In vaccination,traditional vaccines are administered intramuscularly or subcutaneously,mainly inducing humoral immunity,with weak induction of mucosal immunity and insufficient protection against respiratory and gastrointestinal viruses.Current research has found that immuni-zation vaccines can effectively induce mucosal immunity through oral or nasal administration,but there is a lack of effective mucosal adjuvants,and there is an urgent need for the development of safe and effective new mucosal adjuvants.With the advancement of nano-adjuvant research,novel nanoparticles have attracted widespread attention due to their good mucosal adhesion,efficient an-tigen-loading ability,biocompatibility,and advantages in immune-enhancement,nano-size effect,tissue-targeting,and slow release of antigens,etc.,and are expected to be used as novel mucosal ad-juvants in the prevention and treatment of animal diseases.To date,significant progress has been made in research targeting mucosal adjuvants such as polymeric nanoparticles,inorganic nanoparti-cles,and liposomes.This paper reviews the application of nanomucosal adjuvants in animal vaccines and their mechanisms of action at home and abroad,and analyzes the physicochemical factors af-fecting the immune-enhancing effect of nanoparticle adjuvants,with a view to providing a reference for the development of novel mucosal adjuvants.

In vaccination,traditional vaccines are administered intramuscularly or subcutaneously,mainly inducing humoral immunity,with weak induction of mucosal immunity and insufficient protection against respiratory and gastrointestinal viruses.Current research has found that immuni-zation vaccines can effectively induce mucosal immunity through oral or nasal administration,but there is a lack of effective mucosal adjuvants,and there is an urgent need for the development of safe and effective new mucosal adjuvants.With the advancement of nano-adjuvant research,novel nanoparticles have attracted widespread attention due to their good mucosal adhesion,efficient an-tigen-loading ability,biocompatibility,and advantages in immune-enhancement,nano-size effect,tissue-targeting,and slow release of antigens,etc.,and are expected to be used as novel mucosal ad-juvants in the prevention and treatment of animal diseases.To date,significant progress has been made in research targeting mucosal adjuvants such as polymeric nanoparticles,inorganic nanoparti-cles,and liposomes.This paper reviews the application of nanomucosal adjuvants in animal vaccines and their mechanisms of action at home and abroad,and analyzes the physicochemical factors af-fecting the immune-enhancing effect of nanoparticle adjuvants,with a view to providing a reference for the development of novel mucosal adjuvants.

ObjectiveTo clarify the relationship between intestinal flora and intestinal motility in rats with slow transit constipation （STC） and qi stagnation syndrome by conducting a pseudo-sterile experiment and fecal microbiota transplantation （FMT） technology. MethodsTwenty-four Wistar rats were randomly divided into normal group （n=6）， STC with qi stagnation pattern group （n=6） and pseudo-sterile group （n=12）. In the STC group with qi stagnation pattern， 3 mg/kg of loperamide suspension by intragastric administration combined with tail clamping stimulation were performed to establish the rat model of STC with qi stagnation pattern. After successful modeling， fresh feces from the rats in the STC with qi stagnation pattern group and the normal group were collected to prepare 100 mg/ml of fecal bacterial suspension. In the pseudo-sterile group， the antibiotic cocktail method was used （a mixed antibiotic suspension containing bacitracin， streptomycin sulfate， and neomycin sulfate at 20 mg/ml each was administered intragastrically） to establish pseudo-sterile rats model. After successful modeling， the rats were randomly divided into normal fecal bacterial liquid group and STC with qi stagnation pattern fecal bacterial liquid group， with six rats in each group， and then were given 10 ml/kg of the prepared corresponding rat fecal bacterial suspension by gavage. Rats in STC with qi stagnation pattern group were given an equal volume of sterile water by gavage. All groups were administered once a day for 7 consecutive days. The small intestinal propulsion rate of the STC with qi stagnation pattern group， the normal fecal bacterial liquid group， and STC with qi stagnation pattern fecal bacterial liquid group were compared. ELISA method was used to detect serum 5-hydroxytryptamine （5-HT） levels. Immunohistochemistry was used to detect the positive expression levels of 5-hydroxytryptamine 3 receptor （5-HT3R） and 5-hydroxytryptamine 4 receptor （5-HT4R） in colon tissue. Western blot method was used to detect the protein expression levels of tryptophan hydroxylase 1 （TPH1）， tryptophan hydroxylase 2 （TPH2）， serotonin transporter （SERT）， and monoamine oxidase A （MAO-A） in colon tissue. ResultsCompared to those in the normal fecal bacterial liquid group， the small intestinal propulsion rate， serum 5-HT level， positive expression of 5-HT3R and 5-HT4R in colon tissue， and protein expression of TPH1， TPH2， SERT and MAO-A significantly decreased in the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＜0.05）. There was no statistically significant difference in the indicators between the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＞0.05）. ConclusionThe intestinal flora in STC rats with qi stagnation pattern can lead to a slowdown in intestinal transmission function， whose mechanism may be related to intestinal motility disorders affected by the synthesis， transport， metabolism and other pathways of 5-HT.

