ObjectiveTo investigate the mechanisms by which the combination of berberine （BBR） and baicalin （BAI） ameliorates type 2 diabetes mellitus （T2DM） due to internal accumulation of dampness-heat from the perspectives of gut microbiota and metabolomics. MethodsAntibiotics were used to induce pseudo-sterile mice. Thirty pseudo-sterile mice were randomized into a normal fecal microbiota transplantation group （n=10） and a T2DM （syndrome of internal accumulation of dampness-heat） fecal microbiota transplantation group （n=20）. The mice were then administrated with suspensions of fecal microbiota from healthy volunteers and a patient with T2DM due to internal accumulation of dampness-heat by gavage， respectively. Each mouse received 200 µL suspension every other day for a total of 15 times to reshape the gut microbiota. The T2DM model mice were then assigned into a model group （n=8） and a BBR-BAI group （n=11）. BBR was administrated at a dose of 200 mg·kg^-1， and BAI was administrated in a ratio of BBR-BAI 10∶1 based on preliminary research findings. The administration lasted for 8 consecutive weeks. Fasting blood glucose （FBG）， glycated hemoglobin （HbA1c）， insulin （INS）， triglycerides （TG）， total cholesterol （CHOL）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） levels were measured to evaluate the effects of the BBR-BAI combination on glucose and lipid metabolism and liver function in T2DM mice. Hematoxylin-eosin staining was employed to observe pathological changes in the colon tissue. The expression of claudin-1， zonula occludens-1 （ZO-1）， and occludin in the colon tissue was determined by Western blot. Real-time quantitative polymerase chain reaction（Real-time PCR） was employed to assess the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the colon tissue. The fecal microbiota composition and differential metabolites were analyzed by 16S rRNA sequencing and ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry （UPLC-Q-TOF-MS）， respectively. ResultsThe BBR-BAI combination lowered the FBG， HbA1c， and INS levels （P<0.05， P<0.01） and alleviated insulin resistance （P<0.01） in T2DM mice. Additionally， BBR-BAI elevated the levels of ZO-1， occludin， and claudin-1 （P<0.05， P<0.01） and down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in the colon （P<0.05， P<0.01）. The results of 16S rRNA sequencing showed that BBR-BAI increased the relative abundance of Ligilactobacillus， Phascolarctobacterium， and Akkermansia （P<0.05）， while significantly decreasing the relative abundance of Alistipes， Odoribacter， and Colidextribacter （P<0.05）. UPLC-Q-TOF-MS identified 28 differential metabolites， which were primarily involved in arachidonic acid metabolism and α-linolenic acid metabolism. ConclusionBBR-BAI can ameliorate T2DM due to internal accumulation of dampness-heat by modulating the relative abundance of various bacterial genera in the gut microbiota and the expression of fecal metabolites.

Objective To explore the value of inflammatory load index(IBI)in the diagnosis and prognosis of gastric cancer patients.Methods Clinical data of patients who underwent gastric cancer surgery in the Department of General Surgery of the Second Hospital of Lanzhou University from January 2016 to December 2018 were retrospectively analyzed,and the patients were divided into the high IBI group and the low IBI group based on the optimal cut-off value of IBI.ROC curve was used to determine the optimal cut-off value of IBI,and the survival curve was constructed using the Kaplan-Meier method,and the differences between groups were examined using the Log-rank test.Cox regression analysis was performed to study the prognostic factors of gastric cancer patients.Results The best cut-off value for IBI diagnosis was 8.796,and the best cut-off value for prognosis was 28.75.IBI was related to the clinical case characteristics of gastric cancer patients,such as surgical access,the degree of differentiation,CK7,CK20,LMP-1,and Ki-67(P＜0.05).The results of univariate analysis showed that intraoperative bleeding,preoperative CEA,preoperative CA125,preoperative CA199,IBI,tumor diameter size,margins of incision,vascular invasion,nerve invasion,pT stage and pN stagewere the influencing factors on the prognosis of gastric cancer patients(P＜0.05).The results of multifactorial regression analysis showed that intraoperative bleeding,preoperative CEA,preoperative CA199,IBI,tumor diameter size,pT stage and pN stagewere independent risk factors affecting the prognosis of gastric cancer patients(P＜0.05).The median OS of patients in the high IBI group and low IBI group were 32.9 months and 74.8 months,respectively,with statistically significant differences(P＜0.001).Conclusion IBI is an independent risk factor for poor prognosis of patients with gastric cancer,and it has a good predictive value in diagnosis and prognosis.

