Objective To evaluate the role of spinal neuronal Mas-related gene receptor C (MrgC) in the maintenance of bone cancer pain (BCP) in mice.Methods A total of 132 SPF male C3H/HeJ mice, aged 8-10 weeks, weighing 18-22 g, were randomly divided into 4 groups (n=33 each) using a random number table: sham operation group (S group) , BCP group, bovine adrenal medulla peptide 8-22 (BAM8-22, a highly selective MrgC agonist) group (group BAM), and MrgC antibody group (group MA).BCP was produced by injecting α-MEM 20 μl containing 2×105NCTC2472 cells into the distal medullary cavity of right femur bone.While α-MEM 20 μl was injected only in group S.The artificial cerebrospinal fluid 5 μl was injected intrathecally in S and BCP groups, and BAM 8-22 8 nmol/5 μl and MrgC antibody 5 μl were injected intrathecally in BAM and MA groups, respectively, once a day for 7 consecutive days starting from the day 14 after inoculation of the tumor cells.At 1 day before inoculation (T0), before administration (T1) , and at 14, 16 19 and 21 days after inoculation (T2-5, at 0.5 h before the initial administration and 2 h after each administration) , the number of spontaneous flinches (NSF) and mechanical paw withdrawal threshold (MWT) were measured.Five animals selected from each group at each time point were sacrificed, and the lumbar enlargement segments of the spinal cord were removed for determination of MrgC expression in the spinal neurons (by immunofluorescence).Results Compared with group S, NSF was significantly increased, MWT was decreased, and the expression of MrgC was up-regulated at T1-5 in BCP, BAM and MA groups.Compared with group BCP, NSF was significantly decreased, MWT was increased, and the expression of MrgC was up-regulated at T2-5 in group BAM, and NSF was significantly increased, MWT was decreased, and the expression of MrgC was down-regulated at T2-5 in group MA.Conclusion Spinal neuronal MrgC is involved in the maintenance of BCP in mice.

Objective To explore the effect of pre-treatment of subcutaneous injection of ketamine on remifentanil induced hyperalgesia and K+/Cl-cotransporter 2,KCC2) expression on spinal cord of rats.Methods60 male adult SD rats were randomly divided into five groups(n=12 in each group):control group (group C),the incision group(group I),the incision plus remifentanil group(group I+R),the incision plus ketamine group(group I+K) and the incision plus remifentanil and ketamine group(group I+R+K).Mechanical withdrawal threshold (MWT) was evaluated at 24 hours before incision(T0),2 hours,6 hours,24 hours and 48 hours after incision(T1~T4).The lumbar spinal cords of rats were taken out at T4 time point and the KCC2 detected was detected by immunofluorescence analysis and western blot analysis.ResultsCompared with group C(T1(14.5±1.7)g,T2(14.2±1.1)g,T3(13.9±1.8)g,T4(14.2±1.1)g),MWT of other groups at T1 (I(5.6±0.8)g,I+R(3.2±1.0)g,I+K(6.8±1.7)g,I+R+K(5.1±1.6)g),T2 (I(6.9±1.0)g,I+R(4.3±1.2)g,I+K(8.0±1.4)g,I+R+K(6.2±1.5)g),T3 (I(7.6±0.9)g,I+R(5.4±1.1)g,I+K(10.3±1.2)g,I+R+K(7.1±1.1)g),T4 (I(8.9±1.1)g,I+R(7.5±1.4)g,I+K(11.3±1.2)g,I+R+K(8.3±1.2)g)and the expression of KCC2 at T4 decreased (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+R (T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g) decreased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 decreased significantly (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+K (T1(6.8±1.7)g,T2(8.0±1.4)g,T3(10.3±1.2)g,T4(11.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).Compared with group I+R(T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g),MWT of group I+R+K (T1(5.1±1.6)g,T2(6.2±1.5)g,T3(7.1±1.1)g,T4(8.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).ConclusionPre-treatment of subcutaneous injection of ketamine can reduce the hyperalgesia of rats induced by remifentanil and reduce the inhibition of KCC2 expression on dorsal horn of spinal cord.

