Search

BACKGROUND Sulongga-4 (SL-4) is a herbal formula used in traditional Mongolian medical clinics for the treatment of peptic ulcers and gastroenteritis, even though its pharmacological mechanism has not been well characterized. AIM To evaluate the protective effect and identify the mechanisms of action of SL-4 on gastroduodenal ulcer induced by pyloric ligation (PL) in rats. METHODS PL was performed to induce gastric and duodenal ulcers in rats, which were then treated with oral SL-4 (1.3, 2.6, or 3.9 g/kg per day) for 15 d. PL-induced gastroduodenal ulceration. Therapeutic effects were characterized by pathological and histological evaluations and inflammatory indicators were analyzed by enzyme-linked immunosorbent assay. Microarray analyses were conducted to identify gene expression profiles of gastroduodenal tissue in PL rats with or without SL-4 treatment. The candidate target genes were selected and verified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS SL-4 decreased histopathological features in the PL-induced ulcerated rats. SL-4 significantly (P < 0.05) decreased expression of tumor necrosis factor-α, interleukin (IL)-1β, IL-6, endotoxin, platelet-activating factor, and increased prostaglandin E2 and epidermal growth factor in ulcer tissue. Microarray analysis was used to identify a panel of candidate target genes for SL-4 acting on PL-induced ulceration. Genes included some complement and coagulation cascade and retinol metabolism pathways that are closely associated with inflammatory responses and gastric mucosal protective mechanisms. qRT-PCR showed that altered expression of the selected genes, such as CYP2b2, UGT2b1, A2m, and MASP1 was consistent with the microarray results. CONCLUSION SL-4 exerts protective effects against PL-induced gastroduodenal ulcers via reducing inflammatory cytokines and elevating expression of gastric acid inhibitory factors. Downregulation of CYP2b2 and UGT2b1 genes in retinol metabolism and upregulation of A2m and MASP1 genes in the complement and coagulation cascades pathways are possibly involved in SL-4-mediated protection against gastroduodenal ulcer.

Background & aims Insulin resistance is strongly associated with non-alcoholic fatty liver disease, a chronic, obesity-related liver disease. Increased endoplasmic reticulum (ER) stress plays an important role in the development of insulin resistance. In this study, we investigated the roles of miRNAs in regulating ER stress in the liver of rats with obesity. Methods We used miRNA microarray to determine the miRNA expression profiles in the liver of rats fed with a high fat diet (HFD). We used prediction algorithms and luciferase reporter assay to identify the target gene of miRNAs. To overexpress the miRNA miR-30b or inhibit miR-30b rats were injected with lentivirus particles containing PGLV3-miR-30b or PGLV3-miR-30b antimiR through tail vein. Hepatic steatosis was measured using transient elastography in human subjects. Results Our data showed that miR-30b was markedly up-regulated in the liver of HFD-treated rats. Bioinformatic and in vitro and in vivo studies led us to identify sarco(endo)plasmic reticulum Ca2+ -ATPase 2b (SERCA2b), as a novel target of miR-30b. Overexpression of miR-30b induced ER stress and insulin resistance in rats fed with normal diet, whereas inhibition of miR-30b by miR-30b antimiR suppressed ER stress and insulin resistance in HFD-treated rats. Finally, our data demonstrated that there was a positive correlation between serum miR-30b levels and hepatic steatosis or homoeostasis model assessment of insulin resistance (HOMA-IR) in human subjects. Conclusions Our findings suggest that miR-30b represents not only a potential target for the treatment of insulin resistance, but also a non-invasive disease biomarker of NAFLD.

Contaminating organisms in mass cultivation present one of the major challenges that must be overcome for successful commercialization of algal biofuels. The present study identified a range of contaminating organisms in Chlorella cultures cultivated in outdoor raceway ponds at the Arizona Center for Algae Technology and Innovation (Mesa, AZ). Nineteen organisms or Operational Taxonomic Units (OTU) in the Chlorella culture were identified by a combination of microscopic observation and 18S rRNA denaturing gradient gel electrophoresis (DGGE). More detailed analyses identified these contaminating organisms as 2 fungi, 7 flagellates, 3 amoebae, 4 ciliates, 1 rotifer, and 2 large insects. Among them Poterioochromonas sp., a small flagellate, appeared to be one of the most harmful causing culture collapse. In order to control Poterioochromonas sp., various operational parameters and application strategies of ultrasonic treatment were investigated. During sixteen-day consecutive Chlorella cultivation in batch mode, the ultrasonication conditions of 6 L min−1 flow rate with the power of 495 W at 100% amplitude and a treatment frequency of once for 1 h every day, was proven to be the most effective in preventing Poterioochromonas outbreak in Chlorella culture with volume of 60 L. The above ultra-sonication method was also effective at destroying an unknown fungus, an amoeba (Acanthocystis sp.), and ciliates (a member of the family Orchitophryidae). Our findings can serve as a technical foundation for the application of ultrasonication to control some of the contaminating microorganisms in mass cultivation of microalgae including Chlorella.

