Feasibility of salvage colonoscopy by water exchange for failed air-insufflated patients: a prospective, randomized, controlled trial.

BACKGROUND: A complete colonoscopy is crucial for screening colorectal diseases and colorectal cancer. However, a failure rate of up to 43% still exists. Several studies have indicated that the water exchange method can enhance the cecal intubation rate while reducing discomfort of the patient. Water exchange colonoscopy (WEC) might be a salvage treatment for the patients who failed from air insufflation colonoscopy (AIC). We aimed to assess the feasibility of WEC as a salvage measure following the failure of conventional AIC. METHODS: Patients willing to undergo unsedated colonoscopy at a tertiary-care referral center in China were randomly assigned 1:1 to WEC or AIC group for salvage after the initial AIC attempt failed. Patients were blinded to group assignment. The primary outcome was cecal intubation rate, the secondary outcomes included time to the cecum, maximum pain scores, and technical difficulty level. RESULTS: Recruited 104 patients were randomized to the WEC (n = 52) or AIC (n = 52) group. WEC significantly increased the cecal intubation rate (92.3% vs 73.1%; p = .02). The maximum pain scores and technical difficulty level in the WEC group were significantly lower than the AIC group during salvage procedure (p < .001). CONCLUSIONS: This randomized, controlled trial confirms that the WEC significantly enhanced cecal intubation rate in difficult colonoscopy in unsedated patients after the failure of standard AIC. The increased cecal intubation rate, lower pain scores and technical difficulty level suggest WEC is a good alternative for incomplete unsedated colonoscopy. Clinical trial registration number: ChiCTR2100051483.

The AC006262.5-miR-7855-5p-BPY2C axis facilitates hepatocellular carcinoma proliferation and migration.

Hepatocellular carcinoma (HCC) is typically fatal, and patients with hepatocellular carcinoma are usually diagnosed at the late stages. Although the treatments for HCC have been rapidly advancing, novel targets for HCC are still desperately needed, especially for targeted therapies. Here, we identified an enriched long non-coding RNA, AC006262.5, associated with HCC, that promoted the proliferation, migration, and invasiveness of HCC cells, both in vitro and in vivo. In addition, our results revealed that AC006262.5 bound to and regulated miR-7855-5p, a tumor-suppressive miRNA, in HCC. Moreover, our data show that AC006262.5 regulates the expression of BPY2C via miR-7855-5p. Finally, we found that AC006262.5 and miR-7855-5p formed a regulatory loop. Upregulation of AC006262.5 resulted in decreased expression of miR-7855-5p, and downregulation of miR-7855-5p further facilitated the expression of AC006262.5. Our work provides novel targets for HCC diagnosis and treatment, and sheds light on the lncRNA-miRNA regulatory nexus that controls the pathology of HCC.

[Expression of CXCR7 in Gastric Cancer Cells and Its Effect on the Migration and Invasion of SGC-7901 Cells].

OBJECTIVES: To determine the expression of chemokine (C-X-C motif) receptor 7 (CXCR7) in five gastric cancer cell lines with various degrees of differentiation, and the effect of silencing CXCR7 on the migration and invasion of SGC-7901 cells. METHODS: The expression of CXCR7 in gastric cell lines (HGC-27, MGC-803, SGC-7901, BGC-823 and MKN-28) was detected by Western bolt and RT-PCR. The SGC-7901 cells were transfected with liposome of CXCR7 siRNA to silence CXCR7 gene, and then treated with stromal-derived factor-1 (SDF-1)-the ligand of CXCR7. Transwell assay was used for determining the migratory and invasive ability of SGC-7901 cells in the four groups: NC siRNA, NC siRNA+SDF-1, CXCR7 siRNA and CXCR7 siRNA+SDF-1. RESULTS: CXCR7 was expressed in the five gastric cancer cell lines, with the highest intensity in SGC-7901. The migrated and invasive cells increased in the NC siRNA+SDF-1 group and reduced in the CXCR7-siRNA group compared with the NC siRNA group (P<0.05). The CXCR7-siRNA+SDF-1 group had less migrated and invasive cells than the NC siRNA+SDF-1 group (P<0.05). CONCLUSIONS: CXCR7 is highly expressed in SGC-7901. SDF-1 promotes the migratory and invasive capability of SGC-7901 cells, but such an effect can be inhibited by silencing it with CXCR7siRNA.

