Guilin Cheng

Effect of heat stress on the porcine small intestine: a morphological and gene expression study.

With the presence of global warming, the occurrence of extreme heat is becoming more common, especially during the summer, increasing pig susceptibility to severe heat stress. The aim of the current study was to investigate changes in morphology and gene expression in the pig small intestine in response to heat stress. Forty eight Chinese experimental mini pigs (Sus scrofa) were subjected to 40 degrees C for 5h each day for 10 successive days. Pigs were euthanized at 1, 3, 6, and 10 days after heat treatment and sections of the small intestine epithelial tissue were excised for morphological examination and microarray analyses. After heat treatment, the pig rectal temperature, the body surface temperature and serum cortisol levels were all significantly increased. The duodenum and jejunum displayed significant damage, most severe after 3 days of treatment. Microarray analysis found 93 genes to be up-regulated and 110 genes to be down-regulated in response to heat stress. Subsequent bioinformatic analysis (including gene ontology and KEGG pathway analysis) revealed the genes altered in response to heat stress related to unfolded protein, regulation of translation initiation, regulation of cell proliferation, cell migration and antioxidant regulation. Heat stress caused significant damage to the pig small intestine and altered gene expression in the pig jejunum. The results of the bioinformatic analysis from the present study will be beneficial to further investigate the underlying mechanisms involved in heat stress-induced damage in the pig small intestine.

Enhancement of Astragalus polysaccharide on the immune responses in pigs inoculated with foot-and-mouth disease virus vaccine

The effects of Astragalus polysaccharides (APS) on the immune response in pigs immunized with foot-and-mouth disease virus (FMDV) vaccine were investigated. Fifteen pigs were randomly divided into five groups. Four groups were vaccinated with a FMDV inactivated vaccine. Pigs in three experimental groups were administered varying doses of APS (APS1, 5mg/kg; APS2, 10mg/kg; APS3, 20mg/kg). The influence of APS on the number of CD3(+)CD4(-)CD8(+) cytotoxic T cells, CD3(+)CD4(+)CD8(+) T helper memory cells, and CD3(-)CD4(-)CD8(+) natural killer cells among peripheral blood lymphocytes (PBL) in the three APS groups were significant compared to the vaccine group. In vitro stimulation of PBL by Con A and LPS in APS groups induced a stronger proliferative response at 2 and 6 weeks post-inoculation (PI). APS markedly increased the titer of FMDV-specific antibody in a dose-dependent manner, and up-regulated mRNA expression of IFN-γ and IL-6. APS could potentially be used as an immunomodulator for a FMDV vaccine and provide better protection against FMDV.

Microarray Analysis of Gene Expression Profiles of rat Small Intestine in response to Heat Stress

Ambient temperature is a critical factor that affects biological organisms in many ways. In this study, the authors investigated gene expression changes in rat small intestine in response to heat stress. Male Sprague-Dawley rats were randomly divided into control and heat-stressed groups. Both groups were housed at 25 °C, although the heat-stressed group was also subjected to 40 °C for 2 h each day for 10 successive days. Rats were sacrificed 1, 3, 6, and 10 days after heat treatment, and sections of their small intestine epithelial tissue were excised for morphological examination and microarray analyses. The rat rectal and body surface temperatures and serum cortisol levels were all significantly increased after heat treatment (p < 0.05). The jejuna were significantly damaged by 3 days after heat treatment began. Microarray analysis showed that 422 genes were differentially expressed, of which 290 genes were significantly upregulated and 132 genes were significantly downregulated. Subsequent bioinformatics analyses revealed that the differentially expressed genes were mainly related to stress, immune regulation, and metabolism processes. The bioinformatics analysis of the differentially expressed genes should be beneficial to further investigations on the underlying mechanisms involved in heat stress–induced damage in the small intestine.

Taguchi approach for anti-heat stress prescription compatibility in mice spleen lymphocytes In Vitro