Objective:To investigate the characteristics of anorectal dynamics in elderly patients with functional defecation disorders(FDD), and to provide a basis for their diagnosis, treatment and prevention.Methods:In this retrospective study, 226 patients with FDD receiving 3D high-resolution anorectal manometry were divided into an elderly group(93 cases)and a non-elderly group(133 cases). Results from anorectal manometry parameters were compared and analysis of patterns of anorectal pressure changes in elderly participants based on sex, the Bristol stool classification and clinical symptoms was conducted.Results:The resting anal pressure, rectal pressure and anal relaxation rate were lower( t=-3.407, -2.051, Z=2.548, P=0.001, 0.040, 0.011)and the volume of first sensation was higher( t=1.998, P=0.047)in the elderly group than in the non-elderly group.The maximum anal squeezing pressure, residual anal pressure and maximum tolerated volume were higher( t=4.589, 4.730, 2.025, all P<0.05), whereas the anal relaxation rate and anorectal pressure gradient were lower in elderly men than in elderly women( Z=4.059, t=-3.714, P<0.001 for both). Regarding the types of FDD, both the elderly group and the non-elderly group were dominated with type Ⅱ defecation disorder, with more men than women having type Ⅱ defecation disorder in the elderly group( χ2=10.343, P=0.001). In cases of paradoxical sphincter contraction during simulated defecation, the incidence in the elderly group was 80.65%(75/93), which was higher than 68.42%(91/133)in the non-elderly group( χ2=4.194, P=0.041). The volume of first sensation, volume of first defecation sensation, and maximum tolerated volume of patients in the elderly group without the urge to defecate were(59.86±23.84)ml, (96.76±34.61)ml, and(144.32±30.57)ml, respectively, higher than those of patients with the urge to defecate(46.79±17.20)ml, (75.26±28.75)ml, and(120.00±40.28)ml( t=-2.241, -2.493, -2.891, P=0.027, 0.014, 0.005). The rectal pressure(26.52±16.08)mmHg of patients with defecation dyssynergia was lower than that of patients without defecation dyssynergia(39.91±8.82)mmHg(1 mmHg=0.133 kPa)( t=-3.128, P=0.002), while the resting anal pressure of patients with defecation dyssynergia(90.60±28.44)mmHg was higher than that of patients without defecation dyssynergia(73.65±27.10)mmHg( t=-2.201, P=0.030). The resting anal pressure and maximum anal squeezing pressure in patients with anal blockage sensation[(87.11±24.64)mmHg, (149.28±48.29)mmHg]were higher than those in patients without anal blockage sensation[(72.43±20.02)mmHg, (121.76±26.35)mmHg]( t=2.954、3.066, P=0.004、0.003). There was no significant difference in values from parameters of anorectal dynamics between patients with different Bristol stool types, with and without incomplete defecation or with different degrees of abdominal distension(all P>0.05). Conclusions:Anorectal dynamics in patients with FDD are characterized by paradoxical anal sphincter movements, but older patients with FDD are mainly characterized by inadequate rectal propulsion, pelvic floor muscle dysfunction and reduced rectal sensitivity.

OBJECTIVE: To investigate the changes in the expression of voltage-gated potassium channel subunit KCNA2 in the dorsal root ganglion (DRG) neurons of rats with osteoarthritis (OA) pain induced by sodium monoiodoacetate and explore the mechanism. METHODS: A total of 156 adult male Sprague-Dawley rats were randomly divided into blank control group, saline group and intra-articular monoiodoacetate injection-induced OA group. The paw withdrawal mechanical threshold (PWMT) was measured before and at 1, 2, 4, and 6 weeks after monoiodoacetate injection. At 4 weeks after the injection, the pathological changes in the knee joints were analyzed using HE staining and Safranin O-Fast Green staining, and the expression of activating transcription factor 3 (ATF-3) and inducible nitric oxide synthase (iNOS) in the DRG neurons were detected by immunofluorescence staining. The expression of mRNA in the DRG neurons was detected by RT-qPCR at 1, 2, 4 and 6 weeks after the injection. The expression of KCNA2 in the DRG was measured by Western blotting, and the methylation level of promoter region was measured by MSPCR at 4 weeks after the injection. RESULTS: The PWMT of the rats in OA group was significantly decreased at 2, 4, and 6 weeks after the injection as compared with the baseline ( < 0.05 or < 0.001) as well as the control group ( < 0.05 or < 0.001). Four weeks after the intra-articular injection, fractures and defects on the surface of the articular cartilage, bone hyperplasia, and blurred tidal line were observed in the rats in OA group, but no obvious pathological changes were detected in the control or saline groups. Compared with those in the control group, the expressions of ATF-3 and iNOS were significantly increased ( < 0.01) at 4 weeks after injection; the expression of mRNA at 2, 4 and 6 weeks and the expression of KCNA2 protein at 4 weeks were all significantly decreased ( < 0.05 or < 0.01), and the methylation level of gene was significantly increased at 4 weeks after the injection in OA group ( < 0.01). CONCLUSIONS The expression of KCNA2 is decreased in the DRG neurons of rats with OA pain likely as a result of enhanced methylation of promoter region.
Animals ; Disease Models, Animal ; Ganglia, Spinal ; Knee Joint ; Kv1.2 Potassium Channel ; metabolism ; Male ; Osteoarthritis ; complications ; metabolism ; Pain ; etiology ; metabolism ; Promoter Regions, Genetic ; Rats ; Rats, Sprague-Dawley

