Objective To observe the impact of ultrasonic image quality on the consistency of artificial intelligence(Al)assisted diagnosis system and manual measurements of biological indicators of developmental dysplasia of hip(DDH).Methods Hip ultrasonic data of 75 DDH and 345 non-DDH children were retrospectively analyzed,and the quality of ultrasonic images were subjectively scored.An evaluation model of ultrasonic image quality was constructed based on 140 ultrasonic images acquired from 140 cases(group A,containing 25 DDH and 115 non-DDH)using entropy weighting method,the weight of anatomic structures and impact factors related to DDH were obtained.The comprehensive image quality scores of other ultrasonic images acquired from 280 cases(group B,including 50 DDH and 230 non-DDH)were calculated,and the images in group B were classified into grade A,B and C in descending order.The consistency of AI and manual measurements of DDH biological indicators in group B was assessed.Results The weight of each anatomic structure and impact factors of DDH obtained with the model were as follows:The lower edge of iliac branch＞ilium＞glenoid labrum＞bony margin＞femoral head＞motion artifacts.In group B,grade A was observed in 67(9 DDH and 58 non-DDH),grade B was found in 160(26 DDH and 134 non-DDH),while grade C was noticed in 53(15 DDH and 38 non DDH)images.Except for β,femoral head coverage(FHC)and femoral head length diameter,the consistencies between AI and manual measurements of other indicators of DDH were grade A＞B＞C.In group B,AI and manual measurements were more consistent in DDH than in non-DDH cases.Conclusion Ultrasonic image quality affected the consistency between AI and manual measurements of biological indicators of DDH.When image quality was not good enough,further attention should be paid to measurement of FHC and sizes of femoral head.

Objective:To observe the development process of the neuronal synapse in cerebral cortex and basal ganglionic eminence(GE)regions of the mice,and to clarify the differences in the development of excitatory and inhibitory synapses in different brain regions in vivo and in vitro.Methods:The female C57BL/6 mice were euthanized by cervical dislocation from the 13.5th day to the 15.5th day during the pregnancy,and the embryos were collected under the sterile conditions.The cortex and GE regions of brain tissue of the embryonic mice were gradually isolated under microscope.The primary neurons from the embryonic mice were cultured in vitro,and the cell samples were collected on the 3rd,7th,14th,and 21th days,respectively,and regarded as culture 3 d,7 d,14 d,and 21 d groups.The expression levels of postsynaptic density 95(PSD95)and Gephyrin mRNA in the primary neurons from the cortex and GE regions of the mice in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.Immunofluorescence method was used to detect the expression levels of vesicular glutamate transporter 1(vGLUT1),PSD95,vesicular GABA transporter(vGAT),and Gephyrin proteins in the neurons from the cortex and GE regions of the mice in various groups.Immunofluorescence method was also used to detect the expression levels of vGLUT1 and vGAT proteins in the neurons from the cortical and GE regions in brain tissue of the embryonic mice.Results:Compared with culture 3 d group,the expression levels of PSD95 and Gephyrin mRNA in cortex and GE regions of the mice in culture 14 d and 21 d groups were significantly increased(P<0.01).Compared with cortex area,the expression level of Gephyrin mRNA in the neurons from GE region of the mice in culture 14 d group was significantly decreased(P<0.01).The microscope observation results showed that the excitatory and inhibitory synapses in the neurons from cortex and GE regions of the mice in culture 14 d group showed preliminary development,with positive expression of relevant proteins;among them,the excitatory synaptic proteins showed more distinct positive expression in the cortex neurons,and the presynaptic vGLUT1 and postsynaptic PSD95 molecules exhibited co-localization in the cell bodies and protrusions of the cortical neurons;the inhibitory presynaptic vGAT protein and postsynaptic Gephyrin protein in the neurons from GE region also exhibited co-localization in the cell bodies and protrusions,and there were more distinct expressions of the presynaptic molecule proteins than postsynaptic molecule proteins.Compared with cortex region,the levels of vGLUT1 and PSD95 proteins in the neurons from GE region of the mice in culture 14 d group were significantly decreased(P<0.01),while the levels of vGAT and gephyrin proteins were significantly increased(P<0.01).In culture 21 d group,the positive expressions of synaptic protein in the neurons from cortex and GE regions were increased,and the excitatory and inhibitory synapses further matured and enhanced.In the neurons from cortex and GE regions,rich patterns of corresponding pre-and postsynaptic expression were formed in the cell bodies and protrusions,and synapse structures showed gradual,positive development,with more apparent expression of presynaptic molecules compared wih postsynaptic proteins.Compared with cortex region,the levels of vGLUT1 and PSD95 proteins in the neurons from GE region of the mice in culture 21 d group were significantly decreased(P<0.01),and the levels of vGAT and Gephyrin proteins were significantly increased(P<0.01).Compared with cortex region,the expression level of vGLUT1 protein in the neurons from GE region in brain tissue of the embryonic mice was significantly decreased(P<0.01),while the expression level of vGAT protein was significantly increased(P<0.05).Conclusion:There are distinct differences in synaptic development between the neurons from cortex and GE regions,the excitatory synapses develope earlier in the cortical region and the inhibitory synapses develope earlier in the GE region.The region-specific development of synapses suggests that different types of neural diseases with different cell types might originate from different developmental processes.