[Effect of Chlorogenic Acid on Hepatic Stellate Cell Proliferation, Generation and Degradation of Extracelluar Matrix].

OBJECTIVE: To investigate the effects of chlorogenic acid (CGA) on hepatic stellate cell proliferation and the expression and secretion of Collagen I, Collagen III, tissue inhibitors of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-2 (MMP-2). METHODS: An immortalized rat hepatic stellate cell (HSC) line was cultured in vitro. The cells were divided into 5 groups: control group; platelet-derived growth factor (PDGF) (10 ng/mL), PDGF+CGA (12.5 µg/mL), PDGF+CGA (25 µg/mL), PDGF+CGA (50 µg/mL) and CGA (50 µg/mL) group. After 24 hours treatment, the proliferation of HSC was detected by MTT method. The mRNA expression of Collagen I, Collagen III, TIMP-1 and MMP-2 were detected by RT-PCR. The protein levels of Collagen I, Collagen III, TIMP-1 and MMP-2 in the culture supernatant of HSC were measured by ELISA. RESULTS: PDGF increased the hepatic stellate cell proliferation, the mRNA expression and the protein levels of Collagen I, Collagen III and TIMP-1. (P < 0.05), which were significantly decreased by CGA (P < 0. 05). However, CGA had no significant influence on the expression of MMP-2. CONCLUSION: The antifibrotic effect of CGA may be related with the inhibition of hepatic stellate cell proliferation and generation of extracelluar matrix and promotion of extracelluar matrix degradation.

[Experimental study on effect of model on hepatic fibrosis with Aralia chinesis].

Hepatic fibrosis models were induced by CCl4 in rats. To explore vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGFß1) mRNA expression and bcl-2, Bax protein expression levels of intervention and explore the mechanism of the Aralia chinesis anti-hepatic fibrosis. Sixty male Sprague-Dawlley (SD) rats were randomly divided into six groups: nomal group, model group, high-dose (10 mL x kg(-1)), medium-dose (7.5 mL x kg(-1)), low-dose (5.0 mL x kg(-1)) of A. chinesis treated group and colchicine treated group. The change of liver histopathology was observed by HE and Masson staining. The mRNA of VEGF, TGF-ß1 were detected by RT-PCR. The protein of Bcl-2 and Bax were detected by Western blot. In the model group liver cell obvious degeneration, necrosis, a large number of collagen fibers of the cable hyperplasia, part visible pseudolobule formation. A. chinesis large, medium, low-dose group and colchicine group liver cell degeneration and necrosis reduced A. chinesis small, medium, and high-dose group was gradually reduced trend and A. chinesis large, middle dose group degree of reduction is particularly significant. Compared with model group, A. chinesis of large, medium and small dose group and colchicine group VEGF mRNA expression, A. chinesis of large, medium-dose group TGF-ß1 mRNA expression reduce (P < 0.05); compared with colchicine group, A. chinesis of large, middle dose group of VEGF mRNA expression decreased (P < 0.05); A. chinesis of large, middle dose group of TGF-ß1 mRNA expression decreased (P < 0.01), and compared with colchicine group, large dose group of of TGF-ß1 mRNA expression decreased (P < 0.05). Compared with model group, A. chinesis of large, medium and small dose group and colchicine group Bcl-2 protein expression reduce (all is P < 0.05). But A. chinesis of large, medium and small dose group and colchicine group of Bax protein expression were increased (P < 0.05). A. chinesis regulation of VEGF, TGF-ß1 may prevent the activation of hepatic stellate cells, liver tissue by up regulating the anti-apoptotic protein Bax and down pro-apoptotic protein Bcl-2 expression, thereby to improve the degree of liver fibrosis.

[Effect of alcohol extract of Plumula Nelumbini on carbon tetrachloride induced rat liver fibrosis: an experimental study].