Objective To evaluate the effect of verapamil on the expression of K+-Cl-cotransporter 2 (KCC2) in spinal dorsal horns during remifentanil-induced hyperalgesia in a rat model of incisional pain.Methods Thirty-two pathogen-free healthy adult male Sprague-Dawley rats,aged 6-7 weeks,weighing 250-300 g,were divided into 4 groups (n=8 each) using a random number table:control group (group C),incisional pain group (group Ⅰ),incisional pain plus remifentanil plus verapamil group (group I+R+ V) and incisional pain plus remifentanil group (group I+R).Normal saline was subcutaneously infused in group C.A 1 cm long incision was made in the plantar surface of the right hindpaw in anesthetized rats in group Ⅰ.Verapamil 5 mg/kg was intraperitoneally injected at 10 min before establishment of the incisional pain model in group I+R+V.In I+R and I+R+V groups,the model of incisional pain was established,and remifentanil was subcutaneously infused for 30 min at a rate of 80 μg · kg-1 · h-1 simultaneously.The mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured at 1 day before establishment of the model (T0) and 2,6,24 and 48 h after establishment of the model (T1-4).The rats were sacrificed after measurement of MWT at T4,and the lumbar enlargement segments of the spinal cord were harvested for determination of the expression of KCC2 by immunofluorescence.Results Compared with group C,the MWT was significantly decreased at T1-4,and the expression of KCC2 was down-regulated in the other groups (P＜0.05).Compared with group Ⅰ,the MWT was significantly decreased at T1-4,and the expression of KCC2 was down-regulated in group I+R (P＜0.05).Compared with group I+R,the MWT was significantly increased at T1-4,and the expression of KCC2 was up-regulated in group I+R+V (P＜0.05).Conclusion The mechanism by which verapamil reduces remifentanil-induced hyperalgesia is related to up-regulation of the expression of KCC2 in spinal dorsal horns in a rat mnodel of incisional pain.

Objective To investigate the expression of insulin-like growth factors-1(IGF-1)in ser-um and phosphorylated IGF-1 receptor in spinal cord in mouse model of bone cancer pain. Methods Sixty male C3H/HeJ mice weighed 18-22 g were randomly divided into Sham group(n=30)and Tumor group(n=30). The mice in Tumor group were inoculated with NCTC fibrosarcoma cells in the right femur bone marrow cavity. Paw withdrawl mechanical threshold(PWMT)and the number of spontaneous flinches(NSF)were measured on 1d before inoculation and on 4 d,7 d,10 d,14 d,21 d after inoculation(n=8). At each time point,the mice of each group were taken blood by removal eyeball and the samples of blood were obtained to detect the expression of IGF-1 by enzyme-linked immunosorbent assay(n=4). The mice after taken blood on 14 d after inoculation were perfused and the samples of spinal cord lumber(L3~5)segment were obtained to detect the expression of phosphorylated IGF-1 receptor by immunofluorescence assay(n=6). Results Com-pared with Sham group,PWMT was significantly decreased(P<0.05)and NSF was significantly increased(P<0.05)on 7~21 d after inoculation. Compared with baseline value and Sham group(baseline value(27.33± 0.52)pg/ml,Sham group(29.11±1.86)pg/ml,(24.51±3.61)pg/ml,(23.33±4.59)pg/ml,(25.29±2.99) pg/ml),the expression of IGF-1 in serum was significantly increased on 7~21 d after inoculation in Tumor group((39.76±3.92)pg/ml,(36.93±2.18)pg/ml,(38.85±2.40)pg/ml,(39.70±2.62)pg/ml). The mean fluorescence intensity of phosphorylated IGF-1 receptor was significantly higher on 14d after inoculation in Tumor group(2.40±0.11)compared with Sham group(0.05±0.01). Conclusion Expression of IGF-1 in serum and phosphorylated IGF-1 receptor in spinal cord were significantly increased in mice with bone cancer pain,and this change may be involved in the development and maintenance of bone cancer pain.

