Objective To detect the expression of lactate dehydrogenase A(LDHA),FoxP3,and CD4 in hepatocellular carcinoma(HCC)and to analyze the effect of bufalin on LDHA as well as on the differentiation and function of regulatory T(Treg)cells in the tumor microenvironment of HCC mice.Methods Immunohistochemical staining was performed to detect the difference in the expression of LDHA,FoxP3(Treg cell marker),and CD4 between the tumor and adjacent tissues from 91 HCC patients.Hepl-6 subcutaneous tumor-bearing mouse model was established.The mice were randomly divided into control,bufalin+pyruvic acid,oxalate,and bufalin groups(n=5 each group).The mass and volume of subcutaneous tumors were compared.Immunohistochemical staining was performed to detect the expression of LDHA,FoxP3,and CD4 in tumors of mice in each group.ELISA was used to detect the levels of transforming growth factor β(TGF-β)and interleukin-10(IL-10)in the serum of mice in each group to evaluate the immune function of Treg cells.Results Compared with adjacent tissues,HCC tissues exhibited significantly higher LDHA and FoxP3 expression and lower CD4 expression(P＜0.001).The results of the correlation analysis showed that LDHA was positively correlated with FoxP3 expression and negatively correlated with CD4 expression(P＜0.05).The experimental results of Hep1-6 subcutaneous tumor-bearing mice showed that the tumor volume was significantly smaller in the bufalin and oxalate groups than in the control group and was significantly larger in the bufalin+pyruvic acid group than in the bufalin group(P＜0.001).The results of immunohistochemical staining showed that the expression levels of LDHA and the percentage of FoxP3+cells in the bufalin and oxalate groups were significantly lower than those in the control group,and the percentage of CD4+cells in the bufalin group was significantly higher than that in the control group(P＜0.001).The expres-sion level of LDHA and the percentage of FoxP3+cells in the bufalin+pyruvic acid group were significantly higher than those in the bufalin group,while the percentage of CD4+cells was significantly lower than that in the bufalin group(P＜0.001).The results of ELISA showed that the levels of TGF-β and IL-10 in the bufalin and oxalate groups were significantly lower than those in the control group(P＜0.001).Conclusion Bufalin inhibits cell growth,downregulates LDHA expression,and suppresses the differentiation and proliferation of Treg cells in HCC mice.

Hepatocellular carcinoma(HCC),as the most common type of liver cancer,poses a significant threat to global public health due to its high incidence and mortality rates.This paper delves into the etiology,pathogenesis,and syndrome differentiation of liver cancer from the perspective of"dispersing qi and fortifying the body resistance",based on the clinical experience of renowned traditional Chinese medicine(TCM)practitioner,Prof.Changquan Ling.Prof.Ling believes that the development of liver cancer is closely related to the disruption of liver qi flow,the accumulation of blood stasis over time,and the generation of toxin from long-term stagnation,accompanied by pathological changes such as imbalance of yin and yang,deficiency of the body's vital qi and accumulation of pathogenic factors,and internal blazing of cancer toxins.In terms of treatment,he emphasizes the principles of dispersing qi and fortifying the body resistance,addressing both the root cause and symptoms.This is achieved by regulating the functions of viscera,improving the stagnation of qi flow,and supplemented by methods such as clearing heat and detoxifying,and softening and dispersing hard masses,aiming to break the vicious cycle of qi stagnation,deficiency of vital qi,and pathogenic factor generation,thereby promoting the recovery from the disease.Through detailed analysis of clinical cases,this paper demonstrates Prof.Ling's unique insights and significant efficacy in treating liver cancer through"dispersing qi"to"fortify the body resistance",ultimately achieving"tumor suppression".This provides new references and perspectives for the clinical diagnosis and treatment of liver cancer in TCM.