Pathogen avoidance behaviors are found throughout the animal kingdom and are important for animal's survival in nature. As a free-living nematode, C. elegans is exposed to a variety of microorganisms, including toxic or pathogenic bacteria, in soil. C. elegans can develop efficient avoidance responses to pathogenic bacteria to minimize the infection risk. However, the role of microRNAs (miRNAs) in pathogen avoidance in C. elegans remains unclear. In this report, we showed that the miRNA mir-67 was involved in a behavioral avoidance response to P. aeruginosa PA14. Exposure to P. aeruginosa PA14 induced the expression of mir-67 in worms. mir-67(n4899) mutants exhibited a reduced ability to avoid P. aeruginosa PA14. By combining quantitative proteomic analysis with miRNA target prediction algorithms, we identified SAX-7/L1CAM, which is transmembrane cell adhesion receptor molecule, as the target of mir-67. Silencing of sax-7 by RNAi on mir-67 mutants rescued avoidance behavioral. Our data demonstrate that the mir-67-SAX-7 pathway modulate the behavioral avoidance response to pathogens, thus providing a new perspective in the role of miRNAs in host-microbe interactions.

We study numerically the propagation of Lamb waves in one-dimensional (1D) quasi-periodic composite thin plates consisting of a row of air holes embedded in the matrix material silicon according to a Cantor sequence; the surfaces of the plate are parallel to the axis of quasi-periodicity. The phenomenon of multi-splitting in the band gap structures is demonstrated. A semi-quantitative explanation is proposed in which the inherent cavity-like structure is proven to play the essential role in the phenomenon of multi-splitting, which gives a reliable way to predict where and how the band gap is splitting in the quasi-periodic systems. Possible applications are discussed.

In many wireless sensor network applications, it should be considered that how to trade off the inherent conflict between energy efficient communication and desired quality of service such as real-time and reliability of transportation. In this paper, a novel routing protocols named balance energy-efficient and real-time with reliable communication (BERR) for wireless sensor networks (WSNs) are proposed, which considers the joint performances of real-time, energy efficiency and reliability. In BERR, a node, which is preparing to transmit data packets to sink node, estimates the energy cost, hop count value to sink node and reliability using local information gained from neighbor nodes. BERR considers not only each sender' energy level but also that of its neighbor nodes, so that the better energy conditions a node has, the more probability it will be to be chosen as the next relay node. To enhance real-time delivery, it will choose the node with smaller hop count value to sink node as the possible relay candidate. To improve reliability, it adopts retransmission mechanism. Simulation results show that BERR has better performances in term of energy consumption, network lifetime, reliability and small transmitting delay.

40t semi-trailer frame is optimized by usage of topological optimization and static/dynamic analysis technology. After the optimization, The static characteristics of frame completely satisfy requirement of intensity and stiffness, and the quality is reduced by 10% compared with the original frame. The dynamic characteristics of the new frame have been improved greatly, which indicate that the optimized frame structure is more reasonable, and lay the foundation for reducing the cost and improving manufacturing efficiency.

In the paper, the emulsion graft polymerization is used to prepare stable epoxy resin emulsion by vinyl acetate and polyvinyl alcohol (PVA) as emulsification stabilizer. It is studied for the influence on the properties of emulsion by temperature and dosages of functional monomer, acrylamide and acrylic acid, and initiator. The results show that the optimal conditions: reactive temperature is 75°C, acrylic acid is 2%~4%(mass concentration), acrylamide is 2%~4%(mass concentration) and initiator is 0.7%~0.8%(mass concentration).

The research is conducted on the tank of semi-trailers which is made of steel by using the method of Finite Element Analysis. The optimized result indicated that the mass of the tank structure reduced by 9% on the premise that the intensity and the rigidity request of structural property is satisfied, and the dynamic characteristic also meets the requirement. It is the foundation to reduce the cost of manufacture and improve the operational effectiveness of user.

Taking an aluminum tank semitrailer as an example, this essay aim to analyze the strength and stiffness of the tank semitrailer and obtain stress and strain characteristics under different conditions on the basis of a finite element model, by using the finite element analysis software. By carrying on modal analysis in different work conditions, the essay gets the dynamic performance parameters of the tank. The results show that the tank fully meets the requirements of using, and the performance parameters of the tank will be foundation for the optimization.

SD rats were randomly divided into normal control, model, deep EA, and shallow EA groups. The model was established by mechanical clamping of the sciatic nerve stem. For deep and shallow EA, the needles were inserted into “Huantiao” (GB 30) by about 16 mm and 7 mm, respectively, once daily for 14 days. The results showed that, compared with the normal control group, the nerve-muscle excitability of rat’s hip muscle decreased and the nerve conduction velocity of sciatic nerve slowed down in the model group; meanwhile, the number of apoptotic cells and the expression level of Bax protein in the injured nerve increased significantly, and the expression level of Bcl-2 protein and the ratio of Bcl-2/Bax decreased considerably. Compared with the model group, the indices mentioned above were reversed in the two treatment groups, and the changes in the deep EA group were more significant than those in the shallow EA group. These results indicate that EA stimulation at GB 30 can improve the function of injured sciatic nerve, which is closely associated with its effects in upregulating the expression of apoptosis inhibitive factor Bcl-2 and downregulating apoptosis promotive factor Bax. Deep EA is relatively better.

