ObjectiveTo explore the mechanism of Shangerhuang Wan （SEHW） and its subdivisions in alleviating loperamide-induced functional constipation （FC） in rats by regulating bile acid metabolism. MethodsSixty rats were randomly assigned into six groups （n=10）： normal control， model， positive control （domperidone， 8 mg·kg^-1）， Cimicifugae Rhizoma-Bupleuri Radix （SC， 1.33 g·kg^-1）， Scutellariae Radix-Coptidis Rhizoma （QL， 1.20 g·kg^-1）， and SEHW （3.33 g·kg^-1）. The remaining groups except the normal control group were subjected to subcutaneous injection with loperamide at 3 mg·kg^-1 twice daily （total dose of 6 mg·kg^-1·d^-1） for 7 days to induce a model of FC. Drug administration was initiated on day 3 of modeling， which continued throughout the modeling period. The small intestinal propulsion rate of each group was measured via the ink propulsion method. LC-MS/MS was employed to measure the fecal bile acid content in each group， and the serum bile acid level was measured by a microplate. The 5-hydroxytryptamine （5-HT） and cyclic adenosine monophosphate （cAMP） levels in the colon tissue of each group were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was employed to determine the protein levels of apical sodium-dependent bile acid transporter （ASBT） in the ileum terminus and Takeda G protein-coupled receptor 5 （TGR5） in the colon. ResultsCompared with the normal control group， the model group exhibited decreases in the small intestinal propulsion rate and total fecal bile acid content （P<0.01）， an elevation in the serum total bile acid level， lowered 5-HT and cAMP levels （P<0.01）， up-regulation in the protein level of ASBT （P<0.01）， and down-regulation in the protein level of TGR5 （P<0.05）. Compared with the model group， the positive control， SEHW， and QL groups showed increases in the small intestinal propulsion rate （P<0.05， P<0.01） and total fecal bile acid content （P<0.01） and a decline in the serum bile acid level. Compared with the SEHW group， the SC group had decreased total fecal bile acid content （P<0.01） and an elevated serum bile acid level， while the QL group showed increased total fecal bile acid content （P<0.01） and a lowered serum bile acid level. Compared with the model group， the positive control， SEHW， and QL groups demonstrated down-regulation in the protein level of ASBT （P<0.05， P<0.01） and up-regulation in the protein level of TGR5 （P<0.05， P<0.01）. The SC group showed no significant change in the protein level of ASBT and up-regulation in the protein level of TGR5 compared with the model group （P<0.05）. Additionally， compared with the model group， the positive control， SEHW， and QL groups showed elevated 5-HT and cAMP levels （P<0.05， P<0.01）. The SC group showed no significant difference in the 5-HT level but a rise in the cAMP level （P<0.01）. ConclusionSEHW， through the compatibility of QL and SC， demonstrates a dual regulatory effect on bile acid metabolism， embodying the principle of descending turbidity and ascending lucidity， which highlights the compatibility and scientific rationale of this formula. SEHW inhibits ASBT expression in the ileum to reduce ileal bile acid reabsorption， increase colonic bile acid content， and bind to colonic TGR5 to release 5-HT， thereby enhancing intestinal motility and promoting intestinal contraction.

[ ABSTRACT ] AIM: To observe the effect of rapamycin on the apoptosis of mouse astrocytes in vitro.ME-THODS:The astrocytes from C57BL/6J newborn mouse pups were isolated and primarily cultured.The effect of rapamycin on the viability of astrocytes was assessed by MTT assay.The mean fluorescence intensity of SYTOX?Green stain in the astrocytes was detected by fluorescence microplate reader in order to analyze the effects of rapamycin on the cell death in-duced by H2 O2 , ionomycin and/or deferorxamin.DiOC6 (3) staining was used to analyze the mitochondrial membrane po-tential of the astrocytes induced by H2 O2 .Flow cytometry analysis was used to determine the production of ROS in the as-trocytes and mitochondria by staining with H2 DCFDA and MitoSOXTM Red reagent, respectively.RESULTS: Rapamycin at concentration of 0.5 μmol/L protected the astrocytes against cell death induced by H2 O2 or deferoxamine plus ionomy-cin.Rapamycin protected the mitochondrial membrane potential of astrocytes from the injury of H2 O2 .It also reduced the production of ROS in the astrocytes and decreased the level of ROS in the mitochondria.CONCLUSION:Rapamycin re-duces the ROS overload in the mitochondria, keeps mitochondrial membrane potential safety and protects the astrocytes a-gainst apoptosis in vitro.

