This study aims to investigate the effects of Pulsatilla decoction(PD)on the 5-HT sig-naling system in the hippocampus of damp-heat diarrhoea(DHD)rats.Twenty-four male SD rats were randomly divided into four groups including the blank group,the model group,the PD group and the self-healing group.Except for the blank group,the rats in each group were induced by"high sugar and high fat,high temperature and high humidity,and E.coli poisoning"to establish a of rat model DHD,and were treated by gavage with PD.Changes in body weight,temperature,food intake and water intake,routine blood tests and histopathological changes in the colon were recor-ded to comprehensively determine the modelling condition of rats with DHD;histopathological changes in the hippocampus of rats were observed,and real-time fluorescence PCR was used to de-termine the expression of IL-1β3,IL-6,TNF-α,IFN-γ and TPH1 mRNA in the hippocampus;West-ern blot and immunohistochemistry were used to detect the protein expression of tryptophan hydroxylase 1(TPH1),receptors(5-HT3R,5-HT4R,5-HT7R)in 5-HT signaling pathway in the hippocampus.The results showed that:PD significantly regulated the abnormal changes of body weight,food and water intake and blood routine indexes in rats with DHD,and significantly im-proved the pathological damage of colonic tissues;PD significantly lowered the expression of in-flammatory cytokines IL-1β,IL-6,IFN-γ,and TNF-α in hippocampus of rats with DHD(P＜0.05),and significantly reduced the expression of TPH1 mRNA in hippocampus of rats with DHD(P＜0.05).PD could increased the expression of 5-HT4R and 5-HT7R in the hippocampus of rats with DHD;reduced the expression of 5-HT3R and TPH in the hippocampus,among which 5-HT3R expression was significantly reduced.This study suggests that PD can affect the function of hippocampus in rats with DHD by regulating the 5-HT signaling pathway.

This study aims to investigate the effects of Pulsatilla powder(PP)on colonic intestinal flora and tryptophan metabolism in piglets with diarrhea.Twenty-four weaned piglets were ran-domly divided into the normal control group,model group,self-healing group,and PP group.The model of piglet diarrhea was established by the compound factor of external evil+internal injury+epi-demic virus,and treated with PP.During the modeling period,the body weight,mental status and fecal patterns of piglets were monitored daily.After the experiment,the colon tissues were collect-ed for histopathological observation,and the inflammatory factors IL-1β and IL-6 in the colon tis-sue was detected by ELISA,16S rRNA was used to analyze colonic intestinal flora and RT-qPCR was used to determine the expression of genes related to tryptophan metabolism in the colon.The results showed that,compared with the model group,PP significantly improved the pathological status of the colon in piglets with diarrhea,with elevated body mass and a significant increase in goblet cells(P＜0.05);and significantly reduced the levels of IL-1β and IL-6 in the colon of diar-rhea piglets(P＜0.05).The results of intestinal flora showed that PP increased the abundance of Firmicutes(P＜0.01),Spirochaeota(P＜0.01)and Lactobacillus(P＜0.05),reduced the abun-dance of Bacteroidota(P＜0.05)、Prevotella_NK3B31_group(P＜0.05)and Clostridium_sensu_stricto 1(P＜0.01).Meanwhile,PP significantly reduced the expression level of TPH1 mRNA(P＜0.05),increased the expression levels of AhR(P＜0.05)and IL-22 mRNA in the co-lon of diarrhea piglets.These results indicates that PP may alleviate diarrhea in piglets by regula-ting colonic intestinal flora and genes related to intestinal tryptophan metabolism.This study pro-vides data support for the subsequent investigation of piglet diarrhea prevention and treatment.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