The present study was aimed to evaluate resting-state functional connectivity and topological properties of brain networks in narcolepsy patients compared with healthy controls. Resting-state fMRI was performed in 26 adult narcolepsy patients and 30 matched healthy controls. MRI data were first analyzed by group independent component analysis, then a graph theoretical method was applied to evaluate the topological properties in the whole brain. Small-world network parameters and nodal topological properties were measured. Altered topological properties in brain areas between groups were selected as region-of-interest seeds, then the functional connectivity among these seeds was compared between groups. Partial correlation analysis was performed to evaluate the relationship between the severity of sleepiness and functional connectivity or topological properties in the narcolepsy patients. Twenty-one independent components out of 48 were obtained. Compared with healthy controls, the narcolepsy patients exhibited significantly decreased functional connectivity within the executive and salience networks, along with increased functional connectivity in the bilateral frontal lobes within the executive network. There were no differences in small-world network properties between patients and controls. The altered brain areas in nodal topological properties between groups were mainly in the inferior frontal cortex, basal ganglia, anterior cingulate, sensory cortex, supplementary motor cortex, and visual cortex. In the partial correlation analysis, nodal topological properties in the putamen, anterior cingulate, and sensory cortex as well as functional connectivity between these regions were correlated with the severity of sleepiness (sleep latency, REM sleep latency, and Epworth sleepiness score) among narcolepsy patients. Altered connectivity within the executive and salience networks was found in narcolepsy patients. Functional connection changes between the left frontal cortex and left caudate nucleus may be one of the parameters describing the severity of narcolepsy. Changes in the nodal topological properties in the left putamen and left posterior cingulate, changes in functional connectivity between the left supplementary motor area and right occipital as well as in functional connectivity between the left anterior cingulate gyrus and bilateral postcentral gyrus can be considered as a specific indicator for evaluating the severity of narcolepsy.

Objective To evaluate the role of different opioid receptors in morphine postconditioning-induced reduction of ischemia-reperfusion (Ⅰ/R) injury in isolated rat hearts.Methods Healthy adult male Sprague-Dawley rats,aged 7-8 weeks,weighing 250-300 g,were anesthetized with chloral hydrate.The hearts were removed and retrogradely perfused with oxygenated K-H solution in a Langendorff apparatus.Forty-eight Langendorff-perfused rat hearts were divided into 4 groups (n=12 each) by a random number table method:Ⅰ/R group,morphine postconditioning group (M group),δ receptor antagonist naltrexone plus morphine postconditioning group (N+M group) and κ receptor antagonist nor-binaltorphimine plus morphine postconditioning group (B+M group).Hearts were subjected to 4 cycles of perfusion with K-H solution containing 1 μmol/L morphine for 15 s and then with K-H solution containing no morphine for 15 s in group M.In N+M group and B+M group,hearts were perfused with 5 μmol/L naltrindole and 5 μmol/L nor-binaltorphimine,respectively,starting from 10 nin of equilibration until 5 min of reperfusion,and morphine postconditioning was similar to those previously described in group M.Heart rate and the maximum rate of increase or decrease in left ventricular pressure (±dp/dtmax) were recorded at 20 min of equilibration and 30 and 60 nin of reperfusion.Corona~ effluent was collected at 20 min of equilibration and 60 min of reperfusion for measurement of the activity of creatine kinase by colorimetric assay.Eight hearts were obtained at 60 min of reperfusion for determination of myocardial infarct size.Four hearts were obtained at 60 min of reperfusion for detection of the expression of microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ) and LC3 Ⅱ by Western blot.LC3 Ⅱ/LC3 Ⅰ ratio was calculated.Results Compared with the baseline value at 20 min of equilibration,heart rate and +dp/dt were significantly decreased at 30 and 60 min of reperfusion,and the activity of creatine kinase in coronary effluent was increased at 60 min of reperfusion in four groups (P＜0.05).Compared with group Ⅰ/R,the activity of creatine kinase in coronary effluent,percentage of myocardial infarct size and LC3 Ⅱ/LC3 Ⅰ ratio were significantly decreased in group M (P＜0.05).Compared with group M,the activity of creatine kinase in coronary effluent,percentage of myocardial infarct size and LC3 Ⅱ/LC3 Ⅰ ratio were significantly increased in N+M and B+M groups (P＜0.05).Conclusion δ and κ opioid receptors are involved in morphine postconditioning-induced reduction of Ⅰ/R injury in isolated rat hearts,and the mechanism may be related to inhibiting the level of autophagy.

