ObjectiveTo investigate the inhibitory effect of Biejiajian Pills （BJJW） on the growth of liver cancer， as well as its potential mechanism in mediating the AMP-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） pathway through mitochondrial energy metabolism. MethodsHuman hepatoma Huh7 cells were used to establish a nude mouse model of subcutaneous xenograft tumor. A total of 18 tumor-bearing nude mice were randomly divided into model group， BJJW group （2.2 g/kg）， and metformin group （250 mg/kg）， and the corresponding drug was given by gavage for 14 consecutive days. Tumor volume and weight were monitored during the experiment； HE staining was used to observe histopathological changes； the levels of reactive oxygen species （ROS） and adenosine triphosphate （ATP） in tumor tissue were measured； immunohistochemistry and Western blotting were used to measure the expression levels of proteins associated with the AMPK/mTOR pathway. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the Tukey’s test was used for further comparison between two groups； the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups， and the Dunn’s test was used for further comparison between two groups. ResultsCompared with the model group， the BJJW group had a tumor inhibition rate of 45.73%， with significant reductions in both tumor volume and weight （P<0.01）. Pathological examination showed that compared with the model group， the BJJW group had a significant reduction in the number of tumor cells and the presence of extensive necrosis. Mechanistic studies showed that compared with the model group， the BJJW group had a significant increase in ROS level （P<0.001） and a significant reduction in ATP level （P<0.001）， as well as significant increases in p-AMPK/AMPK ratio （0.81±0.20 vs 0.13±0.04， P<0.01） and p-ULK1/ULK1 ratio （0.69±0.17 vs 0.18±0.13， P<0.01） and a significant reduction in p-mTOR/mTOR ratio （1.34±0.16 vs 3.20±0.62， P<0.01）. ConclusionBJJW may inhibit the growth of liver cancer by inducing mitochondrial energy metabolism dysfunction， increasing the level of ROS， reducing the level of ATP， and activating the AMPK/mTOR signaling pathway.

Objective: Patients with atherosclerotic cardiovascular disease (ASCVD) following percutaneous coronary intervention (PCI) are classified as very-high-risk individuals in cardiovascular disease (CVD) risk stratification. The distribution pattern of traditional Chinese medicine (TCM) syndromes in this patient population, as well as its association with blood lipid profiles and clinical prognosis, remains unclear. The present prospective cohort study aims to investigate these correlations, thereby providing insights to enrich the research fields. Methods: We enrolled consecutive patients with ASCVD who underwent PCI at the Integrated Cardiology Unit of China-Japan Friendship Hospital between September 1, 2020 and December 31, 2022. Demographics and clinical characteristics, signs and symptoms defining each TCM syndrome, and fasting venous blood samples were collected at baseline and follow up or upon major adverse cardiovascular events (MACEs). We analyzed the correlation between TCM syndromes, blood lipid profiles, and MACEs, and developed a new joint prognostic model incorporating both TCM syndromes and blood lipids using logistic regression. The analyses were based on detailed baseline and one-year follow-up data. Results: A per-protocol analysis was performed on 586 patients with complete data ultimately. During the one-year follow-up, 174 patients (29.69%) experienced a MACE. We performed statistical analyses on comorbidities, medication, and biochemical indicators across groups defined by TCM syndrome differentiation. When comparing different TCM syndromes, no significant differences were found in age, body mass index (BMI), history of revascularization, comorbidities, family history of CVD, smoking or drinking, or statin intensity (P > 0.05). Patients with intertwined phlegm and blood stasis syndrome exhibited significantly higher levels of total cholesterol (TC, 5.27 ± 1.18 mmol/L, P < 0.001), triglyceride (TG, 1.96 ± 1.33 mmol/L, P = 0.008), low-density lipoprotein cholesterol (LDL-C, 3.35 ± 0.79 mmol/L, P < 0.001), and high-density lipoprotein cholesterol (HDL-C, 1.24 ± 0.81 mmol/L, P < 0.001) compared with those with other TCM syndromes combined. A multivariable logistic regression model was constructed to predict MACEs. The model included TCM syndrome type [with intertwined phlegm and blood stasis as a predictor, adjusted odds ratio (OR) = 1.413, 95% confidence interval (CI): 0.517 – 3.864, P = 0.501], age (adjusted OR = 0.97, 95% CI: 0.955 – 1.001, P = 0.057), male gender (adjusted OR = 0.698, 95% CI: 0.416 – 1.170, P = 0.173), TC (adjusted OR = 1.004, 95% CI: 0.513 – 1.965, P = 0.990), and LDL-C (adjusted OR = 5.825, 95% CI: 2.214 – 15.326, P < 0.001). This model demonstrated good discriminatory ability for MACEs in post-PCI ASCVD patients [the area under the receiver operating characteristic (ROC) curve (AUC) = 0.865, 95% CI: 0.816 – 0.914]. Conclusion The intertwined phlegm and blood stasis TCM syndrome is associated with a distinct atherogenic lipid profile characterized by elevated levels of TC and LDL-C. The prognostic model that incorporates this TCM syndrome type along with conventional lipid parameters (TC and LDL-C) shows good discriminatory ability for predicting MACEs in ASCVD patients after PCI, underscoring the potential clinical utility of integrating TCM syndrome differentiation into CVD risk assessment.