The unfolded protein response (UPR), which is activated by perturbations of the endoplasmic reticulum homeostasis, has been shown to play an important role in innate immunity and inflammation. However, little is known about the molecular mechanisms underlying activation of the UPR during immune responses. Using small RNA deep sequencing and reverse genetic analysis, we show that the microRNA mir-233 is required for activation of the UPR in Caenorhabditis elegans exposed to Pseudomonas aeruginosa PA14. P. aeruginosa infection up-regulates the expression of mir-233 in a p38 MAPK-dependent manner. Quantitative proteomic analysis identifies SCA-1, a C. elegans homologue of the sarco/endoplasmic reticulum Ca2+-ATPase, as a target of mir-233. During P. aeruginosa PA14 infection, mir-233 represses the protein levels of SCA-1, which in turn leads to activation of the UPR. Whereas mir-233 mutants are more sensitive to P. aeruginosa infection, knockdown of sca-1 leads to enhanced resistance to the killing by P. aeruginosa. Our study indicates that microRNA-dependent pathways may have an impact on innate immunity by activating the UPR., Author Summary In the model organism Caenorhabditis elegans, the IRE1–XBP1 pathway, a major branch of the unfolded protein response (UPR), is required for host defense against pathogens. However, how innate immune responses activate the UPR is not fully understood. In this report, we find that Pseudomonas aeruginosa PA14 infection up-regulates the expression of the microRNA mir-233 in C. elegans. The response of mir-233 to P. aeruginosa PA14 infection is dependent on a major pathway of innate immunity, the p38 MAPK signaling cascade. The up-regulation of mir-233 is functionally important since a mutation in mir-233 leads to hypersensitivity of the nematode to the killing by P. aeruginosa PA14. Furthermore, we demonstrate that mir-233 contributes to the activation of the UPR by repressing the protein levels of its target SCA-1, a C. elegans homologue of the sarco/endoplasmic reticulum Ca2+-ATPase. Thus, mir-233 is an important regulator of the UPR during the innate immune response.

Autophagy, a conserved pathway that delivers intracellular materials into lysosomes for degradation, is involved in development, aging, and a variety of diseases. Accumulating evidence demonstrates that autophagy plays a protective role against infectious diseases by diminishing intracellular pathogens, including bacteria, viruses, and parasites. However, the mechanism by which autophagy regulates innate immunity remains largely unknown. Here, we show that autophagy is involved in host defense against a pathogenic bacterium Pseudomonas aeruginosa in the metazoan Caenorhabditis elegans. P. aeruginosa infection induces autophagy via a conserved extracellular signal-regulated kinase (ERK). Intriguingly, impairment of autophagy does not influence the intestinal accumulation of P. aeruginosa, but instead induces intestinal necrosis. Inhibition of necrosis results in the survival of autophagy-deficient worms after P. aeruginosa infection. These findings reveal a previously unidentified role for autophagy in protection against necrosis triggered by pathogenic bacteria in C. elegans and implicate that such a function of autophagy may be conserved through the inflammatory response in diverse organisms.

OBJECTIVE; To observe the effect of deep electroacupuncture (EA) stimulation of "Huantiao"(GB 30) on the functional and pathological changes and nerve growth factor (NGF) expression of the damaged sciatic nerve in rats, so as to study its mechanisms underlying reliving sciatica.Forty-eight SD rats were randomly divided into normal, model, deep EA and shallow EA groups (n = 12 in each group). The sciatic nerve injury model was established by mechanical clamp of the sciatic nerve stem. For deep and shallow EA, the acupuncture needles were inserted into GB 30 about 16 mm and 7 mm, respectively. The EA treatment was given 20 min, once daily for 14 days. The evoked potentials of the injured sciatic nerve stem responding to electrical stimulation were recorded by using a biophysiological experimental system for calculating the motor conduction velocity. Pathological changes of the sciatic nerve were displayed by H. E. stain. The expression of NGF and Fos proteins was detected by immunohistochemistry.In comparison with the normal group, the conduction velocity and the amplitude of the evoked potentials of the sciatic nerve were significantly decreased in the model group (P0.05). Following EA, both conduction velocity and amplitude of the evoked potentials in the deep EA group, and the conduction velocity in the shallow EA group were considerably increased (P0.05). The therapeutic effects were significantly better in the deep EA group than that in the shallow EA group (P0.05). No significant differences were found between the shallow EA and model groups in the amplitude of evoked potentials (P0.05), and no significant changes of latencies of the evoked potentials inthe four groups (P0.05). In the model group, the disorganized nerve fibers axons, myelin and Schwann cells of the damaged sciatic nerve were found, which became milder in the EA groups particularly in the deep EA group. In regard to the NGF and Fos immunoactivity of the injured sciatic nerve, the expression levels of both NGF and Fos proteins were obviously higher in the model group than in the normal group (P0.05). After EA stimulation, NGF expression was further significantly up-regulated in both deep and shallow EA groups (P0.05), and Fos protein expression was notably down-regulated in the deep and shallow EA groups (P0.05). The expression of NGF was significantly higher and Fos protein was obviously lower in the deep EA group than in the shallow EA group (P0.05).Deep EA stimulation of GB 30 can improve the pathological changes and function of the injured sciatic nerve in the rat, which is closely associated with its effects in up-regulating NGF expression and down-regulating Fos expression. The deep EA is relatively better in the therapeutic effect. These facts may be one of the mechanisms of EA in relieving sciatica in clinic practice.