OBJECTIVE: To study the protective effect of alcohol extract of Plumula Nelumbini (AEPN) on carbon tetrachloride (CCl4) induced hepatic fibrosis rats and to explore its possible mechanism. METHODS: Totally 32 male SD rats were randomly divided into four groups, i.e., the normal control group, the model group, the high dose AEPN group, and the low dose AEPN group, 8 in each group. 1,000 mg/kg AEPN was given to rats in the high dose AEPN group by gastrogavage at 10 mL/kg, once daily, while 500 mg/kg AEPN was given to rats in the low dose AEPN group by gastrogavage at 10 mL/kg, once daily. Hepatic fibrosis was induced by intraperitoneal injection of CCl4. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin (ALB) were examined using automatic biochemical analyzer. Activities of superoxide dismutase (SOD), contents of malondialdehyde (MDA) and hydroxyproline (Hyp) in the hepatic tissue were determined using colorimetry. The degree of liver fibrosis was observed by HE staining and Masson staining. The expression of α-smooth muscle actin (α-SMA) was detected using immunohistochemistry. RESULTS: (1) Compared with the normal control group, serum levels of ALT and AST obviously increased and the serum ALB level obviously decreased in the model group (all P < 0.05). After treated by AEPN, serum levels of ALT and AST were lowered. and the serum ALB level was higher (all P < 0.05). (2) Compared with the normal control group, collagen deposition was obviously seen in rats' livers of the model group, and pseudolobule had formed; inflammatory activities and fibrosis degrees were serious; contents of Hyp also increased (P < 0.05).After treated by AEPN, collagen deposition was obviously reduced with no obvious pseudolobule; inflammatory activities and fibrosis degrees were alleviated; contents of Hyp were also lowered (P < 0.05). (3) Compared with the normal control group, contents of MDA in the liver tissue obviously increased, while activities of SOD obviously decreased (P < 0.05) in the model group. After treated by AEPN, contents of MDA in the liver tissue decreased and the serum SOD level significantly increased (all P < 0.05). (4) Compared with the normal control group, the expression of α-SMA was obviously elevated in the model group (P < 0.05). After treated by AEPN, its expression was obviously lowered (P < 0.05). CONCLUSIONS: AEPN could fight against CCl4 induced liver fibrosis in rats. Fighting against lipid peroxidation and inhibi- ting activation and proliferation of hepatic stellate cells might be possibly main mechanism.

Accumulation rate and sources of plastic marine litter at nesting grounds of green turtles on the North Island of Qilianyu, Xisha Islands, South China Sea.

In this study, the accumulation rate of plastic litter was investigated by sampling quadrats placed on the North Island of Qilianyu, and the composition was analyzed and identified to determine its source. The results showed that the annual average accumulation rate of plastic litter on North Island was 0.64 ± 0.32 pieces·m-2·month-1, with a mass accumulation rate of 11.30 ± 7.73 g·m-2·month-1. The accumulation rate of plastic litter was mainly influenced by wind speed and direction, with higher accumulation rates occurring during the southwest monsoon season and tropical cyclones. ATR-FTIR analysis indicated that polyethylene (44 %) and polypropylene (41 %) were the most abundant types of polymers. This study reveals the current status of plastic litter pollution in green turtle nesting grounds on North Island in Qilianyu, which can be used as a reference for management strategies that mitigate plastic litter pollution.

Genealogy and Demographic History of a Widespread Amphibian throughout Indochina.

Relatively little is known about spatial patterns of cryptic diversity in tropical species and the processes that generate them. Few studies examine the geographic distribution of genetic lineages in Southeast Asia, an area hypothesized to harbor substantial cryptic diversity. We investigated the evolutionary history of Asian tree frogs of the Polypedates leucomystax complex (n = 172) based on 1800 bp of the mtDNA genes ND1 and cytochrome b and tested hypotheses pertaining to climate, geology, and dispersal patterns. Analyses revealed substantial genetic diversity and lineage divergence throughout the region with evidence for widespread sympatric lineages and a general north versus south clustering. Relaxed molecular clock analysis and tests for demographic expansion identified an initial cladogenesis during the Miocene with subsequent Plio-Pleistocene diversification, with the former corresponding to periods of increased aridity and the onset of monsoonal weather systems. Rates of diversification were relatively constant until the Early Pleistocene when rates increased exponentially. We found equivocal evidence for isolation-by-distance and a potential role of some landscape features as partial barriers to dispersal. Finally, our analyses showed that divergence between insular and mainland populations occurred before Homo sapiens colonized Southeast Asia, suggesting that historical human-mediated dispersal did not drive insular diversification. Our results suggested that demographic expansion in the Late Pleistocene resulted in widespread sympatric lineages in the P. leucomystax complex throughout southern China and Indochina and further clarified the evolutionary history of lineages within P. leucomystax.

[Effect of neferine on hepatic stellate cells in collagen-I, TIMP-1 and MMP-2].