Objective:To investigate the clinicopathological and genetic characteristics of neuromuscular choristoma-associated desmoid type fibromatosis (NMC-DF).Methods:The clinical morphological and immunohistochemical features of 7 NMC-DF cases diagnosed from January 2013 to January 2023 in Beijing Jishuitan Hospital were retrospectively analyzed. A series of neuromuscular choristoma and neuromuscular choristoma-associated desmoid type fibromatosis were evaluated for CTNNB1 mutations, and hotspot mutations for CTNNB1 were tested in 4 NMC-DF cases using Sanger sequencing.Results:The tumors were collected from 3 females and 4 males, aged 1 to 22 years (mean 7.1 years), involving the sciatic nerve ( n=4), brachial plexus ( n=2) or multiple nerves ( n=1). The course of the disease spanned from 3 months to 10 years. Two cases were recurrent tumors. All the 7 NMC cases showed endoneurial intercalation of mature skeletal muscle fibers among the peripheral nerve fascicles, and the histologic features of the NMC-DF were strikingly similar to the conventional desmoid-type fibromatosis. By immunohistochemistry, all NMC and NMC-DF cases showed aberrant nuclear staining of β-catenin (7/7), the muscle cells in NMC were intensely immunoreactive for desmin, and the admixed nerve fibers were highlighted by NF and S-100 (7/7). Four NMC and NMC-DF had CTNNB1 mutations, 3 c.121A>G (p.T41A) and 1 c.134C>T (p.S45F). Follow-up of the 7 cases, ranging from 22 to 78 months, showed tumor recurrence in 2 patients at 3 and 8 months respectively after the first surgical resection, of which 1 patient underwent above-knee amputation. No recurrence occurred in other cases with tumor excision and neurological reconstruction surgery. There was no metastasis occurred in the 7 cases. Conclusions:NMC is a rare congenital lesion with differentiated mature skeletal muscle tissue found in peripheral nerve fascicles, and approximately 80% of patients with NMC develop a soft tissue fibromatosis. CTNNB1 mutation in the Wnt signaling pathway may be involved in the pathogenesis of NMC and NMC-DF, and S45F mutations seems to have a higher risk of disease progression.

Cancer cachexia is a multi-organ syndrome and closely related to changes in signal communication between organs, which is mediated by cancer cachexia factors. Cancer cachexia factors, being the general name of inflammatory factors, circulating proteins, metabolites, and microRNA secreted by tumor or host cells, play a role in secretory or other organs and mediate complex signal communication between organs during cancer cachexia. Cancer cachexia factors are also a potential target for the diagnosis and treatment. The pathogenesis of cachexia is unclear and no clear effective treatment is available. Thus, the treatment of cancer cachexia from the perspective of the tumor ecosystem rather than from the perspective of a single molecule and a single organ is urgently needed. From the point of signal communication between organs mediated by cancer cachexia factors, finding a deeper understanding of the pathogenesis, diagnosis, and treatment of cancer cachexia is of great significance to improve the level of diagnosis and treatment. This review begins with cancer cachexia factors released during the interaction between tumor and host cells, and provides a comprehensive summary of the pathogenesis, diagnosis, and treatment for cancer cachexia, along with a particular sight on multi-organ signal communication mediated by cancer cachexia factors. This summary aims to deepen medical community’s understanding of cancer cachexia and may conduce to the discovery of new diagnostic and therapeutic targets for cancer cachexia.