In this study, researchers investigated the demographic and clinical characteristics of AIDS patients who died in hospitals, analyzed the specific causes of death, and looked for the correlation between specific cause of death and their clinical characteristics.Data of clinical characteristics of patients and their specific causes AIDS of death who died in the seven hospitals from 2009 to 2010 were collected retrospectively. All the specific causes of death were classified according to the Cause of Death (CoDe) project protocol. Univariate analysis and multivariate logistic regression analysis were used to find the association between some categorical variables and the risk for AIDS patients died from AIDS related illnesses.Clinical characteristics and the cause of death of the 381 deceased in seven hospitals in this study were collected. 82.4% were male, with priority as 30-45 years old. 123 (32.3%) death patients had received ART before death. In all death cases, the cause of death of 252 patients (66.1%) were due to AIDS related diseases, with opportunistic infections the most (92.4%). Tubercle bacillus, infection of Penicillium marneffei and Pneumocystis jiroveci were the three leading causes of opportunistic infection deaths. Of 129 patients who died of non-AIDS related disease, non-AIDS infection (29.5%), hepatitis (22.5%), and non-AIDS malignancy(10.1%)were the first three causes of death. The cause of death in patients who had injecting drug use behavior within one year, had not received ART or not long enough, with opportunistic infections, without hepatitis, with the last low CD4 cell counts before death etc. were tend to due to AIDS related disease.Opportunistic infections, non-AIDS related infections and hepatitis were the three leading causes of death in this study. The duration of time on ART had impact on the patient's cause of death. The HIV infected patients who had received ART before death had more risk to die of non-AIDS related disease, compared to patients who had not. The longer time they had accessed to ART, the less likely they would die on non-AIDS related illnesses.

To observe the CpG island methylation status, mRNA and protein expression of the Wnt inhibitory factor-1 (Wif-1) gene with procaine or 5'-Aza-2'-deoxycycytidine (5-aza-dc) on HepG2. And to explore the comparison of the demethylation with 5-aza-dc or procaine.HepG2 cells were treated with 5-aza-dc or procaine. Wif-1 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Wif-1 protein expression was detected by Western blot. The promoter methylation status of Wif-1 gene on treatment or not were detected by methylation-specific PCR (MSP).(1) RT-PCR and Western blot exhibited that both Wif-1 mRNA and Wif-1 protein expression were positive in groups treated with procaine and 5-aza-dc and highly positive in the L0 cells group, but weakly even negative in the HepG2 cells without any treatment, the difference were statistically significant (P0.05) (procaine experimental group was higher than that of 5-aza-dc experimental group, P0.05). (2) The positive rates of the promoter hypermethylated in procaine and 5-aza-de groups were (14.41 +/- 3.37)% and (29.29 +/- 1.84)%. Both of the two groups showed a part of the de-methylation status (P0.05).Both of procaine and 5-aza-dc can reverse the abnormal methylation of Wif-1 gene. Procaine can be more effective than conventional used 5-aza-dc and could be a new demethylation drug.