This study aims to investigate the effects of Pulsatilla decoction(PD)on the 5-HT sig-naling system in the hippocampus of damp-heat diarrhoea(DHD)rats.Twenty-four male SD rats were randomly divided into four groups including the blank group,the model group,the PD group and the self-healing group.Except for the blank group,the rats in each group were induced by"high sugar and high fat,high temperature and high humidity,and E.coli poisoning"to establish a of rat model DHD,and were treated by gavage with PD.Changes in body weight,temperature,food intake and water intake,routine blood tests and histopathological changes in the colon were recor-ded to comprehensively determine the modelling condition of rats with DHD;histopathological changes in the hippocampus of rats were observed,and real-time fluorescence PCR was used to de-termine the expression of IL-1β3,IL-6,TNF-α,IFN-γ and TPH1 mRNA in the hippocampus;West-ern blot and immunohistochemistry were used to detect the protein expression of tryptophan hydroxylase 1(TPH1),receptors(5-HT3R,5-HT4R,5-HT7R)in 5-HT signaling pathway in the hippocampus.The results showed that:PD significantly regulated the abnormal changes of body weight,food and water intake and blood routine indexes in rats with DHD,and significantly im-proved the pathological damage of colonic tissues;PD significantly lowered the expression of in-flammatory cytokines IL-1β,IL-6,IFN-γ,and TNF-α in hippocampus of rats with DHD(P＜0.05),and significantly reduced the expression of TPH1 mRNA in hippocampus of rats with DHD(P＜0.05).PD could increased the expression of 5-HT4R and 5-HT7R in the hippocampus of rats with DHD;reduced the expression of 5-HT3R and TPH in the hippocampus,among which 5-HT3R expression was significantly reduced.This study suggests that PD can affect the function of hippocampus in rats with DHD by regulating the 5-HT signaling pathway.

This study aims to investigate the effects of Pulsatilla powder(PP)on colonic intestinal flora and tryptophan metabolism in piglets with diarrhea.Twenty-four weaned piglets were ran-domly divided into the normal control group,model group,self-healing group,and PP group.The model of piglet diarrhea was established by the compound factor of external evil+internal injury+epi-demic virus,and treated with PP.During the modeling period,the body weight,mental status and fecal patterns of piglets were monitored daily.After the experiment,the colon tissues were collect-ed for histopathological observation,and the inflammatory factors IL-1β and IL-6 in the colon tis-sue was detected by ELISA,16S rRNA was used to analyze colonic intestinal flora and RT-qPCR was used to determine the expression of genes related to tryptophan metabolism in the colon.The results showed that,compared with the model group,PP significantly improved the pathological status of the colon in piglets with diarrhea,with elevated body mass and a significant increase in goblet cells(P＜0.05);and significantly reduced the levels of IL-1β and IL-6 in the colon of diar-rhea piglets(P＜0.05).The results of intestinal flora showed that PP increased the abundance of Firmicutes(P＜0.01),Spirochaeota(P＜0.01)and Lactobacillus(P＜0.05),reduced the abun-dance of Bacteroidota(P＜0.05)、Prevotella_NK3B31_group(P＜0.05)and Clostridium_sensu_stricto 1(P＜0.01).Meanwhile,PP significantly reduced the expression level of TPH1 mRNA(P＜0.05),increased the expression levels of AhR(P＜0.05)and IL-22 mRNA in the co-lon of diarrhea piglets.These results indicates that PP may alleviate diarrhea in piglets by regula-ting colonic intestinal flora and genes related to intestinal tryptophan metabolism.This study pro-vides data support for the subsequent investigation of piglet diarrhea prevention and treatment.

Objective To analyze the effects of different supplements on anemia related indexes in rural children. Methods A stratified method was adopted, and six villages (towns) in and around Qinghai Province were selected as intervention sits for the present study. A total of 304 children from 2 to 6 years old at each intervention site meeting the inclusion criteria were screened and divided into three groups (A, B, and C), who were intervened for 3 months. Serum vitamin A, vitamin D and hemoglobin levels were measured before and after the intervention. Results The hemoglobin level of 304 children before intervention was (118.65±16.07) g /L, and the prevalence of anemia was 9.54%. The vitamin A value, vitamin D value and hemoglobin value were increased after three months of the intervention. The changes of vitamin A value, vitamin D value and hemoglobin value in rural children in group C were significantly higher than those in groups A and B. The increase in vitamin A value in rural children aged 3 years was significantly higher than that in other age groups, and the increase in hemoglobin in rural children of 1 year old was significantly higher than that in other age groups. The increase in vitamin A value of rural children of other ethnic groups (mainly Tibetans) was significantly higher than that of Han and Hui nationalities, and the increase of hemoglobin value in Hui rural children was significantly higher than that in Han and other ethnic groups. Conclusion Vitamin A combined with iron dextran tablets was effective in preventing anemia in rural children.