OBJECTIVE: To observe the effect of neferine on Collagen-I, TIMP-1 and MMP-2 expressions and protein secretion of hepatic stellate cells. METHOD: The hepatic stellate cell line HSC-T6 was cultured in vitro, and then randomly divided into 5 groups: the control group, the platelet-derived growth factor (PDGF) group and PDGF + neferine (2, 6, 10 micromol x L(-1)) groups. All of the groups were cultured for 48 h, and their cells were collected to extract mRNA and detect Collagen-I, TIMP-1 and MMP-2 expressions with RT-PCR. Their cell supernatants were also collected to determine the protein content of three factors with ELISA. RESULT: Compared with the control group, PDGF could remarkably increase the Collagen-I, TIMP-1 and MMP-2 expressions and protein secretion of hepatic stellate cells. Compared with the PDGF group, PDGF + neferine (6, 10 micromol x L(-1)) groups showed a notable decrease in the Collagen-I and mRNA expression and protein secretion along with the increase in the concentration, whereas the PDGF + neferine (2 micromol x L(-1)) group showed no significant change in the Collagen-I and mRNA expression and protein secretion. Compared with the PDGF group, three PDGF + neferine groups showed no notable change in MMP-2 expression and protein secretion. CONCLUSION: Neferine can inhibit the Collagen-I, TIMP-1 and mRNA protein expression and protein secretion of PDGF-induced HSCs along with the increase in the concentration, but with not remarkable effect on the MMP-2 expression and secretion.

Microhabitat characteristics of the critically endangered big-headed turtle Platysternon megacephalum.

Understanding the microhabitat requirements of an animal is vital for ensuring the success of targeted conservation and microhabitat restoration measures. The big-headed turtle (Platysternon megacephalum) is a freshwater species that is distributed across Southeast Asia. Owing to the human threats posed by illegal pet trade and overharvesting for food and medicinal purposes, the species has undergone rapid decline. However, in Hainan, their microhabitat characteristics are still unknown, which is neither conducive to the conservation of the species nor to the establishment of the Hainan Tropical Rainforest National Park. This study examined the microhabitat characteristics of P. megacephalum using sample plot methods in the Diaoluo Mountain area of Hainan Tropical Rainforest National Park. Our results indicated that P. megacephalum prefers stream microhabitats with rocky substrates, several caves, and a high diversity of food sources. Microhabitat characteristics did not differ significantly between adults and juveniles. Our results suggest that protecting microhabitats and main food sources is important for the conservation of P. megacephalum. Our findings provide a reference for the protection of this species in Jianfeng Ridge, Yingge Ridge, and other areas in Hainan Tropical Rainforest National Park.

Nocturnal emergence facilitated by thermally-induced hatching in the Chinese softshell turtle, Pelodiscus sinensis.

The coincidence of hatching and emergence events with favorable conditions is crucial for turtle survival. Nocturnal emergence has been widely documented across marine and freshwater turtles, and has long been suggested as an adaptive behavior that reduces risks of heat stress and predation. To our knowledge, however, studies related to nocturnal emergence have mainly focused on the post-hatching behaviors of turtles, and very few experimental studies have been performed to investigate the effects of hatching time on the distribution of emergence times over the course of a day. Here, we visually monitored the activity of the Chinese softshell turtle (Pelodiscus sinensis)-a shallow-nesting freshwater turtle-from hatching to emergence. Our study provides evidence for the novel finding that (i) the timing of synchronous hatching events in P. sinensis coincides with the time of day when nest temperatures decrease, (ii) the synchrony between hatching and emergence may further facilitate their nocturnal emergence, and (iii) synchronous behaviors of hatchlings in the nest may be effective in reducing the risk of hatchling predation, and predation is more likely to occur in the asynchronous hatching groups. This study suggests that the hatching of shallow-nesting P. sinensis in response to temperature changes in the nest might be an adaptive nocturnal emergence strategy.

Investigation on beach debris on the historical nesting grounds of green turtles (Chelonia mydas) in Hainan Island, South China.

