Objective: To study bacterial and lactobacilli communities in stomach of neonatal piglets from 7 to 35(two weeks after weaning) days of age.Methods: The semi-quantitative Dilution-PCR,PCR-DGGE and sequencing were used to analyze the shifts of stomach total bacterial and lactobacilli community.Results: Stomach pH value was around 3.0 before weaning and increased apparently after weaning.Dilution-PCR showed that bacterial concentration in stomach of piglet was very low before weaning and increased significantly after weaning.The DGGE profiles from total bacteria in stomach showed that the bands of samples before weaning were very simple and the predominant band belonged to high GC bacterium.After weaning,some new predominant bands appeared and the previous high GC bacteria disappeared simultaneously.Sequence analysis indicated that these new bands were Escherichia fergusonii,Escherichia coli,Streptococcus gordonii and Streptococcus suis,all of which were potential pathogens for piglets.The similarity of DGGE profiles from stomach lactobacilli showed weaning stress had no obvious effect on lactobacilli community.Conclusion: Weaning stress could cause stomach pH increase apparently and induce potential pathogens proliferation in stomach of piglets.

With the improvement of living conditions,people pay more attention to food sanitation,following which the incidence rate of food allergy is higher and higher.Food allergy threatens public health seriously.It is caused mainly by dominance of T helper type 2-based immune responses,which breaks the dynamic balance of Th1/Th2 immune responses and resultes in over secreting of IL-4,IL-5 and IL-13,production of specific antibody IgE,degranulation of mast cells,alteration of intestinal microflora and so on.At present,a lot of investigations have been reported,of which the probiotics therapy attracted more attention.As the indigenous flora,probiotics could not only prevent and treat food allergy but could also modulate immune responses and recover the dynamic balance of it via secreation of cytokines such as IFN-?,IL-10,TGF-?.

Dietary essential amino acids are extracted in the first-pass metabolism by the small intestine.Besides incorporation into mucosal protein,they are also the fuels for the utilization by small intestinal mucosal cells and the precursors for intestinal synthesis of new amino acids and enormous biological important molecules such as glutathione,polyamines,etc.Thus,their gut metabolism is not only crucial for the maintenance of the integrity and function of intestinal mucosa,but also important for the regulation of whole body metabolism.

Objective To investigate the effect of dexamethasone combined with oridonin on proliferation and apoptosis in multiple myeloma cells U266 and the related molecular mechanism. Methods Exponential phase of growth U266 cells were treated with different concentrations of oridonin combined with dexamethasone or alone. U266 cells treated by DMSO were taken as control group. The proliferation inhibitory ratios were measured by CCK-8 assay followed by 24 h, 48 h and 72 h. Apoptosis induction was assessed by using Annexin V-FITC kit. Real time PCR was used to examine the mRNA changes of Notch1, NF-κB/p65 and bcl-2. Western blot assay was applied to detect the protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2. Results Compared with that in control group, proliferation in all the experimental groups was inhibited (P<0.05), and the apoptosis was promoted (P<0.05); especially the combination of dexamethasone and oridonin had a synergistic effect on the proliferation and apoptosis of U266 cells (P<0.05). The results of PCR and Western blot showed that after treatment of U266 cells with dexamethasone, the mRNA as well as their protein levels of NF-κB/p65 and bcl-2 were decreased compared with those in the control group (P<0.05). Moreover, the mRNA and protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2 was obviously down-regulated in oridonin group and the combination group (P<0.05). Conclusion Combination of dexamethasone and oridonin can significantly increase the anti-tumor effect by inhibiting proliferation and inducing apoptosis of U266 cells, which may be related to the inhibition of the Notch1 pathway.

