BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

ObjectiveTo investigate the efficacy， safety， and cost-effectiveness of endoscopic ultrasound （EUS）-guided coil placement combined with tissue adhesive injection in the treatment of gastric varices with spontaneous shunt. MethodsA retrospective analysis was performed for the patients with acute gastric variceal bleeding and spontaneous portosystemic shunt who were hospitalized and received balloon-occluded retrograde transvenous obliteration （BRTO） combined with endoscopic tissue adhesive injection or EUS-guided coil placement combined with tissue adhesive injection in Xiangyang Central Hospital from March 2019 to September 2022. The two surgical procedures were compared in terms of efficacy （technical success rate， 5-day rebleeding rate， 1-year rebleeding rate， and time to rebleeding）， safety （the incidence rate of ectopic embolism， the amount of tissue adhesive used， and the amount of lauromacrogol used）， and cost-effectiveness （hospital costs and length of hospital stay）. The t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The Kaplan-Meier method was used to estimate the rebleeding. The chi-square test was used for comparison of categorical data between two groups. ResultsA total of 25 patients received successful EUS-guided coil placement and tissue adhesive injection， with a technical success rate of 100%， a median amount of 2.5 mL tissue adhesive used， a median amount of 11.0 mL lauromacrogol used， a mean length of hospital stay of 14.88±3.21 days， a mean hospital cost of 32 660.00±4 602.07 yuan， and a 5-day rebleeding rate of 0%； among these patients， 2 were lost to follow-up， and 23 patients with complete follow-up data had an incidence rate of ectopic embolism of 0% and a median time to rebleeding of 689 days. A total of 14 patients underwent modified BRTO combined with endoscopic tissue adhesive injection， with a technical success rate of 100%； a median amount of 5.0 mL tissue adhesive used during surgery， which was significantly higher than that used in EUS （U=39.000， P<0.001）； a median amount of 10.5 mL lauromacrogol used during surgery； a mean length of hospital stay of 15.38±4.94 days； a mean hospital cost of 57 583.47±18 955.40 yuan， which was significantly higher than that used in EUS （t=-6.310， P<0.001）； a 5-day rebleeding rate of 0%. No patient was lost to follow-up， and all 14 patients had an incidence rate of ectopic embolism of 0% and a median time to rebleeding of 244.50 days， with no significant difference between the two groups （χ²=1.448， P=0.229）. ConclusionEUS-guided coil placement combined with tissue adhesive injection is a relatively safe and effective technique for the treatment of gastric variceal bleeding and has a high technical success rate， a low incidence rate of serious adverse events， and similar efficacy to BRTO， with higher safety and cost-effectiveness.

ObjectiveExosomes are microvesicles which could be secreted by all cell types with diameters between 30 and 150 nm. It was widely distributed in body fluids including blood, urine, and breast milk. Exosomes are considered as potential biomarkers and drug carriers by reason of containing nucleic acids, lipids, proteins and other bioactive molecules. Milk-derived exosomes have been widely used as drug delivery carriers to treat targeted diseases with a lower cost, higher biocompatibility and lower immunogenicity. Until now, there is no research about the milk-derived exosomes phosphorylation to reveal the difference of protein phosphorylation in different species of milk. To investigate the pathways and proteins with specific functions, phosphorylated proteomic analysis of milk-derived exosomes from different species is performed, and provide new ideas for exploring diversified treatments of disease. MethodsWhey and exosomes derived from bovine, porcine and caprine milk were performed for proteomics and phosphoproteomics analysis. The relationship between milk exosome proteins from different species and signaling pathways were analyzed using bioinformatics tools. ResultsA total of 4 191 global proteins, 1 640 phosphoproteins and 4 064 phosphosites were identified from 3 species of milk-derived exosomes, and the exosome proteins and phosphoproteins from different species were significantly higher than those of whey. Meanwhile, some special pathways were enriched like Fcγ-mediated phagocytosis from bovine exosomes, pathways related with neural and immune system from caprine exosomes, positive and negative regulation of multiple activities from porcine exosomes. ConclusionIn this study, the proteomic and phosphoproteomic analyses of exosomes and whey from bovine, porcine and caprine milk were carried out to reveal the difference of composition and related signaling pathways of milk exosome from different species. These results provided powerful support for the application of exosomes from different milk sources in the field of disease treatment.