Objective: To explore the association between meat consumption and anxiety symptoms in first year junior high school students in Yunnan Province, and to provide theoretical support for preventing and relieving anxiety symptoms in junior high school students. Methods: From October to December 2022, a random cluster sampling method was used to select 8 500 first year junior high school students from 11 counties in Yunnan Province as the survey subjects for a questionnaire survey. The study used Food Frequency Questionnaire and the Chinese version of the Depression Anxiety Stress Scale-21 (DASS-21) to assess the meat consumption and anxiety symptoms of junior high school students.The distribution differences in anxiety symptoms among first year junior high school students with different demographic characteristics were analyzed statistically by using the Chi-square test,and the association between meat consumption and anxiety symptoms in students was analyzed by using a generalized linear model. Results: The detection rate of anxiety symptoms was 48.47%. After controlling for demographic variables and confounding factors, the consumption of livestock meat, poultry meat, processed meat, cured meat, barbecued meat and raw skin meat was statistically significant with anxiety symptoms ( β =-0.05, 0.04, 0.04, 0.08, 0.14, 0.17, all P <0.05). Stratified by ethnicity, The consumption of livestock meat, cured meat and barbecue was statistically correlated with anxiety symptoms in Han adolescents ( β =-0.07, 0.14, 0.22 ); the consumption of processed meat and raw skin meat was statistically correlated with anxiety symptoms in ethnic minority adolescents ( β =0.08, 0.18) (all P <0.05). Conclusions There is a statistical association between meat comsumption and the risk of anxiety symptoms in first year junior high school students in Yunnan Province. Guidance on meat consumption should be strengthened to prevent the occurrence of anxiety symptoms.

To investigate the risk factors for acute kidney injury (AKI) following the use of amphotericin B deoxycholate and to develop a predictive model to guide clinical monitoring and intervention.

OBJECTIVE To investigate the neuroprotective effect and mechanism of eleutheroside B （ELB） on Parkinson’s disease （PD） model mice by regulating the IκB kinase β （IKKβ）/nuclear factor-κB （NF-κB） signaling pathway. METHODS Fifty mice were randomly divided into normal control group， model group， positive control group （selegiline hydrochloride， 10 mg/kg）， and ELB low-dose and high-dose groups （80， 160 mg/kg）， with 10 mice in each group. Each group was given relevant medicine or normal saline intragastrically for 14 consecutive days. Starting from the 10th day of administration， the model group and all administration groups were intraperitoneally injected with 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine （MPTP） 30 mg/kg， for five consecutive days to establish the chronic PD model. After the last administration for 24 h， six mice were randomly selected from each group to test their behavioral abilities； detect the levels of interleukin-1β （IL-1β）， IL-10， tumor necrosis factor-α （TNF-α） in brain tissue and their mRNA expressions were measured， and positive expression of tyrosine hydroxylase （TH）， protein expressions of TH， α -synuclein （ α -syn）， ionized calcium-binding adaptor molecule 1 （Iba-1）， as well as phosphorylation levels of IKKβ and NF-κB p65 proteins in the brain tissue were detected. The ultrastructure of neurons in substantia nigra was observed. RESULTS Compared with the model group， rotarod endurance time and climbing score of each administration group （except for the ELB low-dose group） were increased significantly （ P ＜0.05）， while the levels and mRNA expressions of IL-1β， TNF-α， α -syn， and Iba-1， as well as phosphorylation levels of IKKβ and NF-κB p65 proteins in brain tissue were decreased significantly （except for TNF-α in the ELB low-dose group）. Conversely， the level and mRNA expression of IL-10 （except for the ELB low-dose group）， TH positive expression and protein expressions were significantly increased （ P ＜0.05）. Typical neurodegenerative pathological changes， such as neuronal karyopyknosis， mitochondrial swelling and vacuolization， and endoplasmic reticulum dilation， all showed varying degrees of improvement. CONCLUSIONS ELB may exert neuroprotective effects by inhibiting the activation of the IKKβ/NF-κB signaling pathway， alleviating inflammatory responses， reducing abnormal α -syn aggregation and neuronal loss， and further improving motor dysfunction in PD mice.

