Intrahepatic Cholestasis of Pregnancy Increases Inflammatory Susceptibility in Neonatal Offspring by Modulating Gut Microbiota.

Intrahepatic cholestasis of pregnancy (ICP) is a liver disease of pregnancy that is characterized by increased bile acid levels in maternal serum. Studies have shown that cholestatic pregnancy can result in long-term metabolic disturbances in the offspring. However, how ICP shapes the offspring's immunity and predisposition to inflammatory disorders at an early stage is unknown. In this study, we investigated the effect of maternal cholestasis on neonatal offspring metabolism and immune function. We compared 71 neonates with ICP mothers and 63 neonates with healthy mothers and found that the incidence of jaundice and infection was significantly higher in ICP offspring. Maternal serum total bile acid level was associated with blood cell counts in full-term ICP offspring. In animal experiments, a compensatory activation of hepatic and ileal farnesoid X receptor (FXR) and altered gut microbiota in the first week were found in ICP offspring. We also investigated lipopolysaccharide (LPS)-induced inflammatory responses in neonatal rats and found that ICP offspring were more susceptible to inflammation. To understand the correlation between congenital abnormal FXR activation and tissue immunity dysregulation, we assessed the effects of the FXR agonist GW4064 and FXR antagonist E/Z-GS in ICP offspring after LPS exposure. The expression of several pro-inflammatory cytokines significantly decreased after treatment with E/Z-GS but increased after treatment with GW4064. Treatment with the probiotic Lactobacillus rhamnosus LRX01 that inhibits FXR expression in the ileum reduced susceptibility to LPS exposure in ICP offspring. The current study indicated that cholestatic pregnancy may increase the susceptibility of the offspring to inflammation by altering bile acid metabolism and gut microbiota at an early stage. We suggest that supplementation with Lactobacillus rhamnosus LRX01, which inhibits FXR expression in the ileum, may improve intestinal immunity in ICP offspring.

14-3-3 gene family in spotted sea bass (Lateolabrax maculatus): Genome-wide identification, phylogenetic analysis and expression profiles after salinity stress.

The tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation (14-3-3) proteins are a group of highly conserved homologous and heterologous proteins involved in a wild range of physiological processes, including the regulation of many molecular phenomena under different environmental salinities. In this study, we identified eleven 14-3-3 genes from the spotted sea bass (Lateolabrax maculatus) genome and transcriptomic databases and verified their identities by conducting phylogenetic, syntenic and gene structure analyses. The spotted sea bass 14-3-3 genes are highly conserved based on sequence alignment, conserved domains and motifs, and tertiary structural feature. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 14-3-3 genes in gill of spotted sea bass under normal physiological conditions indicated that the expression level of 14-3-3 zeta was the highest among tested genes, followed by 14-3-3 theta. Furthermore, expression profiles of 14-3-3 genes in gill tissue (in vivo and in vitro) indicated that the 14-3-3 zeta and 14-3-3 theta genes were significantly induced by different environmental salinities in spotted sea bass, suggesting their potential involvement in response to salinity challenge. Our findings may lay the foundation for future functional studies on the 14-3-3 gene family in euryhaline teleosts.

[Isolation, identification and lead adsorption study of lead-resistant Lactobacillus casei strains from feces of healthy newborns].

OBJECTIVE: To isolate and identify lead-resistant Lactobacillus casei strains with lead adsorption ability from the stool of healthy newborns as a new source of bacteria for developing lead-eliminating food products. METHODS: MRS was used to isolate lead-resistant bacteria from the feces of 30 healthy and full-term neonates. A phylogenetic tree was constructed based on the morphological characteristics and 16S rRNA sequences of the isolated bacteria. Physiological and biochemical characterizations of the bacteria were performed according to the Berger's Systematic Bacteriology Handbook, followed by antimicrobial susceptibility test and acid-tolerant bile salt test. The adsorption capacity of Pb2+ of the bacteria was determined by inductively coupled plasma-optical emission spectroscopy (ICP-OES). RESULTS: Three strains of Lactobacillus casei were isolated, which were resistant to penicillin and ceftriaxone and could tolerate the exposure to 500 mg/L Pb2+. Acid-tolerant bile salt test showed that the bacteria were resistant to culture in the presence of artificial gastric juice (pH 2.0) for 3 h, and their survival rate reached 62.5% following exposure to 0.3% bile salt for 8 h. The bacteria showed a Pb2+ adsorption rate of 90.4% at a low Pb2+ concentration (1 mg/L) and of 86.27% at a high Pb2+ concentration (50 mg/L). CONCLUSION: Three Lactobacillus casei strains lead adsorption ability were isolated from the feces of newborns. These bacterial strains provide a new solution to alleviate lead poisoning by probiotic dietary.