BACKGROUND:It is reported that the cellsurface antigen may be damaged by the trypsin enzyme in some extent and the cellactivity may also be influenced, as a result of which, the subsequent separation and fol ow-up-test wil be affected. Accutase enzymes possess the activities of protease and col agen enzyme and need no special termination or cleaning at the end of digestion. Another special function of accutase enzyme is to protect cellsurface antigen and thus it has been widely applied in the stem celldigestion and culture.
OBJECTIVE:To compare the digestive effect of accutase enzymes and trypsin enzyme in the separation and original culture of spermatogonial stem cells.
METHODS:The testes of 5-7 days male Kunming mice (n=45) were col ected and primarily digested with col agenas. The suspension of digested tissue was divided into three parts with the same volume named accutases enzyme group, trypsin enzyme group and mixed enzyme group (trypsin enzyme and hyaluronidase). For the comparison of testis digestive status and the time required for the formation of single cells, the micrographs were taken at 1, 3 and 5 minutes respectively after enzyme digestion. The total number and the mortality of the single cells were estimated and compared. The leydig cells and sertoli cells were removed by differential adherent method and the remained spermatogenic cells were then treated with CD90.2 immune magnetic beads. The selected spermatogonial stem cells were subsequently labeled by glial cellline-derived neurotrophic factor receptor alpha-1 and sorted with flow cytometry. The number of spermatogonial stem cells positive for glial cellline-derived neurotrophic factor receptor alpha-1 in CD90.2+and CD90.2-cells was compared within different groups.
RESULTS AND CONCLUSION:The digestive capacities of different enzyme were different. Accutases enzyme obtained the single cellsuspension more quickly than trypsin enzyme and mixed enzyme, and had the least cellmasses and broken cellmembrane than the other two groups. After the differential attached treatment, the highest total number of CD90+spermatogonial stem cells and lowest cellmortality could be found in the accutases group, when compared with the other groups. The results of cellsorting by flow cytometry showed a higher rate (72.24%) of GFRα1+/CD90+cells in the accutases group than the trypsin group (51.16%) and mixed enzyme group (71.27%). The GFRα1+/CD90-number in the accutases group was lower (15.03%) than that of the trypsin group (18.8%) and mixed enzyme group (24.23%), respectively. The results indicate a better effect of accutases enzyme on the primary separation of spermatogonial stem cells than that of trypsin or mixed enzyme.

Objective A rat model of chronic post-thoracotomy pain is used to study whether acute pain and pre-operative diffused noxious inhibitory controls(DNIC) can predict chronic pain and how DNIC changes when pain maintains.Methods Rats were randomly divided into three groups:naive group,sham group and model group.DNIC was constantly assessed in individual rats,along with each animal's mechanical hyperalgesia and cold allodynia after thoracotomy.Results In model group,the incidence of chronic post-thoracotomy pain was 55%(11 of 20),which was named CPTP group,and the other 9 rats without chronic pain was defined as non-CPTP group.The pre-operative DNIC was significantly weaker in CPTP group with lower mechanical threshold on 6 days after surgery and higher cold sensitivity on 6 days after surgery comparing with non-CPTP group.In the acute pain phase (day 3),DNIC was decreased in both CPTP group and non-CPTP group as compared with pre-operative period.Besides,DNIC was recovered in non-CPTP group while kept impaired in CPTP group on 21 days after surgery.Conclusions Pre-operatively assessed DNIC efficiency and acute post-operative pain intensity were two independent predictors for CPTP.DNIC was decreased both in acute pain and chronic state,while returned to normal when pain sense was normal.

OBJECTIVE:To study the effects of Xinghuang tablets on tumor growth and immunologic function in S 180 sarcoma-bearing mice.METHODS:The model of mice with S 180 sarcoma were given Xinhuang tablets intragastrically to ob?serve the effects on the ratio of tumor to body,the inhibition rate of tumor,the indices of the spleen and thymus,and the rate of micronuclei in PCE in bone marrow of the mice.RESULTS:The body weight of the mice with S 180 sarcoma was decreased;the growth of the tumor was inhibited;the weight of spleen was increased;the number of leucocyte of peripheral blood was raised(P