Hainan Island used to be the most important nesting ground of green turtles in China before they disappeared about 37 years ago. Habitat degradation is one of the main reasons for the disappearance of sea turtles. Therefore, it is necessary to take action to evaluate and recover the historical nesting grounds if we hope for sea turtles to return in the future. In this study, we surveyed the beach debris on 13 historical nesting grounds of green sea turtles on Hainan Island. The beach debris on these nesting grounds mainly consisted of plastic, cigarette butts, foam, glass, and nylon, with plastic (including plastic blocks, cigarette butts, and foam) being the dominant type, accounting for 78.92% in number, followed by glass. The average density of beach debris was 0.314 pieces·m-2. Compared to other nesting grounds, the average quantity and density of beach debris in Hainan was lower, but the proportion of plastic debris was extremely high. After categorizing debris type, we found that most was from human coastal activities (35.54%), with debris at tourist beaches having the biggest proportion of debris from smoking supplies. The distribution characteristics of beach debris were related to the function of the beach, density of tourist, and the intensity of beach debris cleaning. It is recommended to further strengthen the emission reduction and clean-up of beach debris in Hainan Island, so as to restore the nesting habitat of sea turtles as soon as possible.

Revisiting the genetic diversity and population structure of the endangered Green Sea Turtle (Chelonia mydas) breeding populations in the Xisha (Paracel) Islands, South China Sea.

The Green Sea Turtle (Chelonia mydas) is an umbrella species in the South China Sea, a Chinese national first-level protected wild animal, and the only sea turtle that nests in waters around China. The largest C. mydas nesting ground is distributed in the Xisha (Paracel) Islands, which plays a vital role in the survival of sea turtle populations in the region. This study reveals the genetic diversity and population structure of the breeding population of C. mydas in the Xisha (Paracel) Islands using three mitochondrial markers. A total of 15 D-loop, five Cytochrome b (Cyt b), and seven Cytochrome C Oxidase subunit I (COI) haplotypes were identified in the breeding population of C. mydas in the Xisha (Paracel) Islands. D-loop haplotypes are distributed in clades III, IV, and VIII of the C. mydas mitochondrial control region. It is the first time that one haplotype from Clade IV was found in this C. mydas population, and five new D-loop haplotypes were also identified. The haplotype and nucleotide diversity were calculated for each marker: D-loop (0.415 haplotype diversity, 0.00204 nucleotide diversity), Cyt b (0.140, 0.00038) and COI (0.308, 0.00083). The average genetic distance (p) of each molecular marker was less than 0.01. Neutral detection and nucleotide mismatch analysis suggested that the breeding population of C. mydas in the Xisha (Paracel) Islands did not experience a population expansion event in recent history. It is recommended that a sea turtle protection area be established in the Xisha (Paracel) Islands area to strengthen protection and effectively protect the uniqueness and sustainability of the breeding population of C. mydas in the South China Sea.

Increasing nitric oxide content in Arabidopsis thaliana by expressing rat neuronal nitric oxide synthase resulted in enhanced stress tolerance.

Nitric oxide (NO) plays essential roles in many physiological and developmental processes in plants, including biotic and abiotic stresses, which have adverse effects on agricultural production. However, due to the lack of findings regarding nitric oxide synthase (NOS), many difficulties arise in investigating the physiological roles of NO in vivo and thus its utilization for genetic engineering. Here, to explore the possibility of manipulating the endogenous NO level, rat neuronal NOS (nNOS) was expressed in Arabidopsis thaliana. The 35S::nNOS plants showed higher NOS activity and accumulation of NO using the fluorescent probe 3-amino, 4-aminomethyl-2', 7'-difluorescein, diacetate (DAF-FM DA) assay and the hemoglobin assay. Compared with the wild type, the 35S::nNOS plants displayed improved salt and drought tolerance, which was further confirmed by changes in physiological parameters including reduced water loss rate, reduced stomatal aperture, and altered proline and malondialdehyde content. Quantitative real-time PCR analyses revealed that the expression of several stress-regulated genes was up-regulated in the transgenic lines. Furthermore, the transgenic lines also showed enhanced disease resistance against Pseudomonas syringae pv. tomato (Pst) DC3000 by activating the expression of defense-related genes. In addition, we found that the 35S::nNOS lines flowered late by regulating the expression of CO, FLC and LFY genes. Together, these results demonstrated that it is a useful strategy to exploit the roles of plant NO in various processes by the expression of rat nNOS. The approach may also be useful for genetic engineering of crops with increased environmental adaptations.

The expression of protease-activated receptor 2 and 4 in the colon of irritable bowel syndrome patients.