Objective To explore the value of inflammatory load index(IBI)in the diagnosis and prognosis of gastric cancer patients.Methods Clinical data of patients who underwent gastric cancer surgery in the Department of General Surgery of the Second Hospital of Lanzhou University from January 2016 to December 2018 were retrospectively analyzed,and the patients were divided into the high IBI group and the low IBI group based on the optimal cut-off value of IBI.ROC curve was used to determine the optimal cut-off value of IBI,and the survival curve was constructed using the Kaplan-Meier method,and the differences between groups were examined using the Log-rank test.Cox regression analysis was performed to study the prognostic factors of gastric cancer patients.Results The best cut-off value for IBI diagnosis was 8.796,and the best cut-off value for prognosis was 28.75.IBI was related to the clinical case characteristics of gastric cancer patients,such as surgical access,the degree of differentiation,CK7,CK20,LMP-1,and Ki-67(P＜0.05).The results of univariate analysis showed that intraoperative bleeding,preoperative CEA,preoperative CA125,preoperative CA199,IBI,tumor diameter size,margins of incision,vascular invasion,nerve invasion,pT stage and pN stagewere the influencing factors on the prognosis of gastric cancer patients(P＜0.05).The results of multifactorial regression analysis showed that intraoperative bleeding,preoperative CEA,preoperative CA199,IBI,tumor diameter size,pT stage and pN stagewere independent risk factors affecting the prognosis of gastric cancer patients(P＜0.05).The median OS of patients in the high IBI group and low IBI group were 32.9 months and 74.8 months,respectively,with statistically significant differences(P＜0.001).Conclusion IBI is an independent risk factor for poor prognosis of patients with gastric cancer,and it has a good predictive value in diagnosis and prognosis.

Rare diseases are a collective term for diseases with extremely low prevalence and incidence rates.Up to now,China has released two lists identifying a total of 207 rare diseases.Given that most rare diseases do not have drugs with corresponding indications,physicians frequently resort to using off-label drugs when treating patients with rare diseases.However,there is currently no systematic guideline or expert consensus for the use of off-label medications in China.To comprehensively collect existing evidence of off-label drug use for rare diseases,fully analyze and evaluate the rationality of off-label drug use for rare diseases,and standardize the management of off-label drug use for rare diseases,the Rare Disease Branch of Beijing Medical Association,Chinese Pharmaceutical Association,Beijing Pharmaceutical Association,and the School of Public Health,Peking University have jointly initiated the drafting of the Expert Consensus on Off-label Use of Drugs for Rare Diseases.This consensus refer to the WHO Handbook for Guideline Development,the Guidelines for Developing/Revising Clinical Diagnostic and Treatment Guidelines in China(2022 Edition),the AGREE Ⅱ and the STAR tools.This protocol outlines the background and purpose of consensus,as well as the comprehensive framework for consensus development,encompassing panel formation,clinical issue identification,evidence retrieval,data extraction,and evidence-based recommendation formulation.

Objective:To develop a health education intervention program for minor stroke patients based on the information-knowledge-attitude-practice (IKAP) theory and to verify its application effectiveness.Methods:A convenience sampling method was used to select 31 minor stroke patients admitted to the Emergency Department of Xuanwu Hospital, Capital Medical University, from September to October 2023 as the control group, and another 31 patients admitted from November to December 2023 as the experimental group. The control group received routine health education, while the experimental group implemented the health education program based on the IKAP theory. Knowledge, attitude, and practice of the patients were assessed using the Stroke Knowledge, Attitude, and Practice Questionnaire before the intervention, immediately after the intervention, and 3 months post-intervention.Results:Immediately after the intervention, the knowledge, attitude, and behavior scores of patients in the experimental group were higher than those of the control group, and the differences were statistically significant (all P<0.05); three months after the intervention, the knowledge and attitude scores of patients in the experimental group were higher than those of the control group, and the differences were statistically significant (all P<0.05). Conclusions:The health education intervention program based on the IKAP theory can effectively improve the knowledge, attitude, and practice levels of minor stroke patients, with lasting effects.