ObjectiveTo investigate the ability of clinically isolated， Helicobacter pylori （H. pylori）-specific gene polymerase chain reaction （PCR）-positive gastric， vaginal， and fecal Candida to release H. pylori. MethodsResuscitate 4 strains of H. pylori -specific 16S rDNA and ureA gene PCR-positive Candida strains isolated in laboratory from clinical sources， including 1 strain of gastric Candida， 1 strain of fecal Candida， 2 strains of vaginal Candida and the standard Candida albicans strain ATCC10231 （Ca10231）. The presence of H. pylori-specific ureA in the 5 strains of Candida isolates was confirmed by PCR. The aforementioned strains of Candida and H.pylori were inoculated into urea medium and cultured in a constant temperature incubator at 37 ℃. The color change of the medium was observed daily. A change in the medium's color from yellow to red indicated the presence of urease activity. Then， the five strains of Candida and H. pylori were co-incubated with the magnetic beads coated with H. pylori antibodies respectively. Scanning electron microscopy （SEM） was employed to observe the presence of bacilli adsorbed on the surface of the magnetic beads. PCR was used to detect the presence of H.pylori-specific 16S rDNA and ureA genes on magnetic beads. ResultsThe PCR analysis of the ureA gene in the four Candida isolates was positive， whereas the Ca10231 strain tested negative. Upon culturing the four Candida isolates on urea medium， the medium color changed from yellow to red which was determined to be urease positive， while the medium containing Ca10231 remained unchanged， which was urease negative. SEM revealed that bacilli could be observed on the surface of magnetic beads co-incubated with the 4 strains of Candida of clinical origin and H.pylori isolate. Specifically， PCR testing of the magnetic beads co-incubated with one vaginal Candida， one gastric Candida and H.pylori isolate showed positive results for the 16S rDNA and ureA genes of H. pylori； however， the PCR tests for the two genes were negative for the magnetic beads co-incubated with the other two Candida isolate. ConclusionThis study demonstrates that H. pylori-specific genes Candida can release H. pylori.

AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Objective:To evaluate the therapeutic efficacy of the Wenyang Huazhuo Formula on spontaneous pregnancy outcomes in advanced-age infertile patients with spleen-kidney yang deficiency syndrome.Methods:A retrospective cohort study design was employed to gather clinical data from 303 advanced-age infertile patients with spleen-kidney yang deficiency syndrome who sought treatment at the Department of Reproductive Medicine in the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine between June 2019 and June 2023. Patients were categorized into two groups based on their treatment regiments: a basic treatment group (non-exposed group, n=151) and a group receiving basic treatment in conjunction with the Wenyang Huazhuo Formula (exposed group, n=152). The study compared and analyzed pregnancy outcomes, including time to pregnancy (TTP) within one year, ongoing pregnancy rate, and clinical pregnancy rate, between the two groups over a one-year follow-up period. Additionally, stratified analyses were performed based on age and anti-Müllerian hormone (AMH) levels to evaluate the impact of Wenyang Huazhuo Formula on pregnancy outcomes across different patient subgroups. Results:The TTP in the exposed group was significantly reduced compared with the non-exposed group ( HR=1.70, 95% CI: 1.12-2.57, P=0.013), suggesting a significant shortening of their TTP, and the ongoing pregnancy rate [28.29% (43/152)] and the clinical pregnancy rate [33.55% (51/152)] were significantly higher than those in the non-exposed group [13.91% (21/151), P=0.020; 21.85% (33/151), P=0.023], and the early abortion rate [15.69% (8/51)] was significantly reduced compared with the non-exposed group [36.36% (12/33), P=0.031]. Stratified analyses by age revealed that for patients aged 35-37 years, the exposed group exhibited a significantly shorter TTP ( HR=1.67, 95% CI: 1.03-2.70, P=0.038), while no significant difference in TTP was observed in the 38-44-year old patients ( HR=1.78, 95% CI: 0.77-4.10, P=0.174). Regarding the ongoing pregnancy rate, the exposed group showed a significant higher rate [31.31% (31/99)] than the non-exposed group [13.91% (15/112), P=0.022] among patients aged 35-37 years, whereas no significant difference was observed in the 38-44 years old group ( P=0.386). Stratified analysis based on AMH levels showed that both low AMH levels (<1.68 μg/L) and high AMH levels (≥1.68 μg/L) patients in the exposed group experienced a significant reduction in TTP compared with the non-exposed group ( HR=8.07, 95% CI: 1.04-62.52, P=0.046; HR=1.57, 95% CI: 1.01-2.43, P=0.044, respectively), along with a notable improvement in the ongoing pregnancy rate [13.64% (6/44) vs. 0% (0/29), P=0.038; 34.26% (37/108) vs.17.21% (21/122), P=0.028, respectively]. Further stratified analysis indicated that, compared with patients with low AMH levels, both the exposed group and the non-exposed group of patients with high AMH levels exhibited a significantly shorter TTP ( HR=1.91, 95% CI: 1.02-3.69, P=0.043; HR=9.48, 95% CI: 1.30-69.22, P=0.002, respectively), along with a significantly higher ongoing pregnancy rate ( P=0.014; P=0.008). Conclusion:The Wenyang Huazhuo Formula significantly shortens TTP, enhances both the clinical pregnancy rate and the ongoing pregnancy rate and reduces the early abortion rate in advanced-age infertile women diagnosed with spleen-kidney yang deficiency syndrome.