Inferior vena caval (IVC) tumor thrombus with cardiac extension is a very rare phenomenon,which proliferates fast and could be very challenging to the surgery.This paper was designed to investigate the clinical value of transesophageal echocardiography (TEE) for the surgical resection of IVC tumor thrombus extending into right cardiac cavities.Six cases from our medical institute,preoperatively diagnosed as IVC tumor thrombus with cardiac extension and scheduled for the surgical resection,were retrospectively analyzed.In addition to real-time and dynamic monitoring,comprehensive TEE exams were performed for all the patients respectively after anesthesia induction,namely before tumor resection and after tumor resection.Cardiac extension was defined by the preoperative finding of cardiac mass originated from IVC tumor thrombus by transthoracic echocardiography,computerized tomography or CT angiography.In all the cases,intraoperative TEE provided an accurate and excellent view of the IVC tumor thrombus.For case three,the IVC tumor thrombus was found at the IVC entrance to right atrium without further cardiac extension; for case five,the IVC tumor thrombus proliferated into right atrium but the extended cardiac mass was very slim and flexible and the tricuspid valve was untouched; for case four,the IVC tumor thrombus extended into right atrium and even cross the tricuspid valve but the extended cardiac mass was also very slim and flexible.Based on the TEE-provided information,the originally scheduled surgical decision was modified and the surgical resection was performed without cardiopulmonary bypass (CPB).For the other three cases,the intraoperative TEE showed similar results to preoperative findings.The huge IVC tumor thrombus extended into the right heart,presented almost no flexibility and dramatically compromised the intracardiac blood flow.For the three cases,CPB support was indispensable for the tumor resection.The full TEE exam after tumor resection in all the six patients displayed clear surgical resection without tumor residuals,but in those three patients suffered with severely compromised cardiac extension,severe tricuspid regurgitation was noticed.All the six patients were closely monitored until discharged,and no TEE-related complications were observed.This paper reports about TEE' s utilization in a series of consecutive patients undergoing surgical resection of IVC tumor thrombus with cardiac extension.In addition to its safety and effectiveness,TEE can provide valuable information for surgical decision making,surgical intervention assessment and anesthesiamanagement strategies.

Objective To investigate the application of contrast-enhanced ultrasonograhy CEUS in the early assessment of antiangiogenic treatment in metastatic colorectal cancer Methods Among 41 colorectal cancer patients with liver metastases enrolled in this study 25 patients received treatment of antiangiogenic treatment bevacizumab combined with chemotherapy BV group and 16 patients received chemotherapy only non-BV group To assess the treatment responses in the liver lesions between and within these two groups CEUS scans were performed before the treatment T0 or??baseline on the day 2 T1 and on the day 7 T2 after the treatment The percentage differences were evaluated according to the CEUS quantitative parameters such as rise time RT peak intensity PI mean transit time MTT area under the curve AUC time from peak to one half TPH wash in slope WIS and time to peak TTP Results On T1 both the RT and MTT increased within the BV group and decrease within the non-BV group when compared with their own baselines the differences were statistically significant P <0 05 On T2 the MTT increased within the BV group and decreased within the non-BV group when compared with their baselines the differences again were statistically significant P <0 05 All other parameters measured on T1 and T2 did not show a significant difference when compared with baselines Within the BV group the RT decreased and WIS and TPH increased for the responding subjects when compared with the non-responding subjects the differences were statistically significant P < 0 05 Within the non-BV group however no statistically significant differences were found for all parameters between the responding subjects and non-responding subjects P >0 05 Conclusions CEUS can be used to assess the treatment response in liver metastatic patients with colorectal cancer by monitoring the change of the tumor perfusion before and after the treatment.

This study aimed to elucidate the effect of tryptophan (Trp) on gut hormone secretion as well as the roles of the calcium-sensing receptor (CaSR) and its downstream signaling pathway in gut hormone secretion by assessing swine duodenal perfusion in vitro. Swine duodenum was perfused with Krebs-Henseleit buffer as a basal solution. Various concentrations (0, 10, and 20 mM) of Trp were applied to investigate its effect on gut hormone secretion. A CaSR antagonist was used to detect the involvement of CaSR and its signal molecules. The 20 mM Trp concentration promoted the secretion of cholecystokinin (CCK) and glucose-dependent insulinotropic peptide (GIP), elevated the mRNA level of CaSR, and upregulated the protein levels of CaSR, protein kinase C (PKC), and inositol trisphosphate receptor (IP3R). However, NPS 2143, an inhibitor of CaSR, attenuated the CCK and GIP release, reduced the mRNA level of CaSR, and decreased the protein levels of CaSR, PKC, and IP3R with 20 mM Trp perfusion. The results indicate that CCK and GIP secretion can be induced by Trp in swine duodenum in vitro, and the effect is mediated by CaSR and its downstream signal molecules PKC and IP3R.
Cholecystokinin ; Duodenum ; Gastric Inhibitory Polypeptide ; In Vitro Techniques ; Inositol ; Perfusion ; Protein Kinase C ; Receptors, Calcium-Sensing ; RNA, Messenger ; Swine ; Tryptophan