To evaluate the immunogenicity and immunoprotective effects of a tRNA thiouridylase TgMnmA deletion strain of Toxoplasma gondii(T.gondii)on ICR mice,we constructed a mouse model immunized with RH△MnmA.Mice were immunized with 10 RH△MnmA tachyzoites by in-traperitoneal injection.After 30 d,indirect ELISA was used to detect the specific IgG antibody and its subtypes of immunized mice.Spleen lymphocyte suspension was prepared,and the splenic lym-phocyte subsets were analyzed by flow cytometry.Moreover,the relative expression level of cyto-kine mRNA was detected by real-time fluorescence quantitative PCR.After 30 d of immunization,mice were intraperitoneally inoculated with RH△ku80 tachyzoites.At 5 d post infection,the para-site load in the ascites,heart,liver and brain of mice was measured,and the survival of mice within 30 d after infection was observed and recorded.The results showed that compared with the control PBS group,RH△MnmA immunized group produced higher level of IgG and IgG2a antibodies,higher mRNA relative expression level of cytokines IL-2,IL-4,IL-6,IL-10,IL-12 and IFN-γ,and the number of CD4+and CD8a+in spleen lymphocytes also increased significantly.Mean-while,for the attack of RH△ku80 strain,the immune group can effectively reduce the parasite load in the ascites and some tissues,inhibit the reproduction of parasites In vivo,and significantly improve the survival rate of mice.The results of this study showed that the TgMnmA deletion strain of T.gondii can induce strong humoral and cellular immune responses in mice,and provide good immune protection against the infection of RHΔku80 strain,which has the potential to be-come a potentially promising live attenuated vaccine candidate against T.gondii.

In recent years,with the constant rise in the number of infertility patients,the rapid and accurate assessment of sperm quality has become a key challenge in reproductive medicine.Currently,traditional methods for sperm classification suffer from limitations in efficiency,subjectivity,and cost-effectiveness,and hence fail to meet clinical needs.With the introduction of artificial intelligence(AI),innovative approaches have been offered to address these issues.Herein,we systematically reviewed the latest progress in AI applications in the field of male infertility,focusing on the role of AI in the assessment of sperm concentration,motility,morphology,DNA fragmentation index,and the treatment of non-obstructive azoospermia.It has been reported that AI technologies,such as convolutional neural networks,demonstrate high accuracy and efficiency in assessing sperm concentration and motility.Particularly in morphological analysis,the performance of AI has been validated in multiple studies,significantly enhancing objectivity and clinical utility.However,the assessment of DNA fragmentation index remains underexplored due to the lack of support in advanced imaging technology,with only a few models showing promise.Additionally,AI significantly improves sperm detection rates in modified testicular sperm extraction through AI-driven image recognition,offering a breakthrough in the treatment of patients with non-obstructive azoospermia.We also discussed the application of natural language processing technology in patient pre-consultation and follow-up,such as automated data collection and intelligent tracking systems,which demonstrate AI's potential to optimize medical workflows.In the future,with the accumulation of high-quality datasets,algorithm optimization,and advances in imaging technology,the application of artificial intelligence is expected to enable multi-dimensional comprehensive screening and play a greater role in the diagnosis and treatment of male infertility.

Objective To investigate the effects of body weight and length of SD rats on the carotid artery balloon injury-induced vascular stenosis model in order to provide a reference for replicating an ideal vascular stenosis mod-el.Methods Male rats were divided into three groups based on body weight and length.The CONQUEROR? SC PTCA balloon catheter was employed,with a fixed balloon inflation volume of 0.2 mL to induce injury in the left common carotid artery,while the right side served as a control.As soon as surgery operation,one rat from each group was selected for Evans Blue dye verification.Fourteen days later,the injured and contra lateral common ca-rotid arteries from remaining rats were harvested for HE staining to check the extent of stenosis.Based on these find-ings,six rats within the optimal range of body weight and length were selected for further validation.Results Rats with body weights ranging from 280 to 380 g(corresponding body lengths of 21.0-26.5 cm)underwent balloon catheter injury,resulting in endothelial detachment and varying degrees of stenosis in the common carotid artery.In rats weighing 280-300 g(body lengths of 21.0-22.5 cm)had severe stenosis or occlusion of the common carotid artery with thrombosis.In rats weighing 320-340 g(body lengths of 23.0-24.5 cm),the internal and external elastic plates of the common carotid artery were ruptured and the vascular morphology was abnormal.Conversely,rats weighing 360-380 g(body lengths of 25.0-26.5 cm)did not show any ruptured elastic laminae or thrombus formation in the common carotid artery,and the extent of vascular stenos in rats with a body weight of 360 g was moderate and uniform.The results of the repeated validation experiments were consistent.Conclusions Rats with a body weight range of 360 g(corresponding body length of 25.0-26.5 cm)are suitable for development of an ideal vascular stenosis model.