Liver injury is one of the most important adverse effects of antiretroviral therapy, leading to therapy changing or discontinuation. Data on liver injury in human immunodeficiency virus-1-infected patients receiving antiretroviral therapy are limited in China. The purpose of this study was to investigate the features of liver injury in human immunodeficiency virus type 1-infected patients receiving non-nucleosides reverse transcriptase inhibitors-based antiretroviral therapy in China.Seventy-five patients on antiretroviral therapy containing non-nucleosides reverse transcriptase inhibitors were retrospectively studied. The patients were divided into 2 groups: group 1 (with liver injury, n = 45) and group 2 (without liver injury, n = 30). The features of liver injury were analyzed. The sex, age, baseline CD4 counts, hepatitis B virus (HBV) and/or hepatitis C virus (HCV) co-infection, hepatotoxic drug use and nevirapine or efavirenz use were compared between two groups.Forty-five patients (60.0%), 31 (68.9%) males and 14 (31.1%) females, aged 12 to 52 years (averaged (39 ± 9) years), experienced at least one episode of liver injury. Forty (53.3%) patients were co-infected with HBV and/or HCV, 42 (56%) patients had concomitant use of antituberculosis drugs or cotrimoxazole, 46 (61.3%) and 29 (38.7%) patients received regimen containing nevirapine and efavirenz, respectively. Grade 1 liver injuries were observed in 26 (57.8%) patients, grade 2 in 16 (35.6%), grade 3 in 2 (4.0%) and grade 4 in 1 (2.2%). Three (6.7%) patients discontinued highly active antiretroviral therapy (HAART) due to liver injury. In group 1, there were 29 (64.4%) patients co-infected with HBV and/or HCV, 32 (71.1%) patients received regimen containing nevirapine, and 30 (66.7%) patients had concomitant use of anti-tuberculosis drugs or cotrimoxazole, respectively, significantly higher than those in group 2 (11 (36.7%), 14 (46.7%) and 12 (40%), respectively; P = 0.018, 0.033, 0.023, respectively). The sex, age, baseline CD4 counts and disease stage were not factors associated with liver injury.Liver injury associated with HAART containing non-nucleosides reverse transcriptase inhibitors was mild to moderate and those who were co-infected with HBV and/or HCV, had concomitant use of antituberculosis drugs or cotrimoxazole and received a regimen containing nevirapine were prone to liver injury while receiving HAART.

To comprehend the latest HIV-I epidemic tendency and the character of V3 loop in MSM population of Beijing. METHODS; The C2-V3 regions of the HIV envelop gene were amplified by nest-PCR and sequenced from 11 HIV-l-infected MSM in Beijing in 2007. The subtype and sequences of V3 loop was analyzed. RESULTS There are 4 subtype B strains, 5 CRF AE, 1 CRFO7BC and 1 CRF15-01B strains within all 11 strains. There are five types of central motifs of the 11 samples, in which GPGR and GPGQ are most common.Recombination subtype of HIV-1 are spread extensively in MSM population of Beijing.

The aim of this study was to investigate the relationship between anti-fibrotic effect of Panax notoginseng saponins (PNS) and serum cytokines in rat hepatic fibrosis. Hepatic fibrosis induced by carbon tetrachloride (CCl(4)) was studied in animal models using SD rats. Liver index, serum alanine amino transferase (ALT), aspartate amino transferase (AST), transforming growth factor-beta1 (TGF-beta1), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured, respectively. Liver index and the degree of liver fibrosis were also determined. Our results showed that the levels of ALT, AST and liver index in PNS-treated group were markedly lower than those in model group. PNS therapy also significantly attenuated the degree of hepatic fibrosis, collagen area and collagen area percent in liver tissue. Furthermore, the levels of serum TGF-beta1, TNF-alpha and IL-6 were strikingly reduced in PNS-treated group compared with model group while the production of IL-10 was up-regulated. These findings demonstrate that PNS has certain therapeutic effects on hepatic fibrosis probably by immunoregulating the imbalance between pro-fibrotic and anti-fibrotic cytokines.

To investigate the inhibitory effect of danshensu on the activation of JNK signaling in rat hepatic stellate cells (HSCs) induced by IL-1beta.The proliferation of primary rat HSCs treated with different concentration of Danshensu was checked by MTT colorimetric assay. The expression and phosphorylation of JNK and P-JNK was detected by western blotting. Synthesis and secretion of collagen I were detected by the quantitative immunocytochemical assay and ELISA.Danshensu inhibited the proliferation of HSCs in a dose-dependent manner. At the concentration of 0.0625 to 0.25 mmol/L, Danshensu significantly repressed the proliferation of HSC induced by IL-1beta (P0.05). Synthesis and secretion of Type I collagen was significantly decreased 24 hours after 0.25 mmol/L Danshensu treatment (P0.01). The phosphorylation of JNK induced by IL-1beta was significantly inhibited by Danshensu treatment (P0.05), however, the expression of JNK was not regulated by Danshensu.Danshensu represses the activation and proliferation of HSCs, and inhibits the synthesis and secretion of Type I collagen, possibly via the repression of the JNK signal transduction.

The autosomal dominant form of retinitis pigmentosa (ADRP) can be caused by mutations in 14 genes and further loci remains to be identified. This study was intended to identify mutations in a Chinese pedigree with ADRP.A large Chinese family with retinitis pigmentosa was collected. The genetic analysis of the family suggested an autosomal dominant pattern. Microsatellite (STR) markers tightly linked to genes known to be responsible for ADRP were selected for linkage analysis. Exons along with adjacent splice junctions of PRPF31 were amplified by polymerase chain reaction (PCR) and screened by direct sequencing.The caused gene of ADRP was mapped to 19q13.4 between markers D19S572 and D19S877, with a maximum LOD score of 3.01 at marker D19S418 (recombination fraction = 0).The affected gene linked to the 19q13.4 in a Chinese family with ADRP, which is different from other mutations at the same loci in other Chinese families.

