Objective:To explore the consistency of claudin 18.2 immunohistochemistry (IHC) using 4 different clone antibodies in gastric adenocarcinoma.Methods:A total of 226 gastric adenocarcinomas diagnosed and treated at the Drum Tower Hospital Affiliated to Nanjing University Medical College between January 2022 to March 2023 were included in this study. The cohort consisted of 165 males and 61 females, with a mean age of (61.3±12.1) years. Tumor tissues from radical resection specimens were collected for tissue microarrays. IHC detection of claudin 18.2 was performed using the EnVision method, utilizing 4 clones of antibody: OTIR157B5, 43-14A, EPR19202 and D313D22. The results were interpreted based on both the intensity of staining on tumor cell membranes and the percentage of positive tumor cells relative to the total tumor cells.Results:The positive cutoff value was set as moderately to strongly linear membrane staining in ≥75% of all viable invasive tumor cells, and clone OTIR157B5 demonstrated the highest positive expression rate at 52.2% (118/226). Additionally, the clones OTIR157B5, 43-14A, and EPR19202 were consistently and strongly positive, with all agreement rates of Cohen κ exceeding 0.8. In gastric adenocarcinoma and its three Lauren subtypes, OTIR157B5 exhibited clear membranous localization.Conclusions:Clone OTIR157B5 of claudin 18.2 antibody shows the highest rate of moderately to strongly linear membrane-positive staining, accounting for ≥75% of all viable invasive tumor cells, and clones 43-14A and EPR19202 show strong consistency and high sensitivity.

Objective:To investigate the neural regulatory mechanism of parvalbumin-positive interneurons(PV-INs)in the dentate gyrus(DG)involved in the impairment of spatial recognition memory by exercise-induced chronic fatigue.Methods:Male Sprague-Dawley(SD)rats were divided into control group and fatigue group by random num-ber method.A three-level incremental load treadmill training program was selected to establish a chronic exhaustion exercise fatigue model.The spatial recognition memory ability of rats was tested by novel object recognition test.The ac-tivation levels and quantitative changes of astrocytes(AS)and PV-INs in the DG region was observed and quantified through immunofluorescence staining and immunohistochemical staining.The phosphorylation level of calcium/calmodu-lin-dependent protein kinase Ⅱ(CaMK Ⅱ)in the hippocampus was detected by Western blot.Results:In the test of novel object recognition,the exploration time of novel object was reduced in the fatigue group,and the discrimination index was significantly lower than that in the control group(P＜0.01).Immunohistochemical staining showed that PV-INs in the DG region of fatigue rats were lighter and fewer than those in the control group,the fibers were short and sparse,and the positive cell density and average optical density of cells were significantly lower than those in the control group(P＜0.01).Immunofluorescence staining showed that AS was significantly activated,glial fibrillary acidic pro-tein(GFAP)was stained deeply,and the cell processes were dense and elongated in the DG region of fatigue rats.The positive cell density and mean fluorescence intensity were significantly higher than those of the control group(P＜0.01).The results of Western blot showed that the phosphorylation level of CaMK Ⅱ protein in the hippocampus of the fatigue group was significantly reduced than that of the control group(P＜0.01).Conclusion:Exercise-induced chro-nic fatigue inhibited PV-INs in the rat hippocampal DG region.The excessive activation of AS following exercise fatigue may be a major contributor to this PV-INs suppression.Concurrently,reduced phosphorylation levels of CaMK Ⅱ protein were observed in hippocampal tissue.These alterations ultimately impaired spatial recognition memory in the rats.