Hepatocellular carcinoma(HCC),as the most common type of liver cancer,poses a significant threat to global public health due to its high incidence and mortality rates.This paper delves into the etiology,pathogenesis,and syndrome differentiation of liver cancer from the perspective of"dispersing qi and fortifying the body resistance",based on the clinical experience of renowned traditional Chinese medicine(TCM)practitioner,Prof.Changquan Ling.Prof.Ling believes that the development of liver cancer is closely related to the disruption of liver qi flow,the accumulation of blood stasis over time,and the generation of toxin from long-term stagnation,accompanied by pathological changes such as imbalance of yin and yang,deficiency of the body's vital qi and accumulation of pathogenic factors,and internal blazing of cancer toxins.In terms of treatment,he emphasizes the principles of dispersing qi and fortifying the body resistance,addressing both the root cause and symptoms.This is achieved by regulating the functions of viscera,improving the stagnation of qi flow,and supplemented by methods such as clearing heat and detoxifying,and softening and dispersing hard masses,aiming to break the vicious cycle of qi stagnation,deficiency of vital qi,and pathogenic factor generation,thereby promoting the recovery from the disease.Through detailed analysis of clinical cases,this paper demonstrates Prof.Ling's unique insights and significant efficacy in treating liver cancer through"dispersing qi"to"fortify the body resistance",ultimately achieving"tumor suppression".This provides new references and perspectives for the clinical diagnosis and treatment of liver cancer in TCM.

Objective To investigate the efficacy and safety of conventional open Latarjet surgery,arthroscopic Latarjet surgery,arthroscopic Bankart repair combined with Remplissage surgery in the treatment of recurrent anterior shoulder dislocation(RASD)with scapula glenoid bone defect>15%and meshing Hill-Sachs lesion.Methods The clinical data of 65 patients with RASD with 15%～25%scapula glenoid bone defect and meshing Hill-Sachs lesion admitted to our hospital from January 2022 to December 2024 were retrospectively analyzed.They were divided into group A,group B,and group C according to different surgical methods.Among them,group A underwent conventional open Latarjet surgery(n=18),group B underwent arthroscopic Latarjet surgery(n=21),and group C underwent arthroscopic Bankart repair combined with Remplissage surgery(n=26).The surgical conditions,preoperative and postoperative scores of related scales[visual analogue scale(VAS),Constant-Murley shoulder score,the University of California Los Angeles(UCLA)shoulder score],shoulder range of motion,postoperative complications and recurrence were compared among the three groups.Results Operation time:group B was longer than group A and group C(P<0.05),and group A was longer than group C(P<0.05).Intraoperative blood loss and hospital stay:group A was more or longer than group B and group C(P<0.05),group B was more or longer than group C(P<0.05).The VAS scores of the three groups at 1,6 and 12 months after operation were lower than those before operation(P<0.05).Pain VAS score at 1 and 6 months after operation:group A was higher than group B and group C(P<0.05).Pain VAS score at 1 month after operation:group B was higher than group C.Pain VAS score at 12 months after operation:there was no significant difference among the three groups(P>0.05).The UCLA shoulder scores of group A at 6 and 12 months after operation and group B and group C at 1,6 and 12 months after operation were higher than those before operation(P<0.05).UCLA score at 1 month after operation:group A0.05).The Constant-Murley scores of group A at 6 and 12 months after operation and group B and group C at 1,6 and 12 months after operation were higher than those before operation(P<0.05).Constant-Murley score at 1 month after operation:group A was lower than group B and group C(P<0.05).Constant-Murley score at 6 and 12 months after operation:three groups were equivalent(P>0.05).Forward flexion and lifting range of motion in the three groups:at 12 months after operation>before operation(P<0.05).Forward flexion and lifting range of motion was evaluated at 12 months after operation:there was no significant difference among the three groups(P>0.05).The range of motion of external rotation at body side and external rotation at 90° abduction before operation and at 12 months after operation:there was no significant difference between group A and group B(P>0.05).External rotation at body side and external rotation at 90° abduction of group C:at 12 months after operation of it was less than that before operation(P<0.05).The range of motion of external rotation at body side and external rotation at 90° abduction was obtained at 12 months after operation:group A and group B were greater than group C(P<0.05).Postoperative complication rate and recurrence rate:three groups were equivalent(P>0.05).Conclusion Conventional open Latarjet surgery,arthroscopic Latarjet surgery,arthroscopic Bankart repair combined with Remplissage surgery can effectively improve shoulder joint function and relieve pain in the treatment of RASD with 15%～25%scapula glenoid bone defect and meshing Hill-Sachs lesion.However,the two shoulder arthroscopic procedures are less traumatic and have faster postoperative recovery.