To investigate the inhibitory effect of danshensu on c-Jun N-terminal kinase (JNK) and nuclear factor-kappa B (NF-kappaB) signaling in activated rat hepatic stellate cells (HSCs) and explore the mechanisms of danshensu for inhibiting hepatic fibrosis.MTT colorimetric assay was used to detect the proliferation of rat HSCs treated with danshensu, and the apoptosis of the cells was analyzed with Annexin- V-FITC/PI and AO/EB staining. The expressions of P-IkappaB-alpha, NF-kappaBP65 and JNK in HSCs stimulated by IL-1beta with subsequent danshensu treatment were observed by Western blotting. Type III collagen in the medium of HSCs was detected by ELISA and immunocytochemistry.Danshensu inhibited the activation and proliferation of HSCs, and increased the apoptotic rate of HSCs and reduced the synthesis and secretion of type III collagen. Danshensu showed obvious inhibitory effect on JNK and P-IkappaB-alpha phosphorylation and NF-kappaBP65 expression in HSCs stimulated by IL-1beta. The mechanism of the actions of dansgensu may be mediated by inhibition of JNK and NF-kappaB signal transduction.

To study the anti-fibrotic effects of danshensu, baicalin, astragalus and Panax notoginseng saponins (PNS) and their possible mechanisms.The four Chinese herb products mentioned above were given intraperitoneally to experimental rats with hepatic fibrosis. Colchicine was administered to a control group. Comparisons were made in four aspects: (1) Degrees of liver fibrosis; (2) Serum levels of hyaluronic acid (HA) and type IV collagen (CIV), using radioimmunoassay; (3) Densities of malondialdehyde (MDA), superoxide dismutase (SOD) and hydroxyproline (Hyp), using chromatometry, to detect the expression of tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), matrix metalloproteinase-1(MMP-1) and transforming growth factor beta 1 (TGF beta 1) in liver tissues, using immunohistochemical techniques; and (4) For hepatic stellate cells (HSCs): proliferation using MMT calorimetric assay, the cell cycles using flow cytometry, apoptosis using AO/EB fluorescence staining and type I and type III collagens using immunocytochemical stainings.(1) Compared with the model group, the serum levels of HA and CIV decreased significantly in all four drug-treated groups, especially in the danshensu-treated group. Astragalus and baicalin had better effects over PNS (P0.05 or 0.01). (2) In contrast to the model group, all four drugs dramatically reduced the amount of Hyp and MDA, increased SOD activity and reduced the degrees of liver fibrosis and the expressions of TIMP-1 and TGFbeta1 in liver tissues (P0.05 or 0.01). Danshensu had the best effect, astragalus and baicalin had similar effects which were stronger than PNS. (3) All four drugs inhibited HSCs proliferation, induced HSCs apoptosis and decreased type I, III collagen synthesis of HSC.The four drugs could minimize the hepatic fibrosis of rats in different degrees. Danshensu had the best effect, astragalus and baicalin had similar effects. The possible mechanisms of these effects might be related to inhibiting actions on activation and proliferation, promoting apoptosis and lowering the expression of type I and type III collagen of HSCs by down-regulating the expression of TGFbeta1; the decrease in the amount of MDA and the increase of SOD activity; and the reduction of extracellular matrix by down-regulation of TIMP-1/MMP-1.

To investigate the effect of Astragalus Injection solution on rat hepatic stellate cells (HSC) and hepatic fibrosis.HSCs of rats were incubated with various concentrations of Astragalus Injection solution (0 mg/ml, 25 mg/ml, 50 mg/ml, 100 mg/ml, 200 mg/ml, 400 mg/ml) for 24, 48 and 72 hours. Cell proliferation was detected with 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphennyltetrazolium bromide (MTT) colorimetric assay. Cell cycle was detected with flow cytometry. Cell apoptosis was detected with acridine orange/ethidium bromide (AO/EB) fluorescent staining and flow cytometry. In vivo, rats were randomly allocated into a normal control group, a model control group and an Astragalus Injection group. Astragalus Injection (800 mg.kg-1.d-1) was administered to rats of the Astragalus Injection group. Rats of the model control group received saline. Serum concentrations of hyaluronic acid (HA) and laminin (LN), hepatic tissue activity of superoxide dismutase (SOD), and hepatic tissue contents of malondialdehyde (MDA) were measured in these groups at 8 weeks. Hepatic tissue expression of LN was assessed by using immunohistochemistry. The pathological changes of hepatic tissues were examined by hematoxylin-eosin (HE) and van Gieson (VG) staining of their histological slides.In vitro, compared with the 0 mg/ml group, the proliferation of HSCs in other concentration groups was significantly inhibited by Astragalus Injection solution in a dose and time dependent manner, the cell proliferation cycle of HSCs was blocked in the G2-M phase, there was no apoptosis of HSCs in AO/EB fluorescent staining and flow cytometry. In vivo, compared with rats of the model control group, the rats of the Astragalus Injection solution treated group had remarkably decreased serum HA and LN levels (114.3+/-25.6) microg/L vs (85.6+/-37.3) microg/L and (78.8+/-11.7) microg/L vs (66.8+/-17.6) microg/L, P0.05, and liver MDA level (3.7+/-0.4) micromol/g protein vs (2.4+/-0.2) micromol/g protein, P0.01, but had increased activity of liver SOD (49.6+/-5.7) NU/mg protein vs (75.9+/-5.9) NU/mg protein, P0.01. Microscopic studies revealed that the livers of rats receiving Astragalus Injection solution showed decreases in fibrosis and in expression of LN.Astragalus Injection solution has an inhibitive effect on experimental hepatic fibrogenesis. The mechanisms of its effects might possibly be associated with its antioxidant activity, expression of decreasing LN and its inhibition of HSCs proliferation.

