Objective To elucidate the changes in the gut microbiota and molecular mechanism of huaier in enhancing the efficacy of oxaliplatin (OXA) chemotherapy in gastric cancer (GC). Methods The effects of huaier on the gut microbiota structure and intestinal barrier function in MKN45-bearing mice were analyzed by using 16S rRNA sequencing, RT-qPCR, and IHC. UPLC-MS and network pharmacology, as well as in vivo experimental approaches, were employed to investigate the key bioactive constituents of huaier systematically and elucidate the underlying mechanisms by which huaier exerts therapeutic effects against GC. The expression of the PI3K/AKT/SREBP pathway was detected in cancer cells and tissues via Western blot analysis. The safety profile of huaier combined with OXA was verified through serum biochemistry and HE-stained tissue section analyses. Results Huaier inhibited the PI3K/AKT/SREBP pathway by increasing the abundance of Akkermansia muciniphila, reduced lipid droplet deposition, and ultimately enhanced the sensitivity to OXA in the treatment of GC. Conclusion This study demonstrates the value of huaier in gastric cancer treatment by enhancing chemosensitivity and provides novel insights into its systemic therapeutic effects through molecular mechanisms and gut microbiota modulation.

Objective:To explore the mechanism of Shiwei Chaihu Shugan Powder in the treatment of breast hyperplasia using network pharmacology; To verify the mechanism of Shiwei Chaihu Shugan Powder in the treatment of breast hyperplasia through animal experiments.Methods:The active components and potential targets of Shiwei Chaihu Shugan Powder were searched in TCMSP and Uniprot databases. Breast hyperplasia genes were searched in GeneCards and OMIM databases. The intersection targets were obtained by online tool Venny, and the "drug-component-target" network was constructed by Cytoscape 3.8.2 software. The protein interaction (PPI) network was constructed using the String platform, and GO function and KEGG pathway enrichment analysis were performed using the DAVID annotation database. Molecular docking was performed using PDB, PubChem database, PyMOL 2.1 and AutoDockvina 1.2.5 software to predict the biological mechanism of Shiwei Chaihu Shugan Powder in the treatment of breast hyperplasia. Rats were divided into blank group, model group, tamoxifen group and Shiwei Chaihu Shugan Powder low-, medium- and high-dosage groups according to the random number table method, with 6 rats in each group. Except for the blank group, the other groups were prepared with the modified estrogen-progesterone-induced rat mammary hyperplasia model. Shiwei Chaihu Shugan Powder low-, medium- and high-dosage groups were intragastrically administered with Shiwei Chaihu Shugan Powder solution at 7.425 g/kg, 14.850 g/kg, and 29.700 g/kg respectively, while the tamoxifen group was intragastrically administered with 2.1 mg/kg tamoxifen. The blank group and the model group were intragastrically administered with the same volume of drinking water, once a day, for consecutive 28 d. The thickness of the mammary gland was measured by small animal ultrasound. The height and width of the nipples were measured by vernier calipers. The levels of serum E2 and P were detected by ELISA. The morphology of mammary tissue was observed by HE staining. The expressions of ERα, ERβ, SRC-1 and CBP/p300 proteins were detected by Western blot.Results:A total of 92 active components and 274 disease-drug intersection targets were screened out. GO functional enrichment analysis showed that Shiwei Chaihu Shugan Powder was closely related to positive regulation of gene expression, positive regulation of RNA polymerase Ⅱ promoter transcription, signal transduction, negative regulation of apoptosis process, response to heterogeneous stimulation, and regulation of hormone levels. KEGG enrichment analysis showed that the core targets might be related to NF-κB signaling pathway, MAPK signaling pathway, AGE-RAGE signaling pathway, PI3K/Akt signaling pathway, and regulating hormone levels. Molecular docking results showed that the core components had a good binding energy with the core target and a stable conformation. Compared with the model group, the thickness of the mammary gland in the tamoxifen group and Shiwei Chaihu Shugan Powder low-, medium- and high-dosage groups decreased ( P<0.01), the serum P level increased ( P<0.05), the expressions of ERα, SRC-1, and CBP/p300 proteins decreased ( P<0.01), and the expression of ERβ protein increased ( P<0.01); the height of the nipples in the Shiwei Chaihu Shugan Powder medium- and high-dosage groups and the tamoxifen group decreased ( P<0.01), and the serum E2 level increased ( P<0.05). Conclusion:Shiwei Chaihu Shugan Powder may play a role in the treatment of breast hyperplasia by regulating the levels of estrogen and related proteins.

OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with Osteopathia striata with cranial sclerosis (OSCS) due to variant of AMER1 gene. METHODS: A child presented at the Affiliated Children's Hospital of Zhengzhou University in July 2024 due to growth and development retardation was selected as the study subject. A retrospective study was conducted to collect the child's clinical data. Peripheral blood samples (2 mL each) were collected from the child and her parents, and genomic DNA was extracted for whole exome sequencing (WES). Sanger sequencing was used for the verification of candidate variants. The pathogenicity of variant was rated according to the guidelines from American College of Medical Genetics and Genomics (ACMG). The study has been approved by the Medical Ethics Committee of the Children's Hospital Affiliated to Zhengzhou University (Ethics No.: 2024-108-001). RESULTS: The patient, a 4-year-and-10-month-old girl, presented with global developmental delay, short stature, cleft palate, distinct facial features, and hearing impairment. WES revealed that she has harbored a heterozygous c.790_794dup (p.Cys265Trpfs*19) variant of the AMER1 gene, which was not detected in either parent. Based on the guidelines from ACMG, the gene variant was classified as pathogenic (PVS1 + PS2 + PM2_supporting). As the result of a non-triplet base insertion in the coding region of the AMER1 gene, it has converted a codon originally encoding an amino acid into a stop codon, and led to a truncated protein, causing severe alteration and dysfunction of the protein. CONCLUSION The child was diagnosed with OSCS for clinical features such as global developmental delay, short stature, cleft palate, distinctive facial features, and hearing impairment, for which the de novo heterozygous frameshift variant AMER1: c.790_794dup (p.Cys265Trpfs*19) may be accountable. Above finding has expanded the mutational spectrum of OSCS and provided a basis for genetic counseling and prenatal diagnosis for the family.
Humans ; Female ; Child, Preschool ; Osteosclerosis/genetics* ; Adaptor Proteins, Signal Transducing/genetics* ; Mutation ; Exome Sequencing ; Retrospective Studies ; Tumor Suppressor Proteins

Objective:To explore the effect of psoas muscle index (PMI) and construct a machine learning model to validate the 180-day prognosis in patients with decompensated liver cirrhosis.Methods:Retrospective data were collected from patients with decompensated liver cirrhosis at Henan Provincial People's Hospital from January 2022 to November 2022. The area of the psoas muscle index (PMI) at the level of the third lumbar vertebra was measured and calculated based on the abdominal X-ray computed tomography images stored in the Eastern China Hospital Information System (HIS). Patients were divided into low PMI and normal PMI groups according to the receiver operating characteristic curve. Patients clinical data and complication status were collected.The general conditions of both groups were compared using a t-test, chi-square test, and Mann-Whitney U test. The Kaplan-Meier method was applied for survival analysis. The outcome variable was 180-day mortality, and variables were selected using Cox and LASSO regression. The dataset was divided into training and testing sets in a 7∶3 ratio. Machine learning algorithms were used to build models in the training set, and model performance was validated by the test set. The model for MELD-Na score was compared with the model for End-Stage Liver Disease score. Results:A total of 298 patients with decompensated liver cirrhosis were included.The MELD scores, Child-Pugh classification, and NRS2002 scores, along with the incidence rate of complications such as ascites, hepatic encephalopathy, infections, and gastrointestinal bleeding, were significantly higher in the low PMI than the normal PMI group, with statistically significant differences ( P<0.05). The area under a receiver operating characteristic curve for the extreme gradient boosting model was higher than traditional clinical scores (MELD score 0.658, MELD_Na score 0.719) in the machine learning model. Furthermore, the application of SHAP results model indicated that PMI, hemoglobin, NRS2002 score, direct bilirubin, and blood ammonia were important factors in predicting the prognosis of patients with decompensated liver cirrhosis. Conclusion:A low PMI is closely related to poorer survival rates and the development of complication rates in patients with decompensated liver cirrhosis. The machine learning prediction model based on this construction, especially extreme gradient boosting, has favorable predictive performance, which is superior to the traditional clinical scoring system and can provide patients with the most accurate risk assessment and individualized treatment plan.