Parvalbumin interneurons belong to the major types of GABAergic interneurons. Although the distribution and pathological alterations of parvalbumin interneuron somata have been widely studied, the distribution and vulnerability of the neurites and fibers extending from parvalbumin interneurons have not been detailly interrogated. Through the Cre recombinase-reporter system, we visualized parvalbumin-positive fibers and thoroughly investigated their spatial distribution in the mouse brain. We found that parvalbumin fibers are widely distributed in the brain with specific morphological characteristics in different regions, among which the cortex and thalamus exhibited the most intense parvalbumin signals. In regions such as the striatum and optic tract, even long-range thick parvalbumin projections were detected. Furthermore, in mouse models of temporal lobe epilepsy and Parkinson's disease, parvalbumin fibers suffered both massive and subtle morphological alterations. Our study provides an overview of parvalbumin fibers in the brain and emphasizes the potential pathological implications of parvalbumin fiber alterations.
Mice ; Animals ; Epilepsy, Temporal Lobe/pathology* ; Parvalbumins/metabolism* ; Parkinson Disease/pathology* ; Neurons/metabolism* ; Interneurons/physiology* ; Disease Models, Animal ; Brain/pathology*

Objective: To isolate a bacteriophage against pan-drug resistant Klebsiella pneumoniae in a burn patient, and to study its biological characteristics, genomic information, and effects on bacterial biofilm. Methods: (1) In 2018, pan-drug resistant Klebsiella pneumoniae UA168 (hereinafter referred to as the host bacteria) solution isolated from the blood of a burn patient in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (hereinafter referred to as Ruijin Hospital) was used to isolate and purify the bacteriophage against pan-drug resistant Klebsiella pneumoniae from the sewage of Ruijin Hospital with sewage co-culture method, drip plate method, and double-agar plate method. The bacteriophage was named as phage KP168 and the plaque morphology was observed. (2) The phage KP168 solution was taken for cesium chloride density gradient centrifugation and dialysis, and then the morphology of phage KP168 was observed through transmission electron microscope after phosphotungstic acid negative staining. (3) The phage KP168 solution was taken to determine the lytic ability of the phage KP168 against 20 strains of pan-drug resistant Klebsiella pneumoniae isolated from the burned patients′ blood in Ruijin Hospital by the drip plate method, and then the lysis rate was calculated. (4) The phage KP168 solution at a initial titer of 9.3×10¹¹ plaque-forming unit (PFU)/mL (400 μL per tube) and the host bacteria solution at a concentration of 1×10⁹ colony-forming unit (CFU)/mL (4 mL per tube) were conventionally shaking cultured together for 4 hours at multiplicity of infection (MOI) of 10.000, 1.000, 0.100, 0.010, or 0.001, respectively (1 tube per MOI). The titer of phage KP168 was measured by the double-agar plate method (the measurement method was the same below) to select the optimal MOI. The experiment was repeated three times. (5) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (4 mL per tube) and the phage KP168 solution at an adjusted titer of 5×10⁷ PFU/mL (400 μL per tube) were mixed at the MOI of 0.005. The plaques were counted 0 (immediately), 1, 2, 3, 4, 5, 15, and 30 minutes (1 tube at each time point) after mixing by the double-agar plate method (the counting method was the same below), and the percentage of adsorbed phages was calculated to screen for the optimal adsorption time. The experiment was repeated three times. (6) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (300 μL per tube) and the phage KP168 solution at a titer of 5×10⁸ PFU/mL (60 μL per tube) were mixed at MOI of 0.005 and conventionally shaking cultured after standing for the optimal adsorption time. The phage KP168 titer was measured 0 (immediately), 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 minutes after culture, and a one-step growth curve was drawn. The experiment was repeated three times. (7) The phage KP168 solution at a titer of 2.5×10¹⁰ PFU/mL was left to stand for 1 hour at 37, 40, 50, 60, or 70 ℃ (3 