[Effect of Electroacupuncture Intervention on Expression of NR 2 B Subunit of NMDA Receptor in Amygdala during Morphine Withdrawal in Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the spatial learning-memory ability and the expression of NR 2 B subunit of NMDA receptor in the amygdala (AMG) in rats with morphine withdrawal. METHODS: A total of 40. SD rats were randomized into control, model, manual acupuncture and EA groups (n = 10 in each group). The morphine withdrawal model was established by subcutaneous injection of morphine hydrochloride injection at doses of 20, 30, 40, 50 and 50 mg . kg-1 . d-1 continuously for 5 days, followed by injection of naloxone (i. p., 3 mg/kg) for rapid induction of withdrawal syndrome. Manual acupuncture or EA stimulation was applied to bilateral "Shenshu"(BL 23) and "Zusanli" (ST 36), once daily for 6 days. Morris water maze swimming test was conducted for detecting the rats' spatial learning-memory ability. The expression levels of NR 2 B protein and mRNA in AMG was measured by Western blot and Real time-PCR, respectively. RESULTS: Compared to the control group, the escape latency on day 5 of swimming tests was obviously prolonged in the model group (P<0. 01), while in comparison with the model group, the escape latencies in both manual acupuncture and EA groups were obviously shortened (P<0. 01), suggesting an improvement of the rats' learning-memory ability. In addition, the expression levels of NR 2 B protein in both manual acupuncture and EA groups and that of NR 2 B mRNA in the EA group were significantly higher than those of the model group (P<0. 01, P<0. 05). CONCLUSION: Both manual acupuncture and EA interventions can improve the learning-memory ability in morphine withdrawal rats, which is probably partially related to their effects in up-regulating the expression of NR 2 B in the AMG.

Protection against Escherichia coli O157:H7 challenge by immunization of mice with purified Tir proteins.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 infections cause serious public health problems worldwide. The translocation intimin receptor (Tir) is responsible for adhesion and attaching and effacing lesions. In the current study, we used a mitomycin-treated mouse model to evaluate the efficacy of subcutaneous vs intranasal administration of the recombinant Tir as vaccine. Following immunization, mice were infected with E. coli O157:H7 and faces were monitored for shedding. Mice immunized intrasally with purified Tir proteins produced higher IgG and IgA titers in serum and feces, resulting in significant reductions in fecal shedding of EHEC O157 and higher a survival rate (92.9%), compared with subcutaneous or control immunizations. These results demonstrate the potential for the use of Tir proteins in mucosal vaccine formulations to prevent colonization and shedding of E. coli O157:H7. Therefore, purified Tir protects mice against EHEC challenge after intranasal immunization and is worth further clinical development as a vaccine candidate.

[Study on the allergens of shrimps and crabs].

AIM: To investigate the allergen composition in the species of shrimp and crab. METHODS: The main allergens of shrimps and crabs were analyzed by SDS-PAGE and Western blot assay. RESULTS: The molecule weight of main allergic protein in shrimps was near to 36 000, and the main allergic proteins in different kinds of crabs were about 29 000, 36 000, 66 000, 89 000 molecule weight. CONCLUSION: There are common allergens among different kinds of shrimps and crabs.

[Study on Rotavirus detection by nucleic acid sequence based amplification].

Rotavirus is one of the major cause of the viral gastroenteritis throughout the world. A nucleic acid sequence-based amplification(NASBA) technique for the detection of rotavirus in faecal specimens was developed and compared to the RT-PCR technique. Primers were designed according to the high conserved region of VP7 gene. Amplicons were detected by denaturating agarose gel, and then demonstrated by Northern hybridization with digoxigenin-labeled oligonucleotide probe. The anticipative specific amplification product is 392bp,and no nonspecific products appear even the concentration of nontarget nucleic acid as high as 1 microg/microL. A detection limit of 50 pg target RNA/mL is obtained when the optimal amplification time of 3h used.The NASBA assay will be a favourable alternative to RT-PCR for the investigation of rotavirus outbreaks as a routine diagnostic test in the near future because it is shown to be highly sensitive, specific and do not require specialized equipment.

[Study on rotavirus detection with single-tube seminested RT-PCR method in shellfish].

Being a foodborne virus, Rotavirus was often carried by shellfish. At the present, RT-PCR was the most effective method of Rotavirus detection in shellfish, but its sensitivity was low because of low levels of virus contamination and PCR inhibitors in shellfish. So contaminated shellfish experimentally in laboratory, and imitated the natural environment to concentrate Rotavirus, then detected by the developed single-tube seminested RT-PCR. Compared with ELISA and the conventional RT-PCR, the detection sensitivity is improved by 1000 times and 10 times respectively. In addition, the outer and inner contamination is reduced dramatically and the detection time is decreased from 6h to 4.5h. When the whole shellfish and only the digestive tissues of shellfish serve as samples, the detection ratio and sensitivity are more higher when sample is the latter.