Objective:To explore the clinical characteristics and genetic etiology of a child with Osteopathia striata with cranial sclerosis (OSCS) due to variant of AMER1 gene. Methods:A child presented at the Affiliated Children′s Hospital of Zhengzhou University in July 2024 due to growth and development retardation was selected as the study subject. A retrospective study was conducted to collect the child′s clinical data. Peripheral blood samples (2 mL each) were collected from the child and her parents, and genomic DNA was extracted for whole exome sequencing (WES). Sanger sequencing was used for the verification of candidate variants. The pathogenicity of variant was rated according to the guidelines from American College of Medical Genetics and Genomics (ACMG). The study has been approved by the Medical Ethics Committee of the Children′s Hospital Affiliated to Zhengzhou University (Ethics No.: 2024-108-001).Results:The patient, a 4-year-and-10-month-old girl, presented with global developmental delay, short stature, cleft palate, distinct facial features, and hearing impairment. WES revealed that she has harbored a heterozygous c. 790_794dup (p.Cys265Trpfs*19) variant of the AMER1 gene, which was not detected in either parent. Based on the guidelines from ACMG, the gene variant was classified as pathogenic (PVS1 + PS2 + PM2_supporting). As the result of a non-triplet base insertion in the coding region of the AMER1 gene, it has converted a codon originally encoding an amino acid into a stop codon, and led to a truncated protein, causing severe alteration and dysfunction of the protein. Conclusion:The child was diagnosed with OSCS for clinical features such as global developmental delay, short stature, cleft palate, distinctive facial features, and hearing impairment, for which the de novo heterozygous frameshift variant AMER1: c. 790_794dup (p.Cys265Trpfs*19) may be accountable. Above finding has expanded the mutational spectrum of OSCS and provided a basis for genetic counseling and prenatal diagnosis for the family.

Objective:To study the effect of traditional Chinese medicine (TCM) formula in the treatment of brucellosis.Methods:Patients with brucellosis who were treated at the Beidahuang Industry Group General Hospital from March to November 2024 were selected and their clinical data were collected. A case-control study was conducted, thirty patients treated with conventional therapy plus TCM formula were selected as the TCM group, and 35 patients treated with conventional therapy were selected as the control group. Blood routine, C-reactive protein (CRP), lymphocyte subsets (CD45 +, CD3 +, CD4 +, CD8 +, CD19 +), and related cytokines [interleukin (IL)-6, IL-10] were determined before and after treatment to observe the clinical effect of TCM formula in the treatment of brucellosis. Survival curve was draw, and Log-Rank test was used to compare the differences in clinical symptom relief time between the two groups of patients. Results:Compared with pre-treatment, there were statistically significant differences in the numbers of CD45 +, CD3 +, CD4 +, CD8 +, CD19 + lymphocytes, neutrophil (NEUT), and the levels of CRP, IL-6, and IL-10 in the TCM group after treatment ( P < 0.05). After treatment, the comparison of each index between the two groups showed that there were statistically significant differences in the numbers of CD45 +, CD3 +, CD4 +, and CD8 + lymphocytes [control group vs TCM group: 2 470.00 (1 895.50, 3 207.00) vs 1 991.00 (1 720.75, 2 367.25), 1 920.00 (1 364.50, 2 428.00) vs 1 591.00 (1 343.00, 1 884.00), 1 021.00 (785.00, 1 205.50) vs 839.50 (704.25, 1 010.25), (686.42 ± 294.47) vs (596.97 ± 205.32) pieces/μl, P < 0.05]. There was no statistically significant difference in the number of CD19 + lymphocytes, NEUT, and the levels of CRP, IL-6 and IL-10 ( P > 0.05). The Log-Rank test results showed that there were statistically significant differences in the relief time of hyperhidrosis and night sweats ( P = 0.016), fatigue ( P = 0.016), and muscle soreness ( P = 0.004) between the two groups of patients. Conclusion:TCM formula has certain effects in the adjuvant therapy of brucellosis, which can improve the immune function of lymphocytes and relieve clinical symptoms, and has clinical application value.