BACKGROUND: Irritable bowel syndrome (IBS) pathophysiology has not been fully understood. Abnormalities of serine proteases have been identified in IBS patients. In addition, protease-activated receptors (PAR) activation interferes with several components of the pathogenesis of IBS, so, evaluating the PAR expression in IBS patients may contribute to understanding the pathogenesis of the disease. AIMS: This study aimed to investigate whether the expression of PAR4 and PAR2 in the colon was changed in IBS patients and was associated with IBS. METHODS: Colon mucosal biopsies of 34 IBS patients (16 constipation- and 18 diarrhea-predominant) and 18 control subjects were collected. Gene transcripts of PAR2, PAR4, tryptase, and trypsin were quantified using real-time polymerase chain reaction (PCR) and the expression of PAR2 and PAR4 receptors was also measured by immunohistology and image analysis. RESULTS: In IBS patients, the mRNA expression of tryptase and trypsin normalized against ß-actin gene was higher compared to control subjects (P < 0.001). No difference was observed in the PAR2 mRNA level or protein level between control subjects on the one hand and IBS patients or subgroups on the other. In IBS or IBS subgroups patients, the expression of PAR4 in the mRNA level or protein level was lower than the control subjects. CONCLUSIONS: This study, for the first time, indicated the PAR4 expression in IBS patients. Decreased PAR4 expression may help us to understand the pathogenesis of IBS.

[Effects of Busui Shengxue Granule on bFGF and bFGFR in bone marrow derived stroma cells of chronic aplastic anemia patients].

OBJECTIVE: To observe the therapeutic effects of Busui Shengxue Granule (BSSXG) on chronic aplastic anemia (CAA) patients and its effects on bone marrow derived stroma cells (BMDSCs) correlated cytokines. METHODS: One hundred and twenty-four patients with CAA were randomly assigned to two groups according to the random digit table. Patients in the test group (61 cases) were treated with BSSXG, while those in the control group (63 cases) were treated with Zaizao Shengxue Tablet (ZST). The therapeutic course was 6 months for all. Besides, 10 healthy subjects were recruited as the normal control group. Changes of the symptom integral, therapeutic efficacy judgment, and changes of peripheral hemogram of patients were observed. The mRNA expression of b-fibroblast growth factors (bFGF) and b-fibroblast growth factors receptor (bFGFR) were detected by reverse transcription PCR. RESULTS: The total effective rate of the test group was 75.0% (45/61), higher than that of the control group (58.7%, 37/63). Its symptom integral and peripheral hemogram were obviously improved, better than those of the control group (P < 0.05, P < 0.01). The mRNA expressions of bFGF and bFGFR of the test group were obviously lower than those of the normal control group (P < 0.05, P < 0.01). They were somewhat improved after treatment in the two groups, with better results obtained in the test group. CONCLUSIONS: BSSXG showed better clinical effects. It could improve the symptom integral and peripheral hemogram of CAA patients, improve the clinical efficacy, and regulate the expression levels of bFGF and bFGFR. It improved the hematopoietic microenvironment and promoted the hematopoiesis of the bone marrow through regulating the proliferation and oriental differentiation of stroma cells, and promoting the bone marrow angiogenesis.

[Changes in the intestinal microenvironment during development of alcoholic fatty liver disease and related effects of probiotic therapy].