Objective To explore the effect of bionic electrical stimulation(BES)combined with resistance tightening leg training on postpartum pelvic floor dysfunction(PFD).Methods A total of 205 patients with PFD were divided into the BES group(n=102)and the combined group(n=103)by random number table method.All subjects received routine pelvic floor muscle training.In addition,the BES group was treated with BES intervention,and the combined group was treated with BES combined with resistance tightening leg training,with 8 weeks of treatment.The classification of pelvic floor muscle strength,electrophysiological indexes of pelvic floor function,urodynamics and stress urinary incontinence(SUI)classification/pelvic organ prolapse(POP)degree were compared between the two groups before and after treatment.Results After treatment,the pelvic floor muscle strength classification,SUI classification and POP degree in the combined group were significantly better than those in the BES group(P<0.05).After treatment,ClassⅠ/ClassⅡmuscle fiber fatigue,vaginal dynamic pressure and maximal electromyography(EMG)value of fast muscle in the combined group were(-1.20±0.35)%,(-0.90±0.30)%,(76.30±5.51)cmH2 O,(43.00±5.82)μV,respectively,which were all significantly higher than those in the BES group(P<0.05).After treatment,the indexes of urodynamics in both groups were significantly improved,and the combined group was better than the BES group(P<0.05).Conclusions BES combined with resistance tightening leg training can significantly improve pelvic floor muscle strength,muscle fiber fatigue and urodynamics in patients with PFD,thus alleviating SUI/POP symptoms.

Objective:To investigate the neural regulatory mechanism of parvalbumin-positive interneurons(PV-INs)in the dentate gyrus(DG)involved in the impairment of spatial recognition memory by exercise-induced chronic fatigue.Methods:Male Sprague-Dawley(SD)rats were divided into control group and fatigue group by random num-ber method.A three-level incremental load treadmill training program was selected to establish a chronic exhaustion exercise fatigue model.The spatial recognition memory ability of rats was tested by novel object recognition test.The ac-tivation levels and quantitative changes of astrocytes(AS)and PV-INs in the DG region was observed and quantified through immunofluorescence staining and immunohistochemical staining.The phosphorylation level of calcium/calmodu-lin-dependent protein kinase Ⅱ(CaMK Ⅱ)in the hippocampus was detected by Western blot.Results:In the test of novel object recognition,the exploration time of novel object was reduced in the fatigue group,and the discrimination index was significantly lower than that in the control group(P＜0.01).Immunohistochemical staining showed that PV-INs in the DG region of fatigue rats were lighter and fewer than those in the control group,the fibers were short and sparse,and the positive cell density and average optical density of cells were significantly lower than those in the control group(P＜0.01).Immunofluorescence staining showed that AS was significantly activated,glial fibrillary acidic pro-tein(GFAP)was stained deeply,and the cell processes were dense and elongated in the DG region of fatigue rats.The positive cell density and mean fluorescence intensity were significantly higher than those of the control group(P＜0.01).The results of Western blot showed that the phosphorylation level of CaMK Ⅱ protein in the hippocampus of the fatigue group was significantly reduced than that of the control group(P＜0.01).Conclusion:Exercise-induced chro-nic fatigue inhibited PV-INs in the rat hippocampal DG region.The excessive activation of AS following exercise fatigue may be a major contributor to this PV-INs suppression.Concurrently,reduced phosphorylation levels of CaMK Ⅱ protein were observed in hippocampal tissue.These alterations ultimately impaired spatial recognition memory in the rats.

Objective To explore the effect of bionic electrical stimulation(BES)combined with resistance tightening leg training on postpartum pelvic floor dysfunction(PFD).Methods A total of 205 patients with PFD were divided into the BES group(n=102)and the combined group(n=103)by random number table method.All subjects received routine pelvic floor muscle training.In addition,the BES group was treated with BES intervention,and the combined group was treated with BES combined with resistance tightening leg training,with 8 weeks of treatment.The classification of pelvic floor muscle strength,electrophysiological indexes of pelvic floor function,urodynamics and stress urinary incontinence(SUI)classification/pelvic organ prolapse(POP)degree were compared between the two groups before and after treatment.Results After treatment,the pelvic floor muscle strength classification,SUI classification and POP degree in the combined group were significantly better than those in the BES group(P<0.05).After treatment,ClassⅠ/ClassⅡmuscle fiber fatigue,vaginal dynamic pressure and maximal electromyography(EMG)value of fast muscle in the combined group were(-1.20±0.35)%,(-0.90±0.30)%,(76.30±5.51)cmH2 O,(43.00±5.82)μV,respectively,which were all significantly higher than those in the BES group(P<0.05).After treatment,the indexes of urodynamics in both groups were significantly improved,and the combined group was better than the BES group(P<0.05).Conclusions BES combined with resistance tightening leg training can significantly improve pelvic floor muscle strength,muscle fiber fatigue and urodynamics in patients with PFD,thus alleviating SUI/POP symptoms.