The compound (E)-1-(4-(3-(5-chloro-6-oxo-3,6-dihydropyridin-1(2H)-yl)-3-oxo-propyl-1-ene-1-yl) phenyl)-3-(4-fluorophenyl) urea (C12), a novel derivative of piperlongumine previously synthesized by our research group, was investigated in this study to examine its effects on human non-small cell lung cancer cell line H1299 in vitro and elucidate its potential mechanism of action. The impact of C12 on the proliferation, migration, and invasion abilities of H1299 cells were assessed using methyl thiazolyl tetrazolium (MTT) assay, wound healing assay, cloning formation assay, and Transwell assay. Flow cytometry was employed to evaluate the influence of C12 on cell cycle progression, reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), and apoptosis induction in H1299 cells. Western blot analysis was conducted to investigate the expression levels of p21, Cyclin B1, CDK1, Bax, Bcl-2, JNK, p-JNK, Erk1/2, p-Erk1/2, p38 and p-p38 proteins for exploring the anti-tumor mechanism underlying C12's actions. The results demonstrated that C12 exerted inhibitory effects on the proliferation, migration, and invasion capacities of H1299 cells in a time-dependent and concentration-dependent manner. Moreover, C12 induced G2/M phase arrest in the cell cycle, reduced MMP levels, elevated ROS production, and triggered apoptotic processes. Flow cytometry analysis revealed that C12 downregulated Cyclin B1 and CDK1 protein expressions, resulting in G2/M phase arrest. C12 also upregulated Bax/Bcl-2 ratio, promoting apoptosis. Furthermore, C12 activated MAPK signaling pathway by enhancing phosphorylation levels of JNK, Erk1/2, and p38 proteins. In conclusion, C12 significantly suppressed proliferation, migration, and invasion capabilities while inducing cell cycle arrest and apoptosis in H1299 cells. These effects may be attributed to activation of the MAPK signaling pathway.

Objective To develop an automatic urine volume detection and collection device to solve the problems of routine urine test.Methods An automatic urine volume detection and collection device was developed with the components of a main control system,a detection system,a prompting system and a grasping and moving system.The main control system consisted of two STM32 microcontrollers and a reset switch;the detection system was made up of a weighing module,an infrared module and indicator lights,which had its urine volume automatic detection algorithm developed based on the Keil5 platform;the prompting system realized voice broadcasting through the voice module fixed on the back panel of the box;the grasping and moving system was composed of a rail drive motor(86CM stepper motor),a photoelectric switch and a motorized gripper.Results The device developed tested urine samples with an accuracy of 99.44％,and could collect qualified samples automatically and quickly.Conclusion The device developed detects urine volume and collects samples automatically,and enhances the accuracy and efficiency of urine examination.[Chinese Medical Equipment Journal,2024,45(4):66-69]

Objective To propose an evaluation and screening indicator for the reliability of emergency ventilators.Methods Firstly,a regression model was used to clean the data and remove noise to ensure the accuracy of regression analysis.Then,four groups of highly correlated data combinations,including inspiratory tidal volume-minute expiratory volume,peak airway pressure-minute expiratory volume,peak airway pressure-inspiratory tidal volume and positive end-expiratory pressure(PEEP)-mean airway pressure,were determined with the methods of curve fitting and transfer function,and the difference between the theoretical output and the actual output of the data combinations was regarded as an indicator to judge whether the ventilator functioned well or not;finally,the indicator proposed was applied to single and multiple ventilators,and the feasibility of the indicator was determined by the proportion of the ventilators functioning well.Results The evaluation results with a single ventilator showed the four groups of data combinations had the actual output fitted well with the theoretical output,and all the differences between the theoretical output and the actual output were lower than the threshold;the results with multiple ventilators indicated there were 71.49％ventilators functioning well,which was very close to the actual result that 72％ventilators behaved well.Conclusion The evaluation and screening indicator for emergency ventilators has high feasibility,and theoretical support is provided for reliability assessment and selection of series of emergency treatment equipment.[Chinese Medical Equipment Journal,2024,45(7):8-16]