Objective:To study on the relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus.Methods:120 elderly patients with GDM were treated in Obstetrics Department of Yantai Yuhuangding Hospital from Jun. 2019 to Jun. 2021. The healthy women (58 cases) and normal pregnant women (58 cases) were selected as the control group at the same time, and the clinical data and serum samples of subjects were collected. The levels of serum SC, SV, FPG and HbAlc were measured, the HOMA-IR of the aged GDM was calculated, and the difference of serum index expression among the subjects in each group was analyzed. The relationship between serum SC, SV and insulin resistance was analyzed by Person correlation, and the risk factors of GDM were analyzed by Logistic regression.Results:: There was significant difference in HbAlc, HOMA-IR, FPG, SC, and SV among the three groups. The levels of each index in the elderly GDM group were significantly higher than those in the normal pregnancy group and the healthygroup ( P<0.05). Pearson analysis showed that SC ( P=0.558, 0.626, 0.672), SV ( P=0.576, 0.663, 0.696) was positively correlated with HbAlc, HOMA-IR, FPG in elderly patients with GDM ( P<0.001). Logistic regression analysis showed that SC, SV was risk factors for GDM in elderly pregnant women. Conclusion:There is a positive correlation between serum SC, SV level and insulin resistance, which are risk factors for GDM in elderly pregnant women.

This study was aimed at investigating the presence of Helicobacter hepaticus(Hh)infection in patients with digestive tract diseases and evaluating Helicobacter pylori(Hp)infection status in patients with digestive tract cancers other than gastric cancer.Fecal samples were collected from 197 patients with digestive tract diseases at the Affiliated Cancer Hospital of Guizhou Medical Uni-versity and from 149 healthy volunteers residing in Guiyang.Hp stool antigen(HpSA)was detected with the colloidal gold method.Af-ter the extraction of fecal DNA,the Hp specific ureA gene and the Hh specific 16S rRNA gene were amplified via nested PCR,and the amplified products were subsequently confirmed through sequencing analysis.The study included 197 patients with digestive system diseases,comprising 135 cases of colorectal cancer,32 cases of chronic gastritis,22 cases of gastric cancer,5 cases of liver cancer,and 3 cases of cholangiocarcinoma.The detection rate of HpSA was 31.5%(62/197).HpSA was detected across all five disease catego-ries,and the highest detection rate was observed in patients with gastric cancer,at 50.0%(11/22),or colorectal cancer,at 24.4%(33/135).The positivity rate of Hp ureA gene PCR was 7.6%(15/197),and sequencing confirmed that the amplified products were in-deed Hp ureA gene fragments.Notably,the highest detection rate was observed in patients with colorectal cancer,at 8.9%(12/135).The positivity rate of Hh 16S rRNA gene PCR was 11.2%(22/197),and sequencing confirmed that the amplified products were in-deed Hh 16S rRNA gene fragments.Hh 16S rRNAgene presence was detected in patients with all five diseases,and the highest detec-tion rate was observed in patients with colorectal cancer,at 11.1%(15/135).Among 149 healthy volunteers,the detection rate of HpSA was 11.4%(17/149),only one case tested positive for the Hp ureA gene,and the Hh 16S rRNA gene was undetectable in all samples.In conclusion,Hh infection was detected in patients with digestive tract diseases.Beyond patients with gastric cancer,the prevalence of Hp infection was also notably high among patients with colorectal cancer,liver cancer,and cholangiocarcinoma.Further investigation is warranted to elucidate the roles of the two species of Helicobacter in the occurrence and progression of digestive tract cancers.