Objective To investigate the functional and ultrastructural changes of cerebral microvascular endothelial cells at early stage following traumatic brain injury (TBI) in rats. Methods The rat models with closed brain injury were established with the improved Marmarous method. The expressions of thrombomodulin (TM) and von Willebrand factor (vWF) were determined by immunohistochemical techniques (5 rats per group) and real-time quantitative RT-PCR (5 rats per group) respectively at 1, 4, 8, 12, 24 hours and at days 3 and 7 after injury. Results TM and vWF started expression at 4 hours, reached peak at 24 hours and recovered to normal at day 7 after TBI. The expression levels of TM and vWF at different time points in sham control group showed statistical difference compared with damage group (P < 0.05). Conclusion The activation of the cerebral microvascular endothelial cells at early stage after TBI is the main mechanism of early secondary brain injury.

Objective To compare the effects of oxygen therapy and local pressurization in alleviating plateau hypoxia at high altitude.Methods Forty-five healthy male soldiers were investigated at an altitude of 3992 meters.The subjects were randomly divided into three groups, ie.an oxygen inhalation group, a single-soldier oxygen increasing respirator (SOIR) group and a BiPAP group.The oxygen inhalation group was treated with oxygen inhalation via nasal catheter at 2 L/min.SOIR was used to assist breath in the SOIR group.The BiPAP group were treated with bi-level positive airway pressure ventilation, with IPAP of 10 cm H20 and EPAP of 4 cm H2O.PaO2、PaCO2、SpO2 and heart rate were measured before and 30 minutes after the treatment.Results There were continuous increase of PaO2 from (53.30±4.88) mm Hg to (58.58±5.05) mm Hg and (54.43±3.01) mm Hg to (91.36±10.99) mm Hg after BiPAP ventilation and oxygen inhalation, respectively (both P < 0.01).However, the PaO2、of the SOIR group was decreased from (56.00±5.75) mm Hg to (50.82±5.40) mm Hg (P < 0.05).In the other hand, the PaCO2、 was increased from (30.41±1.51) mm Hg to (32.5±2.98) mm Hg in the oxygen inhalation group (P< 0.05), declined from (28.74±2.91) mm Hg to (25.82±4.35) mm Hg in the BiPAP group (P < 0.05), and didn't change significantly from (28.65±2.78)mm Hg to (29.75±3.89) nun Hg in the SOIR group (P > 0.05).Conclusions Both BiPAP ventilation and oxygen inhalation can alleviate plateau hypoxia by improving PaO2 at 3992 meter altitude while SOIR has no significant effect.

Objective To explore the role of the sensory neuropeptide in the chronic cough induced by postnasal drip syndrome(PNDs).Methods Patients of PNDs with and without chronic cough who came to the First Affiliated Hospital of Ji Nan University between Mar.2004 and Sep.2005 were enrolled and induced sputum by hypertonic saline aerosol inhalation was performed.Twenty-three cases of PNDs with chronic cough and 16 cases of PNDs without chronic cough were collected.Substance P(SP)and calcitonin gene-related peptide(CGRP)in the supernatant of the induced sputum were measured by radioimmunoassay.Results SP and CGRP in the supernatant of the induced sputum were significantly higher in the patients with chronic cough induced by PNDs[SP(345.14?72.58)mg/L,CGRP(573.78?210.96)mg/L]as compared with those normal subjects[SP(168.14?56.97)mg/L,CGRP(227.69?70.84)mg/L,P