Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

A pilot-scale research on purification of odorous gas emitted from wastewater treatment plant using a biofilter was conducted. The aim of this study is to check on the performance of biofilter running in various conditions and the effect of pH fluctuations on the performance of biofilter. The relation between distribution of microorganism and removal of odorous gases were also discussed here. The experimental results show that the predominant odor-causing gas can be efficiently eliminated by a biofilter inoculated with deodoring microorganism which were isolated previously. Moreover the biofilter had been proved having good tolerance to shocking loads of pollutant and can operate well in the condition of low pH.

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

In order to explore the dormancy physiological and biochemical mechanism of Paris seeds, the seed embryo growth courses, and the dynamic change of 5 enzymes, include SOD, POD, CAT, MDH, G-6-PDH were measured during variable temperature stratification. The results indicated that Paris seeds embryo grew quickly after 40 d in warm-stratification (18 ± 1) °C, at the meantime the metabolic activity was significantly strengthened. These facts showed that Paris seeds turned into physiological after-ripening process. After 60-80 d, the morphological embryo after-ripping process basically completed, and the following cold-stratification (4 ± 1) °C furthered Paris seed to finish physiological after-ripening. After 40 d, the activity of MDH decreased while G-6-PDH increased significantly. This showed that the main respiratory pathway of seed changed from TCA to PPP, which benifited breaking seed dormancy. In the whole period of stratification process, the activity variation of SOD and CAT was insignificantly and the activity of POD was enhanced significantly after shifting the seed in cold stratification process. This showed that SOD, CAT had no direct effects on breaking Paris seed dormancy but keeping the seed vigor, while the POD might involve in the process of Paris seed dormancy breaking.
Germination ; Liliaceae ; chemistry ; embryology ; enzymology ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; Temperature

Objective To observe the dynamic changes in mesenteric lymph microcirculation and its kinetics in rats suffering from severe heatstroke (SHS), and to explore the role of mesenteric lymph in the pathogenesis of SHS. Methods SHS rat models were reproduced in an incubator with high temperature and high humidity. The vital signs and the time of onset of SHS in rats were recorded continuously during the process of heat stress. Parameters of mesenteric lymph microcirculation including Index-I, Index-II, L.D-Index, and intra-lymphatic pressure before heat exposure, 60min after heat exposure, and onset of SHS were collected and analyzed. Mesenteric lymph was collected at 30-min interval, and its volume of production was measured dynamically. Results Rat SHS model was reproduced successfully. After exposure to the environment with high temperature and high humidity, the core temperatures of the rats raised to 42°C at the time point of 60min, and HS onset occurred at about 77min. Mesenteric lymph-vessel contraction indices including Index-I, Index-II, L.D-Index, lymph-vessel pressure and mesenteric lymph flow decreased significantly at the time point of 60min (P<0.05). However, all the above parameters increased at the time point of SHS onset (P<0.05), but had not yet reached the normal levels before hyperthermia and high humidity exposure (P<0.05). Conclusion Changes of mesenteric lymph microcirculation in rats with SHS shows a dynamical regularity, which may take part in the pathogenesis of SHS.

Objective To explore the effect of mesenteric lymph from rats suffering from heat stroke (SHS) on the inflammatory activity of vascular endothelial cells. Methods Rats were placed in a prewarmed incubator to reproduce heat stroke. Human umbilical vein endothelial cells (HUVECs ECV-340) were incubated in 5% mesenteric lymph collected pre-, during-, and post-HS for 3 and 6 h respectively in vitro. The levels of high mobility group protein B1 (HMGB1), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 of ECV-340 cells were determined. The expression of intercellular adhesion molecule 1 (ICAM-1) mRNA and NF-κB activity of ECV-340 were also assayed. Results The levels of HMGB1, TNF-α, IL-1β and IL-6 induced by SHS mesenteric lymph of rats significantly increased after heat treatment (P<0.05), and this effect was enhanced with prolongation of exposure time (P<0.05). The expression of ICAM-1 mRNA and NF-κB activity were significantly elevated after being activated by SHS mesenteric lymph (P<0.05). Conclusion HS mesenteric lymph probably activates the inflammatory responses of vascular endothelium, which is closely associated with the pathogenesis of SHS.