To investigate the effect and mechanism of Tanshensu on experimental fibrotic rats.Pigs serum was used to induce liver fibrosis in Wistar rats. The rats in Tanshensu-treated group were injected peritoneally with Tanshensu solution at the dose of 300 mg x kg(-1) x d(-1), and the rats in model-control group and normal-control group received the same volume of double distill water. At the end of the twelfth week, the hepatic stellate cells (HSCs) were isolated from the liver of one rat in model-control group using in-situ perfusion with pronase and collagenase, then density gradient centrifugation, and the other rats were killed to take the serum and liver samples. MTT colorimetric assay was used for detecting the proliferation of HSCs and flow cytometry was used for observing the cell cycles of HSCs under different concentrations of Tanshensu. The hyaluronic acid (HA) level in serum was detected and the morphological changes of liver tissue were observed.There was a decline of serum HA level in Tanshensu-treated group compared to that of the model-control group (231.4 ng/ml +/- 41.1 ng/ml vs. 398.7 ng/ml +/- 54.5 ng/ml, F =154.796, P0.05). Both HE and VG stain showed a decline of liver fibrosis degree in Tanshensu-treated group. And Tanshensu had an inhibition effect on the proliferation of HSCs at the concentrations of 50 mg/L, 100 mg/L, and 200 mg/L (1.60x10(-2) +/- 8.17x10(-4), 1.10x10(-2) +/- 1.41x10(-3), and 6.75x10(-3) +/- 3.30x10(-3) vs. 7.18x10(-2) +/- 1.71x10(-3), F =1154.221, P0.01).Tanshensu shows a therapeutic effect on liver fibrosis in rats induced by pig's serum through inhibiting the proliferation of hepatic stellate cells.

AIM: To observe expression of CD14 protein and CD14 gene in rat liver sinusoidal endothelial cells (LSECs) during endotoxemia, and the role of CD14 protein in the activation of lipopolysaccharide (LPS)-induced LSECs. METHODS: Wistar rat endotoxemia model was established first by injection of a dose of LPS (5 mg/kg, Escherichia coli O111:B4) via the tail vein, then sacrificed after 0 h, 3 h, 6 h, 12 h, and 24 h, respectively. LSECs were isolated from normal and LPS-injected rats by an in situ collagenase perfusion technique. The isolated LSECs were incubated with rabbit anti-rat CD14 polyclonal antibody, then stained with goat anti rabbit IgG conjugated fluorescein isothiocyanate (FITC) and flow cytometric analysis (FCM) was performed. The percentage and mean fluorescence intensity (MFI) of CD14-positive cells were taken as the indexes. LSECs were collected to measure the expression of CD14 mRNA by in situ hybridization analysis. The isolated LSECs from normal rats were incubated firstly with anti-CD14 antibody, then stimulated with different concentrations of LPS, and the supernatants of these cells were then collected for measuring the levels of tumor necrosis factor (TNF)-α and Interleukin (IL)-6 with ELISA. RESULTS: In rats with endotoxemia, LSECs displayed a strong MFI distinct from that of control rats. CD14 positive cells in rats with endotoxemia were 54.32%, 65.83%, 85.64%, and 45.65% at 3 h, 6 h, 12 h, and 24 h respectively, there was significant difference when compared to normal group of animals (4.45%) (P < 0.01). The expression of CD14 mRNA in isolated LSECs was stronger than that in control rats. In LPS group, the levels of TNF-α and IL-6 were 54 ± 6 ng·L-1, 85 ± 9 ng·L-1, 206 ± 22 ng·L-1, 350 ± 41 ng·L-1, 366 ± 42 ng. L-1 and 103 ± 11 ng·L-1, 187 ± 20 ng·L-1, 244 ± 26 ng·L-1, 290 ± 31 ng·L-1, and 299 ± 34 ng·L-1, respectively at different concentration points. In anti-CD14 group, the levels of TNF-α and IL-6 were 56 ± 5 ng·L-1, 67 ± 8 ng·L-1, 85 ± 10 ng·L-1, 113 ± 12 ng·L-1, 199 ± 22 ng·L-1 and 104 ± 12 ng·L-1, 125 ± 12 ng·L-1, 165 ± 19 ng·L-1, 185 ± 21 ng·L-1, and 222 ± 23 ng·L-1, respectively at different concentration points. There was significant difference between the two groups (P < 0.01). CONCLUSION: LSECs can synthesize CD14 protein and express CD14 gene during endotoxemia. CD14 protein plays an important role in the activation of LPS-induced LSECs. This finding has important implications for the understanding of the mechanisms by which LPS may injure liver sinusoidal endothelial cells during sepsis.