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective To detect the expression of lactate dehydrogenase A(LDHA),FoxP3,and CD4 in hepatocellular carcinoma(HCC)and to analyze the effect of bufalin on LDHA as well as on the differentiation and function of regulatory T(Treg)cells in the tumor microenvironment of HCC mice.Methods Immunohistochemical staining was performed to detect the difference in the expression of LDHA,FoxP3(Treg cell marker),and CD4 between the tumor and adjacent tissues from 91 HCC patients.Hepl-6 subcutaneous tumor-bearing mouse model was established.The mice were randomly divided into control,bufalin+pyruvic acid,oxalate,and bufalin groups(n=5 each group).The mass and volume of subcutaneous tumors were compared.Immunohistochemical staining was performed to detect the expression of LDHA,FoxP3,and CD4 in tumors of mice in each group.ELISA was used to detect the levels of transforming growth factor β(TGF-β)and interleukin-10(IL-10)in the serum of mice in each group to evaluate the immune function of Treg cells.Results Compared with adjacent tissues,HCC tissues exhibited significantly higher LDHA and FoxP3 expression and lower CD4 expression(P＜0.001).The results of the correlation analysis showed that LDHA was positively correlated with FoxP3 expression and negatively correlated with CD4 expression(P＜0.05).The experimental results of Hep1-6 subcutaneous tumor-bearing mice showed that the tumor volume was significantly smaller in the bufalin and oxalate groups than in the control group and was significantly larger in the bufalin+pyruvic acid group than in the bufalin group(P＜0.001).The results of immunohistochemical staining showed that the expression levels of LDHA and the percentage of FoxP3+cells in the bufalin and oxalate groups were significantly lower than those in the control group,and the percentage of CD4+cells in the bufalin group was significantly higher than that in the control group(P＜0.001).The expres-sion level of LDHA and the percentage of FoxP3+cells in the bufalin+pyruvic acid group were significantly higher than those in the bufalin group,while the percentage of CD4+cells was significantly lower than that in the bufalin group(P＜0.001).The results of ELISA showed that the levels of TGF-β and IL-10 in the bufalin and oxalate groups were significantly lower than those in the control group(P＜0.001).Conclusion Bufalin inhibits cell growth,downregulates LDHA expression,and suppresses the differentiation and proliferation of Treg cells in HCC mice.

OBJECTIVE To investigate the effects of triptolide(TP)exposure during pregnancy and lactation on the reproductive system development and function in male offspring of rats,providing a reference for medication safety during pregnancy and lactation.METHODS Pregnant rats were randomly divided into control group(12 rats,normal saline)and T1-T4 groups[12,13,14,17 rats that received TP at 200,400,600,and 800 μg/(kg·d)respectively].They were given relevant medicine/normal saline intragastrically,once a day,until the offspring were born and naturally weaned,the intragastric administration volume of each rat was consistently 2 mL.After 60 days of feeding,reproductive organ weights and coefficients were measured in male offspring,testicular and epididymal histology and sperm morphology were observed.Sperm motility,sperm count,and serum levels of gonadotropin-releasing hormone(GnRH),follicle stimulating hormone(FSH),luteinizing hormone(LH),and testosterone(T)in the epididymides were analyzed.Protein expressions of glycogen synthase kinase 3α(GSK3α),phosphorylated GSK3α(p-GSK3α),and phosphatase 1γ2(PP1γ2)in sperm were also determined.RESULTS Compared with the control group,the testicular and epididymal weights,serum levels of GnRH and T,the relative protein expression of PP1γ2 were significantly decreased in T1-T4 groups.Additionally,in the T2 to T4 groups,there were significant reductions in the weight and coefficient of the seminal vesicle,total number of sperm,sperm concentration,sperm motility as well as relative protein expressions of GSKα,p-GSK3α in the offspring rats.Furthermore,the epididymal coefficient in the T3 and T4 groups,the testicular coefficient,mean sperm track velocity and sperm curvature velocity in the T4 group were significantly decreased(P＜0.05);the number of abnormal sperm,rate of sperm abnormality,and levels of FSH and LH in the offspring rats of the T1 to T4 groups were all significantly increased(P＜0.05);in the offspring rats of the T1 to T4 groups,there was a decrease in the number of epithelial cells in the seminiferous tubules of the testes.Within the epididymal tissue,degenerative and necrotic changes in the epithelial cells were visible,accompanied by mild infiltration of inflammatory cells in the stroma.CONCLUSIONS TP exposure during pregnancy and lactation disrupts reproductive organ development,impairs spermatogenesis and sperm motility,as well as suppresses androgen synthesis in male offspring,thereby having a negative impact on the development of the reproductive system.These effects may be mechanistically linked to regulation of GSK3α,p-GSK3α and PP1γ2 protein expressions.