Objective:To compare the clinical efficacy and safety of continuous dosing or alternate-day dosing of apatinib combined with SOX regimen as first-line treatment for patients with advanced gastric cancer.Methods:A total of 52 patients with human epidermal growth factor receptor 2 (HER2) negative and inoperable locally advanced or advanced gastric cancer who were pathologically diagnosed from January 2018 to January 2021 in the Second Affiliated Hospital of Shandong First Medical University were collected. The patients were divided into continuous dosing group and alternate-day dosing group by random number table method. The continuous dosing group received apatinib (250 mg, once a day) combined with SOX regimen (S-1+oxaliplatin); the alternate-day dosing group received apatinib (250 mg, once every other day) combined with SOX regimen. Twenty-one days were a cycle, and the efficacy was evaluated after 2 cycles. After 4-6 cycles, patients with stable disease received apatinib and S-1 for maintenance therapy. The therapeutic effects and adverse reactions of the two groups were compared.Results:The curative effect could be evaluated in 51 patients, including 26 in the continuous dosing group and 25 in the alternate-day dosing group. The disease control rates in the continuous dosing group and the alternate-day dosing group were 84.6% (22/26) and 76.0% (19/25) ( χ2 = 0.60, P = 0.499), and the median progression-free survival time was 7.50 months (95% CI 6.17-8.83 months) and 8.30 months (95% CI 6.99-9.61 months) ( χ2 = 0.71, P = 0.401), and the median overall survival time was 15.50 months (95% CI 11.30-19.69 months) and 15.60 months (95% CI 13.63-17.57 months) ( χ2 = 1.82, P = 0.177). The main adverse reactions in the two groups were leukopenia, thrombocytopenia, hypertension, nausea, vomiting, fatigue, hand-foot syndrome, proteinuria, liver and kidney damage. The incidence rates of ≥grade 3 adverse reactions in the continuous dosing group and the alternate-day dosing group were 42.3% (11/26) and 12.0% (3/25), and the difference was statistically significant ( χ2 = 4.46, P = 0.035). Conclusions:The efficacy of continuous dosing or alternate-day dosing of apatinib combined with SOX regimen as first-line treatment for advanced gastric cancer is similar, but the incidence of ≥grade 3 adverse reactions in alternate-day dosing group is lower, which improves the compliance and tolerance of patients.

Objective To determine risk factors of postoperative pulmonary complications within 1 month in patients undergoing thoracic surgery in Day Care Unit.Methods The total of 200 patients routinely scheduled for VATS under centralized management were enrolled in this study.On the postoperative day 1,lung ultrasound(LUS)was conducted by one physician in the ward.The patients received at least once Chest X-ray or CT in outpatient department within 30 days after discharge.The composite of out-of-hospital PPCs,and the value of LUSS in predicting the PPCs was appraised.Furthermore,we identified the perioperative risk factors associated with PPCs in VATS patients.Results Of 200 recruited VATS patients eligible in the Nanjing Drum Tower Hospital,188 participants received LUS examination and finally completed the 30 days follow-up.Of whom,68 patients developed the varied types of PPCs.Multivariable Logistic regression analysis indicated that comorbidity of immune system disease(P = 0.021),lobar resection(P = 0.031)and the postoperative 24 hours LUSS(P = 0.002)were independent risk factors for PPCs within 30 days after VATS.Conclusion Comorbidity of immune system disease,lobar resection and the postoperative 24 h LUSS were independent risk factors for PPCs within 30 days after VATS.

Specific pathogen-free(SPF)chickens are widely used in the research of avian diseases and vaccines.Vertically transmissible diseases are transmitted to chickens through vertical transmission,seriously affecting their survival rate,increasing production costs,and causing significant economic losses to the poultry industry,while severely impacting the breeding and use of SPF chickens.Therefore,it is crucial for researchers and managers to enhance their understanding of vertically transmissible pathogens in chickens and to develop effective monitoring measures.Quality monitoring is an important part of ensuring the quality of SPF chickens,with pathogen detection being the primary step.Based on this,it is necessary to cultivate qualified SPF chickens through purification methods and biosecurity measures.This paper reviews the major vertically transmissible pathogens in chickens,including viral pathogens,bacterial pathogens and mycoplasmas,as well as their detection methods.This study compares the differences in microbiological testing items and methods for SPF chickens between the U.S.corporate standard and the Chinese national standard.Analysis of the results shows that in both standards,vertically transmissible pathogens such as Escherichia coli,Proteus mirabilis,Salmonella,and avian leukosis are not included in the microbiological testing items for SPF chickens.Instead,these pathogens are characterized by mixed infections,and outbreaks can seriously affect flock health.To produce higher-quality SPF chickens,it is necessary to include these pathogens in the mandatory testing items.The aim of this paper is to help readers understand the relevant standards for microbiological monitoring of SPF chickens,the hazards of vertically transmissible pathogens,and prevention and control strategies,so as to provide a reference for the detection and purification of pathogens in SPF chickens.