OBJECTIVE: To investigate the initial changes in the gut microenvironment that accompany intestinal endotoxemia related to alcoholic fatty liver disease (ALD) in order to explore the potential initiating factors and to observe the effect of probiotic therapy on these factors. METHODS: Fifty Sprague-Dawley male rats were randomly divided into an ALD model group (alcoholic intragastric administration), an intervention group (ALD with probiotic intragastric administration), and a control group (physiological saline intragastric administration). Histological changes of the liver were evaluated using hematoxylin-eosin staining and light microscopy. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides (TG), and plasma endotoxin and coli bacillus were determined. The structural integrity of intestinal mucosa and tight junctions were observed by transmission electron microscopy. Occludin protein expression in intestinal epithelial cells was detected by immunohistochemistry. RESULTS: After four weeks, the three groups showed significant differences in the plasma endotoxin levels [control: (0.67+/-0.14) pg/ml, model: (4.42+/-1.28) pg/ml, and intervention: (2.88+/-0.83) pg/ml; F = 27.288, P = 0.000] and numbers of Escherichia coli [control: (2.31+/-0.39) lg3/ml, model: (3.23+/-0.41) lg3/ml, and intervention: (2.24+/-0.44) lg3/ml; F = 10.692, P = 0.001]. The plasma endotoxin level and E. coli number were significantly higher in the model group than in the control group and the intervention group (all P less than 0.05). The three groups showed no significant differences in the levels of ALT, AST, and TG at four weeks. After eight weeks, however, all three serum markers were significantly different between the three groups [ALT: control: (62.33+/-7.12) U/L, model: (95.50+/-8.73) U/L, and intervention: (81.33+/-6.19) U/L; F = 18.051, P = 0.000]; [AST: control: (90.50+/-10.67) U/L, model: (130.00+/-14.91) U/L, and intervention: (110.33+/-7.26) U/L; F = 30.170, P = 0.000]; [TG: control: (0.84+/-0.84) mmol/L, model: (1.40+/-0.17) mmol/L, and intervention: (1.10+/-0.17) mmol/L; F = 10.592, P = 0.001]. In addition, the three groups showed significant differences in E. coli number [control: (2.23+/-0.46) lg3/ml, model: (4.81+/-0.29) lg3/ml, and intervention: (3.61+/-0.50) lg3/ml; F = 23.579, P = 0.000] and plasma endotoxin level [control: (0.52+/-0.21) pg/ml, model: (12.46+/-2.61) pg/ml, intervention: (6.83+/-1.74) pg/ml; F = 30.731, P = 0.000]. The levels of ALT, AST, TG and endotoxin, and the number of E. coli were all significantly higher in the model group than in the control group and the intervention group (all P less than 0.05). Small intestinal epithelial cell structural failure was more apparent and intercellular gaps more broad after eight weeks than after four weeks for all three groups. However, the intervention group showed clearer cell connection structures and less extensive cell gap broadening than the model group at eight weeks. After eight weeks, the occludin protein had become significantly down-regulated and distributed in a non-continuous pattern in the model group, as compared with the control group. However, the occludin protein expression was higher in intervention group than in the model group. CONCLUSION: Intestinal endotoxemia related to perturbations in the microenvironment occurs in the early phase of ALD, and the increased intestinal permeability appears to be the initial factor of elevated plasma endotoxin, which may lead to liver damage. Probiotic therapy can reduced plasma endotoxin levels and postpone ALD progression by altering the composition of the gut microbiota and up-regulating expression of the occludin protein in intestinal epithelial cells.

Phylogeography of the endangered "eyed" turtles (genus Sacalia) and the discovery of a lineage derived from natural interspecific hybridization.

The herpetofauna of the Indomalayan bioregion of Asia suffers from severe habitat loss, unsustainable harvesting, and lack of research and conservation. Here, we investigated the range-wide phylogeography of the endangered "eyed" turtles (genus Sacalia, including the Beale's Eyed Turtle S. bealei and the Four-eyed Turtle S. quadriocellata) and discovered a natural interspecific hybrid turtle population in China. Based on phylogeny of the mitochondrial Cytochrome b gene of 101 samples in this study and public data, three major clades and six subclades were identified: S. bealei (SBE) in eastern-southern China, east S. quadriocellata in South China (northern east [SQUen] and southern east [SQUes] subclades), and west S. quadriocellata mainly in Vietnam (northern west [SQUwn], central west [SQUwc], and southern west [SQUws] subclades). We sequenced 16 nuclear DNA loci of 87 samples from SBE, SQUen, SQUes, and SQUwn subclades. Population genetic clustering analysis suggested a structure similar to the mitochondrial phylogeny, where most samples were classified into four genetic clusters corresponding to the four mtDNA subclades. However, a proportion of samples carrying SQUen mtDNA haplotypes formed an additional distinct cluster SHY. Those samples are found in the contact zone of the two species bearing mosaic and intermediate morphological characteristics. We detected an admixed ancestry in SHY from SBE and SQUen that conformed to an intrapopulation breeding scenario for at least hundreds of generations after the initial hybrid event, leading to a conclusion that SHY is a distinct and near-panmictic population derived from natural interspecific hybridization. In addition, SQUes (Hainan Island endemic) is of special concern due to significant isolation and low genetic diversity. We suggest that seven evolutionarily significant units should be recognized to facilitate appropriate conservation actions. These findings also highlight the urgent need for further herpetological research and conservation in this region.