Objective:To learn about the levels of 25-hydroxy vitamin D (25-OH VD), TBNK lymphocyte subsets, and cytokines in peripheral blood of patients with brucellosis.Methods:A prospective design was adopted, one hundred patients with brucellosis admitted to the Department of Infectious Diseases, Beidahuang Industry Group General Hospital from May 2024 to February 2025 were selected as the brucellosis group, and one hundred healthy individuals who underwent physical examinations at the hospital during the same period were selected as the control group. The peripheral blood 25-OH VD levels were detected by chemiluminescence method. Further, 100 patients with brucellosis were divided into a brucellosis combined with osteoarthritis group (74 cases) and a brucellosis without osteoarthritis group (26 cases). Flow cytometry was used to detect the counts of peripheral blood TBNK lymphocyte subsets and cytokine levels. Meanwhile, Spearman rank correlation was used to analyze the correlation between peripheral blood 25-OH VD levels and TBNK lymphocyte subsets counts as well as cytokine levels in patients with brucellosis complicated by osteoarthritis.Results:The peripheral blood 25-OH VD level in the brucellosis group [20.31 (15.74, 24.35) ng/ml] was significantly lower than that of the control group [25.18 (21.13, 29.59) ng/ml], and the difference was statistically significant ( Z = - 5.07, P < 0.001). The peripheral blood 25-OH VD level [18.05 (13.79, 23.74) vs 22.43 (19.93, 28.25) ng/ml], CD4 + T cell count [(860 ± 275) vs (1 036 ± 376) cells/μl], and interleukin (IL)-6 levels [4.17 (2.14, 9.41) vs 7.83 (5.97, 11.34) ng/L] in the brucellosis combined with osteoarthritis group were significantly lower than those in the brucellosis without osteoarthritis group ( Z/t = - 2.88, 2.20, - 2.85, P = 0.004, 0.035, 0.004). Correlation analysis showed that the peripheral blood 25-OH VD level in patients with brucellosis complicated by osteoarthritis was positively correlated with the counts of CD45 +, CD3 + T, CD4 + T, CD8 + T, and natural killer cells ( r = 0.31, 0.26, 0.25, 0.25, 0.25, P = 0.007, 0.027, 0.032, 0.031, 0.032), and negatively correlated with IL-17A level ( r = - 0.40, P < 0.001). Conclusion:Patients with brucellosis have insufficient 25-OH VD, and those with osteoarthritis have lower 25-OH VD level, CD4 + T cell count, and IL-6 level than those without osteoarthritis.

AIM:To study the mechanism of oxy-sophoridine(OSR)in relieving neuropathic pain(NP).METHODS:The analgesic effect of OSR was observed by measuring mechanical pain sensitivity.By combining OSR with agonists and antagonists re-lated to synthesis and metabolism of gamma ami-nobutyric acid(GABA),the mechanism related to analgesic effects of OSR and GABA nervous system is studied.The expression of c-Fos immunopositive cells and the expression of c-Fos and Glutamic acid decarboxylase(Glutamic acid decarboxylase 67)in brain and spinal cord were detected by immunoflu-orescence staining.Co-expression of GAD67,GABA transporter 1(gamma-Aminobutyric acid Transport-er 1,GAT-1)immunopositive cells.RESULTS:Com-pared with Sham group,spared nerve injury(SNI)group showed increased mechanical pain sensitivi-ty,increased expression of c-Fos immunopositive cells,decreased and increased co-expression of c-Fos and GAD67 and GAT-1 immunopositive cells,re-spectively.Compared with SNI group,mechanical algesia in OSR 500 and 1 000 mg/kg groups was de-creased,algesia in OSR 250 mg/kg combined with antagonist group was decreased,and algesia in OSR 500 mg/kg combined with agonist group was increased.The co-expression of c-Fos and GAD67 immunopositive cells in the brain and spinal cord of OSR 500 mg/kg group increased,while the co-ex-pression of c-Fos and GAT-1 decreased.CONCLU-SION:OSR has a good analgesic effect on NP mice induced by SNI.The mechanism is that OSR increas-es the content of central GABA by up-regulating GABAergic neurons in brain and spinal cord.