Objective:To explore the consistency of claudin 18.2 immunohistochemistry (IHC) using 4 different clone antibodies in gastric adenocarcinoma.Methods:A total of 226 gastric adenocarcinomas diagnosed and treated at the Drum Tower Hospital Affiliated to Nanjing University Medical College between January 2022 to March 2023 were included in this study. The cohort consisted of 165 males and 61 females, with a mean age of (61.3±12.1) years. Tumor tissues from radical resection specimens were collected for tissue microarrays. IHC detection of claudin 18.2 was performed using the EnVision method, utilizing 4 clones of antibody: OTIR157B5, 43-14A, EPR19202 and D313D22. The results were interpreted based on both the intensity of staining on tumor cell membranes and the percentage of positive tumor cells relative to the total tumor cells.Results:The positive cutoff value was set as moderately to strongly linear membrane staining in ≥75% of all viable invasive tumor cells, and clone OTIR157B5 demonstrated the highest positive expression rate at 52.2% (118/226). Additionally, the clones OTIR157B5, 43-14A, and EPR19202 were consistently and strongly positive, with all agreement rates of Cohen κ exceeding 0.8. In gastric adenocarcinoma and its three Lauren subtypes, OTIR157B5 exhibited clear membranous localization.Conclusions:Clone OTIR157B5 of claudin 18.2 antibody shows the highest rate of moderately to strongly linear membrane-positive staining, accounting for ≥75% of all viable invasive tumor cells, and clones 43-14A and EPR19202 show strong consistency and high sensitivity.

Objective To analyze the effects of massage therapy on the inflammatory state and lung function of pediatric refractory Mycoplasma pneumonia (RMPP). Methods A total of 320 children with RMPP treated in Hebei Seventh People′s Hospital from September 2022 to August 2023 were selected in the study. According to different treatment methods, they were divided into two groups, with 160 cases in each group. The western medicine group was given conventional anti-inflammatory and other symptomatic treatment, while the massage group was given dialectical massage treatment based on the western medicine group. The clinical efficacy, duration of disappearance of clinical symptoms, pulmonary function indexes, serum inflammatory factors, immune function indexes and total incidence of adverse reactions were compared between the two groups. Results The total effective rate of massage group was higher than that of control group (P < 0.05). The disappearance time of fever, cough, sputum and lung moist rales in observation group was shorter than that in control group (P < 0.05). After treatment, the peak expiratory flow (PEF), maximum mid-expiratory flow (MMEF) and forced vital capacity (FVC) in the massage group were higher than those in the control group (P < 0.05). The levels of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the massage group were lower than those in the control group after treatment (P < 0.05). After treatment, CD3⁺, CD4⁺, CD4⁺/CD8⁺ in the massage group were higher than those in the control group (P < 0.05), and CD8⁺ was lower than that in the control group (P < 0.05). There was nosignificant difference in the total incidence of adverse reactions between the massage group and the western medicine group (P>0.05). Conclusion Massage based on syndrome differentiation combined with western medicine can effectively relieve fever, cough and other symptoms of RMPP children, reduce inflammatory response, improve lung function and immune function, and has less adverse reactions.