ObjectiveTo observe the clinical efficacy of Gandou Fumu Granules （GDFMG） combined with sodium dimercaptosulphonate （DMPS） on liver fibrosis in Wilson disease （WD） patients with the syndrome of phlegm and blood stasis， evaluate its effect on cuproptosis-related indicators， and explore the possible mechanisms of cuproptosis in WD-related liver fibrosis. MethodsSixty WD patients diagnosed with the syndrome of phlegm and blood stasis between January 2023 and December 2023 were randomly divided into a control group and an observation group， with 30 patients in each group. The control group received the copper chelator DMPS for the first 6 days， followed by calcium gluconate injection for the next 2 days， completing an 8-day treatment cycle. The observation group received GDFMG in addition to the treatment regimen of the control group， with both groups treated for 21 cycles. A Beckman fully automated biochemical analyzer was used to detect levels of type Ⅳ collagen （CⅣ）， hyaluronic acid （HA）， laminin （LN）， N-terminal propeptide of type Ⅲ procollagen （PⅢ-NP）， and serum copper （SCu） before and after treatment in both groups. Enzyme-linked immunosorbent assay （ELISA） was used to measure levels of ferredoxin 1 （FDX1）， lipoic acid synthetase （LIAS）， and dihydrolipoamide S-acetyltransferase （DLAT）. Atomic absorption spectroscopy measured 24-hour urine copper levels before treatment and after the 7， 14， and 21 treatment cycles in both groups. An Fibro Touch （FT） non-invasive liver fibrosis diagnostic device was used to measure liver stiffness （LSM） in both groups before and after treatment. Traditional Chinese medicine syndrome score （TCMSS） was evaluated at the same intervals. Clinical efficacy， adverse events， and safety indicators were also compared. ResultsAfter treatment， levels of CⅣ， HA， LN， and PⅢNP significantly decreased in both groups compared to pre-treatment levels （P<0.01）. The observation group showed a more pronounced reduction compared to the control group （P<0.05）. There were no statistically significant differences in SCu levels in both groups before and after treatment. After treatment， levels of FDX1，LIAS and DLAT significantly increased in both groups（P<0.01）. The observation group showed more notable improvements in these indicators than the control group （P<0.05）. After the 7， 14， 21 treatment cycles， 24-hour urine copper levels significantly increased in both groups compared to pre-treatment levels （P<0.01）. The observation group had a greater increase in 24-hour urine copper levels than the control group after treatment （P<0.05，P<0.01）， and although 24-hour urine copper levels increased after 7 cycles， a gradual decline was observed in subsequent cycles. After treatment， LSM levels significantly decreased in both groups compared to pre-treatment levels （P<0.01）， with the observation group showing a greater reduction than the control group （P<0.05）. Clinical efficacy was significantly better in the observation group than the control group （P<0.05）. No significant differences in the incidence of adverse events or safety indicators were observed between the two groups after treatment. ConclusionGDFMG combined with DMPS can reduce LSM in WD patients with liver fibrosis and the syndrome of phlegm and blood stasis， inhibit cuproptosis， and improve clinical efficacy.

To evaluate the immunogenicity and immunoprotective effects of a tRNA thiouridylase TgMnmA deletion strain of Toxoplasma gondii(T.gondii)on ICR mice,we constructed a mouse model immunized with RH△MnmA.Mice were immunized with 10 RH△MnmA tachyzoites by in-traperitoneal injection.After 30 d,indirect ELISA was used to detect the specific IgG antibody and its subtypes of immunized mice.Spleen lymphocyte suspension was prepared,and the splenic lym-phocyte subsets were analyzed by flow cytometry.Moreover,the relative expression level of cyto-kine mRNA was detected by real-time fluorescence quantitative PCR.After 30 d of immunization,mice were intraperitoneally inoculated with RH△ku80 tachyzoites.At 5 d post infection,the para-site load in the ascites,heart,liver and brain of mice was measured,and the survival of mice within 30 d after infection was observed and recorded.The results showed that compared with the control PBS group,RH△MnmA immunized group produced higher level of IgG and IgG2a antibodies,higher mRNA relative expression level of cytokines IL-2,IL-4,IL-6,IL-10,IL-12 and IFN-γ,and the number of CD4+and CD8a+in spleen lymphocytes also increased significantly.Mean-while,for the attack of RH△ku80 strain,the immune group can effectively reduce the parasite load in the ascites and some tissues,inhibit the reproduction of parasites In vivo,and significantly improve the survival rate of mice.The results of this study showed that the TgMnmA deletion strain of T.gondii can induce strong humoral and cellular immune responses in mice,and provide good immune protection against the infection of RHΔku80 strain,which has the potential to be-come a potentially promising live attenuated vaccine candidate against T.gondii.