Objective:To investigate the application in evaluating the course and the clinical effects of serum cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) levels, as well as neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) in patients with brucellosis.Methods:Using case-control method, from February 2023 to March 2024, 274 confirmed brucellosis patients [divided into acute and chronic groups ( n = 165, 109) according to the course of the disease] and 70 healthy individuals (control group) were selected at Beidahuang Group General Hospital for serum cytokines detection using cytometric bead array (CBA) method. Blood routine test, serum agglutination test (SAT) and blood culture were performed at the same time, and NLR and PLR were calculated. Cytokine levels, NLR, and PLR were compared in patients with different disease duration, with or without complications, with different SAT titers [high ( > 1 ∶ 100) and low (≤1 ∶ 100)], and with different blood culture results, and the effects of each indicator on the course of brucellosis were analyzed by logistic regression. Results:The levels of IL-2, IL-6, IL-10, IL-17, TNF-α and IFN-γ in the acute group [ M ( Q1, Q3): 0.32 (0.15, 0.70), 18.97 (10.70, 36.86), 2.54 (1.49, 4.36), 1.41 (0.38, 3.05), 1.31 (0.77, 2.33), 11.60 (2.30, 36.75) ng/L] were higher than those in the chronic group [0.18 (0.06, 0.43), 1.68 (0.75, 5.74), 0.88 (0.40, 1.93), 0.29 (0.09, 0.87), 0.59 (0.31, 1.07), 0.72 (0.33, 1.42) ng/L] and control group [0.10 (0.05, 0.30), 1.52 (0.09, 2.80), 0.72 (0.35, 1.16), 0.08 (0.03, 0.20), 0.55 (0.20, 0.96), 0.68 (0.41, 1.25) ng/L, P < 0.05]. The IFN-γ level in the group with complications of brucellosis was lower than that in the group without complications, while the NLR and PLR were higher than those in the group without complications ( P < 0.05). The levels of IL-6, IL-17, TNF-α, and IFN-γ in the high titer group were higher than those in the low titer group, and the NLR was lower than that in the low titer group ( P < 0.05). The levels of IFN-γ and TNF-α of blood culture positive patients in the acute group were higher than those of blood culture negative patients ( P < 0.05). Univariate analysis showed that all 7 cytokines could affect the course of brucellosis ( P < 0.05). Multivariate analysis showed that IL-6, TNF-α, and IFN-γ were independent influencing factors of the course of brucellosis [ OR (95% CI) = 0.87 (0.83, 0.91), 0.55 (0.32, 0.97), 0.80 (0.72, 0.88), P < 0.05]. Conclusions:The levels of cytokines IL-2, IL-6, IL-10, IL-17, TNF-α, and IFN-γ can reflect the course progression of brucellosis patients, IL-6, IFN-γ and TNF-α can also serve as independent influencing factors for brucellosis progression. NLR and PLR may become inflammatory markers for predicting Brucella infection.

Objective:To analyze the risk factors of hypocalcemia in 3-5 stages of chronic kidney disease (CKD) patients with hyperkalemia and non-dialysis after potassium lowering therapy.Methods:Clinical data of 3-5 stages of CKD patients with hyperkalemia and non-dialysis treated in Linyi Central Hospital from January 2019 to November 2024 were collected through the electronic medical record system. According to whether the corrected calcium level after potassium lowering treatments was lower than 2.12 mmol/L, the patients were divided into hypocalcemia group and non-hypocalcemia group. The gender, age, body mass index, primary disease, disease duration, comorbidity, use of potassium lowering drugs, concomitant medication, and blood potassium, corrected calcium, carbon dioxide binding capacity, blood magnesium, blood phosphorus, estimated glomerular filtration rate, and total parathyroid hormone before potassium lowering treatments between the 2 groups were compared. Multiple logistic regression analysis was used to identify the risk factors for hypocalcemia in 3-5 stages of CKD patients with hyperkalemia and non-dialysis after potassium lowering therapy.Results:A total of 260 patients were entered, including 58 with blood calcium lower than 2.12 mmol/L, and incidence of hypocalcemia was 22.3%. The differences in the baseline corrected calcium, blood phosphorus, carbon dioxide binding capacity, estimated glomerular filtration rate, and total parathyroid hormone between the hypocalcemia group and the non-hypocalcemia group were statistically significant ( P<0.05). The factors with P<0.1, including primary disease, baseline corrected calcium, blood phosphorus, carbon dioxide binding capacity, estimated glomerular filtration rate, and total parathyroid hormone, were included in the multivariate logistic regression analysis. The results showed that the probability of hypocalcemia at baseline corrected calcium levels of 2.12-2.21, 2.22-2.31, and 2.32-2.41 mmol/L was 49.306 times, 13.651 times, and 13.342 times that of at ≥2.42 mmol/L, respectively. Low carbon dioxide binding capacity (odds ratio=0.909, 95% confidence interval: 0.836-0.987) was also a risk factor of hypocalcemia in 3-5 stages CKD patients with hyperkalemia and non-dialysis after potassium lowering therapy. Conclusions:Three to five stages of CKD patients with hyperkalemia and non-dialysis are prone to hypocalcemia after potassium lowering therapy. The low levels of baseline corrected calcium and carbon dioxide binding may be closely related to the occurrence of hypocalcemia in 3-5 stages of CKD patients with hyperkalemia and non-dialysis after potassium lowering therapy.