tubes at each time point, 1 mL per tube) for counting the plaques, and then the thermal stability curve was drawn. SM buffer at a pH values of 5.0, 6.0, 7.0, 7.4, 8.0, 9.0, or 10.0 were added to the phage KP168 solution at a titer of 3.0×10¹⁰ PFU/mL, respectively. The mixed solution was left to stand for 1 hour at 37 ℃ (3 tubes of each pH, each tube containing 100 μL phage KP168 solution and 900 μL SM buffer), and then the plaques were counted, and an acid-base stability curve was drawn. (8) The phage KP168 solution was taken for DNA extraction and sequencing after dialysis as in experiment (2). The whole genome was annotated with Prokka to obtain the coding sequence of phage KP168. Nucleotide′s BLAST function was used to proceed nucleic acid sequence alignment for finding a known phage with the highest similarity to the phage KP168 nucleic acid sequence, and Blastx function was used to translate the coding sequence into protein for its function prediction. The comparison with Antibiotic Resistance Genes Database and Virulence Factors Database was proceeded. (9) In a 96-well plate, at a MOI of 1.000, 0.100, 0.010 or 0.001 (3 wells per MOI), 20 μL phage KP168 solution at a initial titer of 5.8×10¹⁰ PFU/mL was added to 200 μL host bacteria solution at a concentration of 1.5×10⁸ CFU/mL (the same concentration below) for co-cultivation for 48 hours. After 200 μL host bacteria solution was left to stand for 48 hours, 20 μL phage KP168 solution at a titer of 1×10^6, 1×10^7, 1×10^8, 1×10^9, or 1×10¹⁰ PFU/mL (3 wells per titer) was added respectively for action for 4 hours. In both experiments, 200 μL host bacteria solution added with 20 μL SM buffer (3 wells) acted as a negative control, and 220 μL LB culture medium (3 wells) acted as a blank control. Absorbance values were measured by a microplate reader, and inhibition/destruction rates of biofilm were calculated. The experiments were both repeated three times. Results: (1) The plaques of phage KP168 successfully isolated and purified were transparent and round, and its diameter was approximately 1.5 mm. (2) The phage KP168 has a regular polyhedron structure with a diameter of about 50 nm and without a tail. (3) The phage KP168 could lyse 13 of 20 strains of Klebsiella pneumoniae from burned patients, with a lysis rate of 65.0%. (4) When MOI was 1.000, the titer was the highest after co-culturing the phage KP168 with the host bacteria for 4 hours, which was the optimal MOI. (5) After the mixing of the phage KP168 with the host bacteria for 4 minutes, the percentage of the adsorbed phage reached the highest, which was the optimal adsorption time. (6) The one-step growth curve showed that during the lysis of the host bacteria by phage KP168, the incubation period was about 10 minutes, and the lysis period was about 40 minutes. (7) With the condition of 40 ℃ or pH 7.4, the number of plaques and the activity of phage KP168 reached the highest. (8) The genome of phage KP168 was a linear double-stranded DNA with a length of 40 114 bp. There were 48 possible coding sequences. It had the highest similarity to Klebsiella phage_vB_Kp1. The most similar known proteins corresponding to the translated proteins of coding sequences contained 23 hypothetical proteins and 25 proteins with known functions. No resistance genes or virulence factor genes were found. The GeneBank accession number was KT367885. (9) After 48 hours of co-cultivation of the phage KP168 and the host bacteria at each MOI, the inhibition rates of biofilm were similar, with an average of about 45%. After the phage KP168 with a titer of 1×10⁹ PFU/mL acted on the biofilm formed by the host bacteria for 4 h, the destruction rate of biofilm was the highest, reaching an average of 42%. Conclusions In this study, a bacteriophage against pan-drug resistant Klebsiella pneumoniae from a burn patient, phage KP168, is isolated from sewage, which belongs to the tailless phage. It has a wide host spectrum, short adsorption time, and short incubation period, with certain thermal and acid-base stability. Its genomic information is clear, and it does not contain resistance genes or virulence factor genes. It also has an inhibitory effect on the formation of bacterial biofilm and a destructive effect on the formed bacterial biofilm.