Objective To investigate the levels of serum folate and vitamin B12(VB12)and their association with the level of neurodevelopment in preschool children with autism spectrum disorder(ASD).Methods A total of 324 ASD children aged 2-6 years and 318 healthy children aged 2-6 years were recruited.Serum levels of folate and VB12 were measured using chemiluminescent immunoassay.The Social Responsiveness Scale and the Childhood Autism Rating Scale were used to assess the core symptoms of ASD children,and the Gesell Developmental Schedule was employed to evaluate the level of neurodevelopment.Results The levels of serum folate and VB12 in ASD children were significantly lower than those in healthy children(P＜0.05).Serum folate levels in ASD children were positively correlated with gross and fine motor developmental quotients(P＜0.05),and serum VB12 levels were positively correlated with adaptive behavior,fine motor,and language developmental quotients(P＜0.05).In ASD children aged 2 to＜4 years,serum folate levels were positively correlated with developmental quotients in all domains(P＜0.05),and serum VB12 levels were positively correlated with language developmental quotient(P＜0.05).In male ASD children,serum VB12 levels were positively correlated with language and personal-social developmental quotients(P＜0.05).Conclusions Serum folate and VB12 levels in preschool ASD children are lower than those in healthy children and are associated with neurodevelopmental levels,especially in ASD children under 4 years of age.Therefore,maintaining normal serum folate and VB12 levels may be beneficial for the neurodevelopment of ASD children,especially in ASD children under 4 years of age.[Chinese Journal of Contemporary Pediatrics,2024,26(4):371-377]

Objective:To explore the clinical characteristics,molecular characteristics,treatment and prognosis of pediatric Philadelphia chromosome-like acute lymphoblastic leukemia(Ph-like ALL)with a therapeutic target.Methods:A total of 27 patients of Ph-like ALL with targeted drug target were initially diagnosed in Children's Hospital of Soochow University from December 2017 to June 2021.The data of age,gender,white blood cell(WBC)count at initial diagnosis,genetic characteristics,molecular biological changes,chemotherapy regimen,different targeted drugs were given,and minimal residual disease(MRD)on day 19,MRD on day 46,whether hematopoietic stem cell transplantation(HSCT)were retrospective analyed,and the clinical characteristics and treatment effect were summarized.Survival analysis was performed by Kaplan-Meier method.Results:The intensity of chemotherapy was adjusted according to the MRD level during induced remission therapy in 27 patients,10 patients were treated with targeted drugs during treatment,and 3 patients were bridged with HSCT,1 patient died and 2 patients survived.Among the 24 patients who did not receive HSCT,1 patient developed relapse,and achieved complete remission(CR)after treatment with chimeric antigen receptors T cells(CAR-T).The 3-year overall survival,3-year relapse-free survival and 3-year event-free survival rate of 27 patients were(95.5±4.4)％,(95.0±4.9)％and(90.7±6.3)％respectively.Conclusion:Risk stratification chemotherapy based on MRD monitoring can improve the prognosis of Ph-like ALL in children,combined with targeted drugs can achieve complete remission as soon as possible in children whose chemotherapy response is poor,and sequential CAR-T and HSCT can significantly improve the therapeutic effect of Ph-like ALL in children whose MRD is continuously positive during induced remission therapy.