Objective To explore the risk,diagnosis,treatment and prognosis of newborns suffering from congenital latent tuberculosis infection (CLTBI) by vertical transfer of mycobacterium tuberculosis(VTRTB) from mothers with active tuberculosis (TB) mothers during pregnancy.Methods The clinical data of 7 children with CLTBI follow-up of 1-6 years in Children's Hospital of Nanjing Medical University and 2 cases from literatures were reviewed,and they were divided into simple CLTBI group (n =5) and active TB group (n =4).Combined with the maternal history,the epidemiology,clinical features,treatment,prognosis and relative factors of the children were analyzed.Results Eight mothers had active TB during pregnancy,and premature delivery rate and low birth weight rate were 55.56％ (5/9 cases) respectively.The positive conversion rates of purified protein derivative tests (PPD),acid-fast bacilli (AFB) and TB culture were 6/9 cases,0/9 cases and 1/9 cases,respectively.Positive results of T cell enzyme-linked immunospot tuberculous test (T-SPOT.TB) and TB-DNA by fluorescent quantitative polymerase chain reaction (FQ-TB-DNA) were detected in 4 cases respectively,1 of 3 patients with T-SPOT.TB and 1 of 3 patients with FQ-TB-DNA were positive.There were 5 cases with chemoprophylaxis of anti-TB drugs for 1-6 years with isonicid or rifampicin;3 cases with pulmonary hilar or mediastinal lymph nodes enlargement,and 3 cases with pulmonary diffuse punctate,nodular or patchy high density shadow in 3 cases;the rate of VTRTB in 9 cases was 8/9 (88.89％) based on laboratory examination of etiology and immunology,and the risk of active TB in infants and early childhood was close to 50％ in neonates with CLTBI;the incidence of active TB was reduced from 100％ (4/4 cases) of non-intervention down to 0 (0/5 case) of intervention.Conclusions Etiology,immunology,imaging examination of TB and clinical efficacy,can prompt active TB and retrospective diagnosis of CLTBI,and application of T-SPOT.TB and FQ-TB-DNA for early diagnosis of CLTBI has a higher applied value;prophylactic and treatment of anti-tuberculosis drugs can reduce the progression of CLTBI to active TB and improve outcomes.

Objective To evaluate the effect of dexmedetomidine on the intestinal mucosal injury in the patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Forty patients of both sexes with rheumatic heart disease,aged 32-64 yr,weighing 40-75 kg,of ASA physical status Ⅱ or Ⅲ (NYHA class Ⅱ or Ⅲ),scheduled for elective cardiac valve replacement with CPB,were randomly divided into 2 groups (n =20 each) using a random number table:control group (group C) and dexmedetomidine group (group D).After induction of anesthesia,the patients were endotracheally intubated and mechanically ventilated.Anesthesia was maintained with 0.8％-2.0％ sevoflurane inhalation and intermittent iv boluses of sufentanil 0.5-1.0 μg/kg and vecuronium 0.04-0.06 mg/kg.Before routine induction of anesthesia,a loading dose of dexmedetomidine 1 μg/kg was injected intravenously over 10 min,followed by continuous infusion at 0.3 μg · kg-1 · h-1 until the end of surgery in group D,while the equal volume of normal saline was given in group C.Before CPB,at 30 min after aortic clamping,at the termination of CPB,at the end of surgery and at 6 and 24 h after surgery,central venous blood samples were taken for determination of concentrations of tumor necrosis factor-alpha,interleukin-6 (IL-6) and IL-10 and intestinal fatty acid binding protein in plasma (by ELISA),and the plasma concentration of endotoxin (using turbidimetry).The time of postoperative mechanical ventilation and duration of ICU stay were recorded.Results Compared with group C,the concentrations of tumor necrosis factor-alpha,IL-6,IL-10 and endotoxin and intestinal fatty acid binding protein in plasma were significantly decreased,and the time of postoperative mechanical ventilation and duration of ICU stay were shortened in group D.Conclusion Dexmedetomidine infused continuously at 0.3 μg · kg-1 · h-1 (until the end of surgery) after a loading dose of 1 μg/kg before routine induction of anesthesia can reduce intestinal mucosal injury in the patients undergoing cardiac valve replacement with CPB.