Objective: To study the effect and mechanism of the extract from grub on the CD40 of microglia cells in rabbit model of central retinal vein occlusion. Methods: The 60 color rabbits were randomly divided into the blank group, model group, Xueshuantong group and sputum group. The Xueshuantong group was intragastrically administered with Xueshuantong tablets suspension 5 mg/ml, the sputum group was given gavage extract 1 g/ml, and the blank group and the model group were intragastrically administered with normal saline once daily. Except for the blank group, the other groups of rabbits were modeled by argon laser irradiation of the retinal vein trunk, and the fundus photography and FFA examination were performed immediately after modeling and at 1, 14 and 28 days after modeling, respectively, at 1, 3, 7, 14 and 28 day, the expression of CD40 in the optic nerve of rabbits was observed by immunohistochemical staining. Results: The FFA results showed that the veins were not filled at 1 day after model establishment, and some veins were not developed at 14 days, and there was no evidence of revascularization at 28 days. Part of the venous in the thrombus group and the sputum group was not filled. The venous filling time was significantly shorter than that in the model group at 14 days, and the venous filling time returned to the normal level at 28 days. The results of immunohistochemistry showed that the number of microglia increased significantly at 1 d in model group. The number of microglia reached the highest peak at 3 d, and the number of microglia decreased at 7 d, 14 d and 28 d. Compared to the model group, at the 3, 7, 14, 28 d, the integrated optical density of CD40 (3 d: 8 908.91 ± 96.30, 6 099.92 ± 273.44 vs. 10 436.4 ± 1 306.8; 7 d: 5 982.06 ± 483.37, 2 957.36 ± 424.19 vs. 8 798.12 ± 444.39; 14 d: 3 225.36 ± 468.88, 342.04 ± 64.56 vs. 5 356.74 ± 439.16; 28 d: 756.97 ± 80.17, 72.85 ± 11.06 vs. 4 215.27 ± 361.00) in the Xueshuantong group and sputum group significantly decreased (P<0.05). Conclusions The thrombus and sputum extract can inhibit the activation of microglia to varying degrees, and the sputum extract is moreinhibitory effect.

Objective To find out the clinical indicators related to prognosis in patients with acute mushroom poisoning, and approach its correlation with prognosis. Methods Clinical data of patients with mushroom poisoning admitted to the First Hospital of China Medical University, the Ninth People's Hospital of Shenyang, Xiuyan Central People's Hospital, and Fushun Central Hospital from August 2015 to August 2017 were retrospectively analyzed. The biochemical indicators within 24 hours after admission, sequential organ failure assessment (SOFA) score, model for end-stage liver disease (MELD) score, whether plasmapheresis (PE) was carried out or not and 28-day prognosis of patients were collected. According to prognosis, the patients were divided into death group and survival group, and the differences in above parameters between the two groups were compared. Spearman or Pearson correlation method was used to analyze the relationship between MELD score and prognosis. Receiver operating characteristic (ROC) curve was used to analyze the prognostic value of MELD score for prognosis. Further analysis of the patients receiving PE treatment was conducted. Results A total of four Liaoning hospitals with 89 patients with mushroom poisoning were enrolled, with 6 died within 28 days, and 83 survived. There were 17 patients with severely impaired liver and coagulant functions accepted PE treatment, with 6 patients died within 28 days, and 11 survived. ① In 89 patients, compared with survival group, MELD score, prothrombin time (PT), activated partial thromboplastin time (APTT), total bilirubin (TBil), international normalized ratio (INR), blood glucose (Glu), alanine aminotransferase (ALT), γ-glutamyltransferase (GGT) in death group were significantly increased [MELD score: 32.34 (28.31, 41.06) vs. 8.76 (3.77, 21.19), PT (s): 53.5 (52.4, 113.2) vs. 14.5 (13.8, 19.5), APTT (s): 58.6 (48.9, 70.8) vs. 36.9 (34.4, 43.2), TBil (μmol/L): 134.8 (31.3, 155.6) vs. 21.5 (15.1, 41.4), INR: 6.0 (5.6, 14.7) vs. 1.2 (1.1, 1.5), Glu (mmol/L): 9.2 (9.0, 11.0) vs. 6.6 (5.7, 7.8), ALT (U/L):5 923.0 (1 105.0, 6 000.0) vs. 35.0 (18.0, 1 767.0), GGT (U/L): 49.0 (32.0, 57.0) vs. 25.0 (16.0, 41.0), all P < 0.05], but the prothrombin activity (PTA), albumin (ALB), serum Na+, Cl- were significantly decreased [PTA: 13.0% (6.0%, 14.0%) vs. 80.0% (61.0%, 87.0%), ALB (g/L): 31.1 (29.8, 39.0) vs. 42.4 (37.9, 44.3), Na+(mmol/L): 126.5 (122.4, 131.0) vs. 137.0 (134.9, 141.0), Cl- (mmol/L): 93.5 (87.6, 95.0) vs. 104.0 (101.3, 106.0), all P < 0.05]. Spearson correlation analysis showed that MELD score of patients with mushroom poisoning was positively correlated with the 28-day mortality (r = 0.423, P = 0.001). ROC curve analysis showed that the area under ROC curve (AUC) of MELD score for prognosis of patients with mushroom poisoning was 0.926; when the cut-off value was 27.30, the sensitivity was 100%, and the specificity was 84.3%. ② In 17 patients who accepted PE treatment, compared with survival group, the MELD score, TBil, Glu, and ALT in the death group were significantly increased [MELD score: 36.81±5.18 vs. 29.01±5.23, TBil (μmol/L): 145.2±13.9 vs. 93.2±44.0, Glu (mmol/L): 9.1±1.9 vs. 6.0±2.7, ALT (U/L): 5 961.5±44.5 vs. 3 932.9±1 625.7, all P < 0.05], and Cl- was significantly lowered (mmol/L: 94.3±1.2 vs. 100.5±5.7, P < 0.05), but SOFA score showed no significant difference (5.83±2.71 vs. 5.91±1.58, P > 0.05). Correlation analysis showed that the MELD score in patients with mushroom poisoning who accepted PE treatment was positively correlated with 28-day mortality (r = 0.355, P = 0.001), but no correlation with SOFA score was found (r = 0.427, P = 0.087). ROC curve analysis showed that the AUC of MELD score in the prediction of mushroom poisoning patients undergoing PE treatment was 0.545; when the cut-off value was 32.19, the sensitivity was 33.3%, and the specificity was 100%. Conclusions In mushroom poisoning patients, especially those undergoing PE treatment, the higher the MELD score, the higher the mortality is. MELD score could assess the prognosis of patients with acute mushroom poisoning.

Objective To evaluate the safety and feasibility of INTRABEAM in breast conserving surgery for early stage breast cancer.Methods Clinical data of 43 cases of early breast cancer undergoing INTRA-BEAM intraopetative radiotherapy from Jan.2013 to Dec.2014 were retrospectively analyzed.All cases underwent breast conserving surgery combined with 20 Gy INTRABEAM intraoperative radiotherapy.The postoperative incision,incidence of local complications and acute radiation injury were recorded after surgery.Breast recovery,the cosmetic effects,early overall survival,recurrence-free survival,and non-metastatic survival were followed up.Results All cases were given breast conserving surgery associated with INTRABEAM intraoperative radiotherapy (20 Gy),with median radiotherapy time of 31 mins ranging from 25 to 39 mins.Five cases underwent postoperative whole breast irradiation.Major early complications included incision infection (1 case),postoperative effusion (5 cases),radiation area skin pain (4 cases).The short-term follow-up survey showed that the satisfaction rate was 93.0％.The overall survival rate,recurrence free survival rate and metastasis free survival rate was 100％ respec tively.Conclusion Breast conseving surgery combined with INTRABEAM intraoperative radiotherapy for early breast cancer patients is safe and feasible.