Objective:To explore the effects of hypoxia on traumatic brain swelling (TBS) in rats with acute subdural hematoma (ASDH).Methods:Forty-five SD rats were divided into 5 groups according to the random number table method, with 9 rats in each group: sham surgery normal oxygen group which underwent sham surgical procedures and were placed in a closed container with ventilation, sham surgery hypoxia group which underwent sham surgical procedures and were placed in a closed container with oxygen volume fraction of 8% for hypoxia induction, ASDH normal oxygen group which made into the ASDH model and placed in a closed container with ventilation, ASDH hypoxia group were made into the ASDH models and placed in a closed container with oxygen volume fraction of 8% for hypoxia induction, and ASDH hypoxia+oxygen inhalation group which inhaled oxygen continuously with oxygen volume fraction of 40% after being made into the ASDH models and induced for hypoxia. Six rats were selected from each group immediately after the modeling and craniotomy was performed to observe the brain swelling during the surgery and evaluate the degree of TBS. Microvascular blood flow was observed by laser speckle imaging system before modeling, before craniotomy, and immediately after craniotomy. The remaining 3 rats in each group were killed directly after modeling and brain tissue specimens were collected. The expression levels of pericellular protein α-smooth muscle actin (α-SMA) and platelet-derived growth factor receptor-β (PDGFR-β) at 0, 30 and 60 minutes after modeling were detected through Western blot analysis. The expression levels of α-SMA, PDGFR-β and microvascular marker platelet-endothelial cell adhesion molecule 31 (CD31) at 0 minute after modeling were tested through immunofluorescent staining.Results:No brain bulge was observed in the sham surgery normal oxygen group. The height of brain bulge in sham surgery hypoxia group was 0.5(0.0, 1.0)mm, with no significant difference from that in the sham surgery normal oxygen group ( P>0.05); it was 2.2(2, 2.5)mm in the ASDH normal oxygen group, significantly higher than that in the sham surgery normal oxygen group and sham surgery hypoxia group ( P<0.01), it was 3.1(2.9, 3.2)mm in the ASDH hypoxia group, significantly higher than that in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group ( P<0.01); it was 2.8(2.7, 2.9)mm in the ASDH hypoxia+oxygen inhalation group, not statistically different from that in the ASDH hypoxia group ( P>0.05), but significantly increased compared with that in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group ( P<0.01). Before modeling, before craniotomy and after craniotomy, the microvascular blood flow was 224.2±49.7, 224.8±50.3, 225.1±50.3 respectively in the sham surgery normal oxygen group and 224.7±43.7, 220.9±45.9, 221.8±45.5 respectively in the sham surgery hypoxia group, with no significant difference between the two groups ( P>0.05); it was 226.5±52.7, 173.4±40.7, 172.0±40.7 respectively in the ASDH normal oxygen group, significantly decreased compared with that in the sham surgery normal oxygen group and sham surgery hypoxia group ( P<0.05); it was 225.7±46.4, 131.4±23.6 and 131.0±23.5 respectively in the ASDH hypoxia group, significantly decreased compared with that in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group ( P<0.05); it was 226.2±56.1, 132.6±21.7 and 131.7±21.9 respectively in ASDH hypoxia+oxygen inhalation group, significantly decreased compared with that in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group ( P<0.05), with no significant difference from that in the ASDH hypoxia group ( P>0.05). At 0, 30 and 60 minutes after modeling, the expression levels of α-SMA and PDGFR-β were 0.70±0.02, 0.67±0.01, 0.55±0.05 and 0.65±0.03, 0.56±0.03 and 0.59±0.02 respectively in the sham surgery normal oxygen group and were 0.63±0.04, 0.60±0.01 0.55±0.05 and 0.62±0.01, 0.51±0.01 and 0.60±0.02 respectively in the sham surgery hypoxia group, with no significant difference between the two groups ( P>0.05); they were 0.88±0.06, 0.87±0.05, 0.82±0.03 and 0.85±0.03, 0.85±0.03, 0.88±0.04 respectively in the ASDH normal oxygen group, significantly higher than those in the sham surgery normal oxygen group and sham surgery hypoxia group ( P<0.01); they were 1.19±0.08, 1.10±0.10, 0.97±0.04 and 1.04±0.06, 1.19±0.07, 1.27±0.08 respectively in the ASDH hypoxia group, significantly higher than those in sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group ( P<0.05 or 0.01); they were 1.20±0.07, 1.10±0.04, 0.96±0.04 and 1.04±0.05, 1.15±0.11, 1.20±0.07 respectively in ASDH hypoxia+oxygen inhalation group, significantly higher than those in sham surgery normal oxygen group, sham surgery normal group and ASDH normal oxygen group ( P<0.01), but with no significant difference from those in ASDH hypoxia group ( P>0.05). At 0 minute after modeling, the fluorescence expression of α-SMA and PDGFR-β was weaker in the sham surgery normal oxygen group and the fluorescence expression of CD31 was stronger. There was no significant difference in the fluorescence expressions of α-SMA, PDGFR-β and CD31 between the sham surgery hypoxia group and sham surgery normal oxygen group. The fluorescence expressions of α-SMA and PDGFR-β in the ASDH normal oxygen group were stronger than those in the sham surgery normal oxygen group and sham surgery hypoxia group, while the fluorescence expression of CD31 was weaker. The fluorescence expressions of α-SMA and PDGFR-β in ASDH hypoxia group were stronger than those in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group, while the fluorescence expression of CD31 was weaker. The fluorescence expressions of α-SMA and PDGFR-β in the ASDH hypoxia+oxygen inhalation group were stronger than those in the sham surgery normal oxygen group, sham surgery hypoxia group and ASDH normal oxygen group, while the fluorescence expression of CD31 was weaker, with no significant difference from the fluorescence expressions of α-SMA, PDGFR-β and CD31 in ASDH hypoxia group. Conclusions:Hypoxia in ASDH rats will stimulate pericytes contraction, which causes cerebral microcirculatory disturbance, thus leading to TBS. Short-term inhalation of oxygen of medium concentration cannot dilate pericytes or microcirculation vessels, with no obvious effect on improving the conditions of TBS.