Studies concerning the association between arsenic exposure and hepatitis B virus (HBV) infection have been lacking.The present study aimed to examine the association between total urinary arsenic (TUA) and infection of HBV.A total of 5186 participants from National Health and Nutrition Examination Survey (NHANES) 2003-2014 were included in the analysis.We used logistic regression to evaluate the association.We defined two measures of TUA.TUA1 was the sum of arsenous acid,arsenicacid,monomethylarsonic acid and dimethylarsenic acid.TUA2 was defined as TUA minus arsenobetaine and arsenocholine.The results showed that the weighted overall prevalence of HBV infection was 6.08％.For NHANES 2003-2014,the medians (interquartile range) of TUA1 and TUA2 were 5.60 μg/L (3.97-8.09 μg/L) and 4.91 μg/L (2.36-9.11 μg/L),respectively.Comparing the highest quartile to the lowest quartile after multivariable adjustment showed that the odds ratios (ORs) and 95％ confidence intervals (CIs) for TUA1 and TUA2 were 2.44 (1.40-4.27) and 2.84 (1.60-5.05),respectively.In conclusion,elevated urinary arsenic was associated with the risk of HBV infection.Further studies,especially prospective studies,are needed to confirm the causal relationship between arsenic exposure and HBV infection.

Adult ; Castleman Disease ; complications ; diagnostic imaging ; pathology ; surgery ; Humans ; Lung Neoplasms ; etiology ; pathology ; Male ; Radiography

AIMTo study the effects of yi-zhi II (a compond of Chinese Traditional Medicine) on the alteration of synaptic structure in hippocampal CA3 and maintenance of memoy.

METHODSBy using the method of oral administration of yi-zhi II, the step-through test and electron microscopy, the latency of step-through and synaptic structure in hippocamal CA3 were tested.

RESULTS(1) The mice which had been given yi-zhi II prolong significantly the latency of step through (P < 0.05 or P < 0.01) on the 1st, 6th and 12th day after learning. (2) On the 6th and 12th day after learning, the length of synaptic active zone were markly improved in yi-zhi II and control, but that of yi-zhi II was better than that of control. (On the 6th day after learning, the number of perforated synapses and axo-dendrite synapses were significantly improved by the yi-zhi II (P < 0.05).

CONCLUSIONThe yi-zhi II could improve the learning and memory in mice. It migth improve the memory by increasing the length of synaptic active zone and the number of perforated synapses and axo-dendrite synapses in hippocampal CA3.

Animals ; Avoidance Learning ; drug effects ; CA3 Region, Hippocampal ; drug effects ; physiology ; Chromosome Pairing ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Male ; Memory ; drug effects ; Mice ; Neuroprotective Agents ; pharmacology

The aim of the present study was to investigate the effect of precursor brain-derived neurotrophic factor (proBDNF) on survival and neurite outgrowth of cultured rat spiral ganglion neurons (SGNs). Spiral ganglions (SG) were collected from postnatal day 5 Sprague Dawley (SD) rats, then enzymatically digested and suspended. Dissociated SGNs were plated on poly-D-lysine/laminin coated eight-well chamber plates and maintained at 37 °C for 4 h to promote the attachment of the neurons. Cultured SGNs were randomly divided into five groups: control group, BDNF group (BDNF 10 ng/mL), C10 group (proBDNF 10 ng/mL), C50 group (proBDNF 50 ng/mL), and C100 group (proBDNF 100 ng/mL). All groups were incubated in a serum-free medium. 48 h after incubation, SGNs were fixed and stained for βIII tubulin. Immunostaining of the cultured SGNs showed that, compared with the control group, the cellular survival of C50 group and C100 group were significantly reduced (P < 0.001). Furthermore, surviving numbers of the three proBDNF-treated groups were all lower than the BDNF group. In order to assess the effect of proBDNF on cell morphology, SGNs were divided into two categories: SGNs with or without neurites. The results demonstrated that proBDNF significantly increased the proportions of SGNs without neurites in C10, C50 and C100 groups compared with that in control group (P < 0.001). In addition, c-Jun N-terminal kinase (JNK) inhibitor, SP600125 (20 μmol/L) significantly increased the surviving number of SGNs in C50 group. These results suggest that proBDNF reduces the survival rate of cultured SGNs and inhibits the sprouting of neurites. Furthermore, the inhibition of JNK signaling attenuates the effect of proBDNF on SGNs survival.
Animals ; Axons ; physiology ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Survival ; Cells, Cultured ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; Neurites ; physiology ; Neurons ; cytology ; Protein Precursors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spiral Ganglion ; cytology