BACKGROUND:Because macrophages play an important role in the body’s inflammatory response, the detection of the impact of biological materials on the behavior of macrophages can assess the immunogenicity of materials. OBJECTIVE: To analyze the activation effect of decelularized extracelular matrix materials on macrophages. METHODS: The peritoneal macrophages of BALB/c mouse were obtained and cultured by dividing into five groups. Control group was simple cel culture group, experimental group 1 was acelular matrix membrane material directly contacting with macrophage for culture, experimental group 2 was fresh pericardial material directly contacting with the macrophage for culture, experimental group 3 was acelular matrix membrane material indirectly contacting with macrophages for culture, experimental group 4 was fresh pericardium material indirectly contacting with macrophages for culture. After 24 hours of culture, the secretion of nitric oxide and cytokines in cel culture supernatant was determined. After 48 hours of culture, the absorbance value was determined by MTT method and the toxicity grading was determined. RESULTS AND CONCLUSION: The toxicity grading in experimental groups 1-4 was respectively grades 2, 4, 0, 2. The nitric oxide level in experimental groups 1 and 2 was higher than that in the control group (P < 0.05), and the nitric oxide level in the experimental group 2 was higher than that in the experimental group 1 (P < 0.05). There were no significant differences in interleukin-2, interleukin-4,interferon γ, interleukin-17A and interleukin-10 levels between these five groups. The interleukin-6 level in the experimental group 2 was higher than that in the control group (P < 0.05); The expression levels of tumor necrosis factors in experimental group 1, 2 and 4 were higher than those in the control group (P < 0.05), and experimental group 2 higher than the experimental group 1 (P < 0.05), experimental group 1 higher than the experimental group 4 (P < 0.05). These results show that acelular matrix material can activate macrophages in direct contact.

Before the technique of advanced high-throughput sequencing comes up , less is known about the human gut microbiota .It has been understood that trillions of microbes , in which 99% are bacteria , inhabit the human gut, forming a complicated ecological community .The gut microbiota has a great impact on human physiology and suscepti -bility to disease through its integrative metabolic activities and interactions with the host .In physiology , gut microbiota con-tributes to the host acquisition of nutrition and energy from diets , promoting development and maturation of gastrointestinal tract and immune system , and protecting host from invasion of enteropathogens .In pathology , dysbiosis underlying altered gut microbiota is associated with the susceptibilities to various diseases , including inflammatory bowel disease , type 1 dia-betes, asthma, obesity, metabolic syndrome , autism and cancer .Understanding of the factors that underlie alterations in the composition and function of gut microbiota will be helpful in the development of drugs and the design of therapies that target it.This goal is formidable .It is because that the compositions of gut microbiota are immensely diverse , varying be-tween individuals in a population and fluctuating over time in an individual , especially during early development and disea-ses.Viewing the gut microbiota with an ecological perspective will provide new insights into how to improve our health by targeting this microbial community in clinical treatments .

Objective To research the effect of the identity recognition system based on the Itouch in the patients care .Methods The study group used the identification system of Itouch in all aspects of nursing intervention .Patients without this system were selected as the control group .The rate of operation error , time spent on writing and patient satisfaction rate were compared between groups .Results The error rates of intravenous infusion , oral medicine distribution , specimen collection intravenous injection and skin test were 0.58%,1.42%,0.45%,0.55%,0.62%in the study group, and 0.72%,2.23%,0.79%,0.64%,2.93%in the control group, there were significant differences between two groups (χ2 =5.21,38.37,6.39,7.26,4.85, respectively;P<0.05).The number of patients who were very satisfied , satisfied and dissatisfied was 685,506 and 55 in the study group and 566 , 566 and 146 in the control group , there were significant differences between two groups (χ2 =55.48,P<0.01).The time nurses spent on writing , signature, changing orders and checking were (24 ±12)min,(16 ±3)min,(14 ±5)min in the study group, and (206 ±42)min,(72 ±6)min,(69 ± 17)min in the control group.There were significant differences between two groups (t=22.82,45.72,17.00, respectively;P <0.01).Conclusions The identity recognition system based on the Itouch was helpful to improve the accuracy rate , work efficiency and the satisfaction rate of patients and to reduce the nursing adverse events.

Objective To detect systematic oxidative stress in preeclampsia.Methods (1)Morphological features of placenta hypoxia were observed by histological method ; (2) Level of granulocyte intracellular reactive oxygen species was monitored by dyeing full blood with 2' ,7'-dichlorodihydrofluorescein diacetate (H2DCFDA) ; (3) Level of H_2O_2 in sera was detected by special kits.Results Compared to normal pregnancy,placentas from preeclampsia showed distinct features of hypoxic stress injury,such as more syncytial knots formation,fibrosis emerged,vein in-jury and loss its normal configuration; Fluorescence values of ROS probe in neutrophils from different women were 45.61±12.20(n =49),51.02 ± 13.60(n =56,P <0.01)and 85.10 ± 16.30(n =47,P <0.01); Concentra-tions of H_2O_2were (24.57±5.17)μmol/L(n =49),(26.61±3.25)μmol/L(n =56,P 0.01) and (39.84±9.67)μmol/L(n=47,P<0.01) respectively.Conclusion With the help of histological method,flow cytometry and special kits,systematic oxidative stress can be detected through checking placentic tissues,netrophils and sera of preeclampsia.