Through reviewing the ancient and modern literature， the name， origin， producing area， quality evaluation， harvesting and processing methods of Trichosanthis Fructus（TF） and Trichosanthis Radix（TR） in famous classical formulas were systematically sorted out following the chronological order. The results showed that there were many nicknames of TF and TR， and Gualou and Tianhuafen have become the mainstream names for its fruit and root， respectively. Both of them took Trichosanthes kirilowii as the mainstream base. TF and TR have been used as medicines in the Han dynasty， and since the North and South dynasties， Leigong Paozhilun had been clear that the effects of peels， seeds， stems， roots were different. TF was used as medicine with intact fruits， harvested after maturity from September to October， hung and dried in the shade， and its quality has been summarized in recent times as being best for those who are mature， large， thick and pliable peels， orange-yellow in color， and with sufficient sugary properties. In ancient times， the processing of TR was mostly crushed or shredded with the peels and seeds， or processing for pancakes and creams. TR was used as medicine with the roots， it is harvested from November to December， peeled and dried in the sun， and its quality was best when it was deep in the soil， large， white， powdery， firm and delicate with few muscles and veins， and it was considered to be toxic when it was born in briney land. Processing method of TR was to do powder into the medicine in the Tang dynasty， and gradually evolved into direct slicing use in the Ming and Qing dynasties. Since the modern era， the authentic producing areas of TF and TR were in the vicinity of Lingbao， Henan province， known as Anyang Huafen， and in modern times， there are well-known production areas such as Anguo， which produces Qihuafen， and Jinan， which produces Changqing Gualou. In the Song dynasty， there was a habit of substituting Trichosanthis Semen for the whole herb， which was later corrected by the materia medica in Ming dynasty. Based on the results， It is suggested that T. kirilowii be selected as the basal plant for the development of famous classical formulas involving TR and TF. In Qingjin Huatantang， Trichosanthis Semen is processed by stir-frying method， while TR and TF in other five formulas from the Catalogue of Ancient Famous Classical Formulas（The First Batch） were all used in raw form.