Water salinization in semi-arid and arid regions is threatening freshwater or oligosaline ecosystems. Anthropogenic processes enrich nutrients of aquatic systems causing significant environmental effects. Bosten Lake in Xinjiang Province, China is an interesting ecosystem featuring a salinity gradient from fresh to subsaline, as well as a nutrition gradient from oligotrophic to mesotrophic. In the present study, we focused on the effects of salinity and nutrients in Bosten Lake by investigating the zooplankton assemblages and environmental factors from different sampling times. A total of 74 zooplankton taxa were found, consisting of 34 rotifers, 26 protists, 10 cladocerans, and 4 copepods. Although no significant differences were found among different sampling sites, zooplankton species richness, diversity, and evenness showed significant differences between sampling times, with August samples showing highest values along with water temperature and total nitrogen. Principal component analysis and representational difference analysis results showed that zooplankton abundance is correlated with water temperature and nitrogen, but showed no significant relationship with water mineralization or conductivity. The subsaline-tolerating zooplankton species in Bosten Lake made their community insensitive to salinity. The decrease of total nitrogen concentration in Bosten Lake probably implied a primary productivity increase, which subsequently caused the zooplankton diversity to increase in August.

AIM: To observe the synthesis of endotoxin receptor CD14 protein and its mRNA expression in Kupffer cells (KCs), and evaluate the role of CD14 in the pathogenesis of liver injury in rats with alcohol-induced liver disease (ALD). METHODS: Twenty-eight Wistar rats were divided into two groups: ethanol-fed group and control group. Ethanol-fed group was fed ethanol (dose of 5-12g·kg·d-1) and control group received dextrose instead of ethanol. Two groups were sacrificed at 4 wk and 8 wk, respectively. KCs were isolated and the synthesis of CD14 protein and its mRNA expression in KCs were determined by flow cytometric analysis (FCM) or the reverse transcription polymerase chain reaction (RT-PCR) analysis. The levels of plasma endotoxin and alanine transaminase (ALT) were measured by Limulus Amebocyte Lysate assay and standard enzymatic procedures respectively, and the levels of plasma tumor necosis factor (TNF)-α and interleukin (IL)-6 were both determined by ELISA. The liver pathology change was observed under light and electric microscopy. RESULTS: In ethanol-fed group, the percentages of FITC-CD14 positive cells were 76.23% and 89.42% at 4 wk and 8 wk, respectively. Compared with control group (4.45% and 5.38%), the difference was significant (P < 0.05). The expressions of CD14 mRNA were 7.56 ± 1.02 and 8.74 ± 1.37 at 4 wk and 8 wk, respectively, which were significantly higher compared with the control group (1.77 ± 0.21 and 1.98 ± 0.23) (P < 0.05). Plasma endotoxin levels at 4 wk and 8 wk increased significantly in ethanol-fed group (129 ± 21 ng·L-1 and 187 ± 35 ng·L-1) than those in control rats (48 ± 9 ng·L-1 and 53 ± 11 ng·L-1)(P < 0.05). Mean values of plasma ALT levels increased dramatically in ethanol-fed rats (112 ± 15 IU/L and 147 ± 22 IU/L) than those in the control animals (31 ± 12 IU/L and 33 ± 9 IU/L) (P < 0.05). In ethanol-fed rats, the levels of TNF-α were 326 ± 42 ng·L-1 and 402 ± 51 ng·l-1 at 4 wk and 8 wk, respectively which were significantly higher than those in control group (86 ± 12 ng·L-1 and 97 ± 13 ng·L-1) (P < 0.05). The levels of IL-6 were 387 ± 46 ng·L- 1 and 413 ± 51 ng·L-1, which were also higher than control group (78 ± 11 ng·L-1 and 73 ± 10 ng·L-1) (P < 0.05). In liver section from ethanol-fed rats, there were marked pathological changes including steatosis, cell infiltration and necrosis. No marked pathological changes were seen in control group. CONCLUSION: Ethanol administration led to a significant synthesis of endotoxin receptor CD14 protein and its gene expression in KCs, which maybe result in the pathological changes of liver tissue and hepatic functional damages.