This experiment aims to investigate the effects of a dietary supplement of herbal tea resi-due on white feather broilers in the prospectives of growth performance,serum indexes and intesti-nal immune indexes.A total of 280 1-day-old white feather broilers with similar body mass and good health were randomly divided into 4 groups with 5 replicates per group and 14 birds per repli-cate.Group Ⅰ(control group)was fed a basal diet.Groups Ⅱ,Ⅲ and Ⅳ were fed a diet supple-mented with 2％,4％and 6％herbal tea residue powder,respectively.The feeding lasted for 49 days,and was divided into 2 phases from 1 to 21 days of age and 22 to 49 days of age.Blood,tissue and mucosa of the duodenum and jejunum were collected on 21 d and 49 d.Total cholesterol(T-CHO),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)and the activities of aspartate aminotransferase(AST)and alanine amin-otransferase(ALT)in the serum were examined.The villus height and crypt depth of the jejunum were observed and measured by morphology.The mRNA expression levels of intestinal interleukin-1β(IL-1β),interleukin-2(IL-2),interleukin-4(IL-4),interleukin-10(IL-10)and tumor necrosis factor-α(TNF-α)were determined by qPCR.The results showed that compared with group Ⅰ,the mass of group Ⅲ on 21 d and 49 d significantly increased(P＜0.05).The average daily feed intake of 22-49 d and 1-49 d significantly increased(P＜0.05).The average daily mass gain of 1-21 d,22-49 d and 1-49 d significantly increased(P＜0.05).In the serum of 21 d,compared with group Ⅰ,T-CHO in group Ⅱ significantly decreased(P＜0.05).TG in groups Ⅱ,m and Ⅳ significantly de-creased(P＜0.05).LDL-C and AST activity in group Ⅳ significantly decreased(P＜0.05).In the serum of 49 d,T-CHO in group Ⅲ,LDL-C in group Ⅳ,HDL-C of groups Ⅲ and Ⅳ significantly increased(P＜0.05),and the ALT activity of these two groups decreased(P＜0.05).AST activity in group Ⅱ significantly decreased(P＜0.05).In the duodenum of 21 d,compared with group Ⅰ,the expressions of TNF-α in group Ⅱ was significantly decreased(P＜0.05).In the duodenum of 49 d,the expression of IL-10 in group Ⅱ was significantly increased(P＜0.05),and the expression of IL-2 in groups Ⅲ and Ⅳ was significantly decreased(P＜0.05).In the jejunum of 49 d,the ex-pression of IL-10 in group Ⅱ was increased(P＜0.05),and the expression of IL-2 in groups Ⅲand Ⅳ was significantly decreased(P＜0.05).In conclusion,dietary herbal tea residue can regulate circulating lipid level and enhance intestinal immunity of white feather broilers without affecting their normal growth performance.This experiment also suggested that the a 2％-4％supplementa-tion of herbal tea residues to white feather broilers was of good effect.

The study adopted muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid and electrostimulation to determine its effects on mouse growth and sow production performance.One hundred and fifty four-week-old C57 BL/6 female mice were randomly divided into 6 groups of 5 replicates each.Muscle single-injection followed by electrostimulation was performed.The con-trol group received an empty plasmid injection(80 μg/kg),while the treatment groups received pVAX1-SP-GHRH(1-44)plasmid injections(20,40,80,120,160 μg/kg).Twenty healthy preg-nant sows were randomly divided into 2 groups,each with 10 sows.Electrostimulation treatment was applied to the semimembranosus muscle of the pregnant sows after a single injection.The con-trol group received physiological saline injection,while the plasmid group received a 2 mg pVAX1-SP-GHRH(1-44)expression plasmid injection.Mouse weight,feed intake,and serum GHRH and IGF-1 levels were measured at days 0,7,14,21,and 28 after injection.Pregnant sows were bled via the tail vein at days 0,14,28,and 42 after injection,and their serum was separated to measure serum GHRH and IGF-1 levels.The birth weight,placental weight,number of piglets born,number of healthy piglets,number of weak piglets,number of deformed piglets,number of stillborn piglets,and number of mummified piglets were recorded at day 14.The mouse study re-sults showed that muscle injection of pVAX1-SP-GHRH(1-44)plasmid followed by electrostim-ulation could promote mouse feeding and increase weight gain(P＜0.05),significantly increase mouse serum GHRH and IGF-1 levels(P＜0.05),and maintain its effects until day 21.The results of the pregnant sow study showed that the average birth weight of the piglets in the plasmid group was significantly increased(P＜0.01),and the placenta weight was significantly increased(P＜0.05).The serum GHRH and IGF-1 concentrations in the plasmid group sows were significantly increased(P＜0.01).The study results showed that muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid followed by electrostimulation could promote mouse feeding and increase weight gain,and also significantly improve the average birth weight and placental weight of the piglets in pregnant sows.