AIM: To investigate the anti-HIV-1 activity of five anthraquinone derivatives (emodin,rhein,chrysophanol,physcion and aloe-emodin) in vitro.METHODS: Viral replication was estimated by observation of cytopathogenesis and measurement of HIV-1 p24 antigen production in HIV-1ⅢB acutely infected C8166 cells. The anti-HIV-1 activity was evaluated by the 50% effective concentrations (EC50) and selective indexes (SI) of these derivatives.RESULTS: These anthraquinone derivatives inhibited HIV-1ⅢB replication on syncytia formation induced by HIV-1ⅢB infection with EC50 mean values of (11.44±0.93)μmol/L (emodin),(51.28±2.86)μmol/L (rhein),(90.58±2.30)μmol/L (chrysophanol),(8.59±0.38)μmol/L (physcion) and (0.89±0.08)μmol/L (aloe-emodin),respectively. The p24 antigen production with EC50 mean values were (11.61±0.56)μmol/L (emodin),(12.35±4.73)μmol/L (rhein),(39.63±2.87)μmol/L (chrysophanol),>250 μmol/L (physcion) and (2.75±0.20)μmol/L (aloe-emodin) respectively. CONCLUSION: These structurally-related chemicals show different anti-HIV-1 activity in vitro. Among them,aloe-emodin is the most potent inhibitor to HIV-1 replication.

AIM: To investigate the effect of dihydroartemisinin (DHA) on the proliferation of murine T lymphocytes stimulated by Con A in vitro and its related immunosuppressive mechanism. METHODS: Murine T lymphocytes were stimulated by Con A and treated with different concentrations of DHA. Cell proliferation was measured by carboxyl fluoresce in diacetate succinmidyl ester (CFDA-SE) staining. The expression of CD69, CD25 and CD71,which was the marker of early, middle, later activation of CD3~+ T lymphocytes, was measured by flow cytometry (FCM) combined with two-color immunofluorescent staining of cell surface antigen. Fluorescence calcium indicator fluo-4/AM was used to measure the change of the intracellular calcium concentration ([Ca~(2+)]_i) of murine T lymphocytes. The distribution of the cell cycle was analyzed by PI staining. The expression of CD69, the early activation antigen on CD4~+CD25~(high) Treg was also measured by FCM combined with three-color immunofluorescent staining. RESULTS: The result of CFDA-SE staining showed that DHA efficiently inhibited the Con A-induced proliferation of T-lymphocytes in a time-and dose-dependent manners. DHA showed modestly increased proportions of CD69 and CD25 on Con A-stimulated CD3~+T cells, but inhibited the expression of CD25 in a dose dependent manner. DHA with Con A, but not DHA alone, caused an increase in intracellular calcium concentration of T cells. The results of FCM analysis with PI staining showed that DHA imposed a total cell cycle arrest in G_0/G_1 and prevented cells entering S phase and G_2/M phase. Furthermore, DHA reduced the expression of CD69 on CD4~+CD25~(high) Treg. CONCLUSION: DHA, which exhibits immunosuppressive effect on the proliferation of murine T-lymphocytes, is promising to be developed as an immunosuppressive reagent.

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.

BACKGROUND: The connection between Natural killer(NK)-cells and allogeneic bone marrow transplantation(allo-BMT)has aroused increasing attention.OBJECTIVE: To explore the effect of NK cells on graft rejection,hematopoietic and immune reconstitution in mouse undergoing allo-BMT.METHODS: Lethally and nonlethally irradiated BALB/c(H-2d)mice were transplanted with C57BL/6(H-2b)bone marrow plus donor peripheral T cells and/or NK cells.RESULTS AND CONCLUSION: Compared with lethally irradiated and allo-BMT group without infusion of NK cells,the survival rate in lethally irradiated and allo-BMT group with infusion of NK cells significantly enhanced; leukocytes count,expression level of CD19+and CD34+cell count recovered rapidly; expression level of H-2b*cell obviously increased.Expression level of CD34"cell in the group with infusion of NK cells was obviously lower than that of the group without infusion of NK cells at 28 days after transplantation,but there was no significant difference between the 2 groups at 60 days(P > 0.05).In nonlethally irradiated and allo-BMT group without NK cell infusion,expression level of H-2b*cell significantly decreased at 30 days after transplantation,and reduced to before transplantation level at 60 days; while expression of H-2b+cell yet could be detected with more than 80% at 60 days after transplantation in group infused with high and low concentration of NK cells.In alIo-BMT mice,alloreactive NK cell inhibits graft rejection,enhances engraftment,promotes the reconstitution of hematopoiesis and immunity,and increases survival rates.