Heat stress (HS) may induce immunosuppression as well as inhibit the proliferation of lymphocytes. This study evaluated the effects on immune function of our prescription on splenic lymphocytes under HS as well as its compatibility. The effects of four herbal extracts from Agastache rugosa, Atractylodes lancea, Cortex Phellodendri, and Gypsum Fibrosum on heat treated splenic lymphocytes were investigated and the compatibility of the prescription was also explored by using the Taguchi method. This study revealed changes in proliferation by traditional Chinese medicines of splenic lymphocytes after HS. Proliferation in the HS group was significantly lower than the control group. Under HS, the effects of higher concentrations of Agastache rugosa (100 and 200 μg/mL), Atractylodes lancea (100 and 200 μg/mL), Cortex Phellodendri (50 and 100 μg/mL) and Gypsum Fibrosum (100 and 200 μg/mL) caused a significant increase on ConA/LPS-induced proliferation of lymphocytes than lower concentrations. We, therefore, conclude that the prescription of traditional Chinese medicines may recover splenic lymphocytes from the immunosuppression induced by HS. The Taguchi design, which allows rapid and high efficiency for the selection of the best conditions for our prescription on HS-treated splenic lymphocytes, demonstrated that Agastache rugosa (200 μg/mL), Atractylodes lancea (200 μg/mL), Cortex Phellodendri (100 μg/mL) and Gypsum Fibrosum (100 μg/mL) were the optimal conditions for the prescription. The validation experiment confirmed that our composition in optimum extraction conditions enhanced effects on ConA or LPS-stimulated lymphocytes under HS. The results showed that the Taguchi optimization approach is a suitable method for optimization of the composition of prescription.

Heat Stress Induced Apoptosis in Rat Intestinal Epithelial Cell Line-6 (IEC-6)

The aim of this paper is to study the effect of heat stress on the apoptosis of rat intestinal epithelial cell line-6 (IEC-6), using fluorescent dyeing technology to observe its morphological changes, flow cytometry to detect the apoptosis cells rate, RT-PCR to measure the apoptosis relevant genes. The results showed that there were evident morphological changes in the IEC-6 treated by the heat stress (42 °C 4h) and the cell membrane permeability was significantly enhanced. The apoptosis cells rate of early and late stages were significantly increased (p < 0.05). RT-PCR measurement revealed that the apoptosis promoting gene BAX expression was were significantly increased (p < 0.05) and the apoptosis inhibiting gene bcl-2 expression was were significantly decreased (p < 0.05). Meanwhile, the caspase-3 gene expression was were significantly increased (p < 0.05). This preliminary result showed the heat stress can induce the apoptosis of IEC-6.

Effect of Spleen Deficiency Syndrome on the Expression of TLR2 and TLR4 in Rats

[Objective]The aim of the paper was to study the expression of TLR2 and TLR4 in the rat’s spleen deficiency syndrome (SDS) induced by reserpine. [Method]Twelve SD rats were divided into the control group and SDS group. After 7 days, the rats were sacrificed. The duodenum, jejunum and ileum was removed and washed in phosphatebuffered saline (PBS). One part of each intestine sections was fixed and paraffin embedded. The TLR2 secretion were observed by immunohistochemical staining. The other part was to measure the expressions of TLR2 and TLR4 were measured using RT-PCR. [Result]The results indicated that comparing with control group, the TLR2 secretion of SDS group was significantly decreased. The expression of TLR2 and TLR4 of SDS group in jejunum were significantly decreased (P < 0. 01), but TLR4 in ileum was increased (P < 0. 01). [Conclusion]In conclusion, these results indicated that SDS induced damage in jejunum, which resulted in the down-regulation of the expression of TLR2 and TLR4.

Effect of Huchuanqing on chicken infectious bronchitis

In order to study the effect of the traditional Chinese medicine “Huchuanqing Decoction” in preventing and curing the chicken infectious bronchitis disease, a chicken infectious bronchitis model was established by artificial infecting chicken infectious bronchitis virus. This paper studied the effect of virus on the chicken production performance, the changes of cytokine IFN-γ, IL-6 and TNF-α concentration in chicken serums and the effect of Huchuanqing Oral Liquid on it. The results provided evaluation indexes and the basis in developing Chinese medicines for preventing and curing the chicken infectious bronchitis.

Clinical trial report of treating E. coli. infected chicken by Changliqing decoction

E. colibacillosis is considered to be one of the major bacterial diseases in the poultry industry. In this study, E. colibacillosis was induced by artificial infection in chicken and cured by Changliqing decoction, thus to verify the therapeutic effect via clinical trials. Several factors, including clinical symptoms, mortality rate, hatching performance, autopsy changes, were monitored and analyzed. Data showed that compared with positive control, the mortality rate were dramatically lower, indicating a significance of difference (P&#60;0.05), but are dosage independent. Furthermore, high and medium dosage groups of Changliqing decoction treated chicken showed remarkably higher rate on daily gain against both positive control and positive drug control (P&#60;0.05). Additionally, by analyzing the clinical symptoms and autopsy changes, there is no significant difference (P>0.05) between the control group and Changliqing decoction treated groups (dosage irrelevant). All above results indicates that Changliqing decoction treatment potentially performs positive for curing E. colibacillosis on clinical level and recommended dosage is about 1.5 ml/kg once a day.