Objective To investigate the diagnostic value of dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI)combined with Doppler ultrasound and elastography in breast lesions.Methods A total of 91 patients with breast lesions confirmed by pathology were retrospectively selected and underwent DCE-MRI,Doppler ultrasound and elastography examinations.The imaging features of benign and malignant breast lesions were analyzed.The diagnostic value of DCE-MRI,Doppler ultrasound,and elastogra-phy in benign and malignant breast lesions were analyzed.Results Among the 91 cases,there were 62 cases with benign lesions(benign group)and 29 cases with malignant lesions(malignant group).There were statistically significant differences in DCE-MRI parameters,Doppler ultrasound parameters and elastography parameters between the two groups(P<0.05).Binary logistic regres-sion analysis found that enhancement characteristics,morphology,time-signal intensity curve(TIC),blood flow signals on Doppler ultrasound,elastography score,and strain rate(SR)were the influencing factors of benign and malignant breast lesions(P<0.05).The differential diagnostic efficacy of combination of the three examinations was highest.The sensitivity,specificity and accuracy were 88.71%,82.76%and 86.81%,respectively.Conclusion DCE-MRI,Doppler ultrasound,and elastography are effective for differentiating benign and malignant breast lesions.Combination of them can improve the diagnostic performance.

Objective:To explore the value of blood lactic acid (BLA), procalcitonin (PCT), and total bilirubin (TBil) in the diagnosis and prognosis evaluation of patients with trauma complicated with sepsis.Methods:The clinical data of 151 patients with severe trauma admitted to the Department of Emergency Medicine, Nanxishan Hospital of Guangxi Zhuang Autonomous Region from July 2019 to August 2023 were analyzed retrospectively. The patients were divided into the sepsis group (72 cases) and non-sepsis group (79 cases) according to the diagnosis. They were further divided into the death group (37 cases) and non-death group (114 cases) based on clinical outcomes. Clinical data were compared between groups. Receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of the above indicators, and Spearman correlation analysis was applied to evaluate the correlation between the indicators.Results:The levels of BLA, PCT, TBil, and Sequential Organ Failure Assessment (SOFA) score in the sepsis group were higher than those in the non-sepsis group (all P<0.05). The mortality rate of the sepsis group was significantly higher than that of the non-sepsis group, with a statistically significant difference [26/72(36.11%) vs 11/79(13.92%), χ 2=10.024, P=0.002]. The levels of BLA, PCT, TBil, and SOFA score in the death group were higher than those in the non-death group (all P<0.05). ROC curve analysis showed that the areas under the curve (AUC) of BLA, PCT, TBil, and their combination for diagnosing sepsis were 0.745, 0.826, 0.753, and 0.889 respectively; the sensitivity and specificity of the combined diagnosis of sepsis were 87.5% and 72.2%. The AUCs of BLA, PCT, TBil, and their combination for predicting the prognosis of sepsis were 0.644, 0.697, 0.614, and 0.713 respectively; the sensitivity and specificity of the combined prediction of sepsis prognosis were 64.9% and 71.1%. Among the 151 patients, the levels of BLA, PCT, TBil were positively correlated with SOFA score, with statistically significant differences ( r=0.3871, 0.4399, 0.4851, all P<0.001). Conclusions:BLA, PCT, and TBil levels have certain value in the early diagnosis and prognosis evaluation of patients with sepsis. The combined evaluation has the best efficacy and high guiding value in clinical practice.