSION: The snail status is an important indicator of schistosomiasis endemic rebound. Therefore, the snail control is one of the most important schistosomiasis control measures.

[Selection of epidemic indicators for schistosomiasis GIS platform in Dongting Lake area].

Dongting Lake area is one of the major marshland schistosomiasis endemic areas in China. In recent years, spatial epidemiology is widely used in the research of schistosomiasis, which is a new opportunity to break through the current wandering situation of schistosomiasis control. In this article, both the generalized and Dongting-Lake-specific epidemic indicators of schistosomiasis are reviewed to provide the basis to construct the schistosomiasis Geographic Information System (GIS) database of Hunan Province.