Traditional Chinese medicine （TCM） diagnostics is a discipline that studies the basic theories and fundamental skills of diagnostic methods， disease diagnosis， and differentiation in accordance with the theories of TCM. The artificial intelligence （AI） technology has gained remarkable achievements in the intelligentization of the four diagnostic methods in TCM and the standardization of differentiation and diagnosis. However， it still faces many challenges. The standardization of clinical data collection is difficult， and the data quality is uneven， which affects the usability of the data. The integration of the four diagnostic information is insufficient. Most instruments can only collect data from a single diagnostic method， lacking overall integrity. The scientific nature of the diagnostic model needs to be improved. The existing models lack dynamics and the reasoning logic of TCM differentiation. The accuracy of intelligent methods needs to be improved， and the existing evaluation indicators cannot fully reflect the practical application effect of the model. Furthermore， the relevant laws and regulations are still not perfect， and data security and patient privacy lack guarantees. The cultivation of compound talents is insufficient， and there is a lack of interdisciplinary talents who are proficient in both TCM and AI. On this basis， this paper expounded on the current development status， difficulties， and bottlenecks of AI in TCM diagnosis and then explored the development trend of AI in the field of TCM diagnosis. It proposed solutions such as optimizing the data collection process， constructing multimodal diagnostic models， facilitating multi-disciplinary exchanges and cooperation， improving laws and regulations， and cultivating compound talents. It is hoped that modern， standardized， normalized， and intelligent TCM diagnosis can be further promoted， thereby providing new impetus and methods for the inheritance and innovation of TCM.

OBJECTIVE: To analyze the Epstein-Barr virus (EBV) load in different lymphocyte subsets, as well as clinical characteristics and outcomes in patients with hematologic malignancies experiencing EBV reactivation. METHODS: Peripheral blood samples from patients were collected. B, T, and NK cells were isolated sorting with magnetic beads by flow cytometry. The EBV load in each subset was quantitated by real-time quantitative polymerase chain reaction (RT-qPCR). Clinical data were colleted from electronic medical records. Survival status was followed up through outpatient visits and telephone calls. Statistical analyses were performed using SPSS 25.0. RESULTS: A total of 39 patients with hematologic malignancies were included, among whom 35 patients had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median time to EBV reactivation was 4.8 months (range: 1.7-57.1 months) after allo-HSCT. EBV was detected in B, T, and NK cells in 20 patients, in B and T cells in 11 patients, and only in B cells in 4 patients. In the 35 patients, the median EBV load in B cells was 2.19×10⁴ copies/ml, significantly higher than that in T cells (4.00×10³ copies/ml, P <0.01) and NK cells (2.85×10² copies/ml, P <0.01). Rituximab (RTX) was administered for 32 patients, resulting in EBV negativity in 32 patients with a median time of 8 days (range: 2-39 days). Post-treatment analysis of 13 patients showed EBV were all negative in B, T, and NK cells. In the four non-transplant patients, the median time to EBV reactivation was 35 days (range: 1-328 days) after diagnosis of the primary disease. EBV was detected in one or two subsets of B, T, or NK cells, but not simultaneously in all three subsets. These patients received a combination chemotherapy targeting at the primary disease, with 3 patients achieving EBV negativity, and the median time to be negative was 40 days (range: 13-75 days). CONCLUSION In hematologic malignancy patients after allo-HSCT, EBV reactivation commonly involves B, T, and NK cells, with a significantly higher viral load in B cells compared to T and NK cells. Rituximab is effective for EBV clearance. In non-transplant patients, EBV reactivation is restricted to one or two lymphocyte subsets, and clearance is slower, highlighting the need for prompt anti-tumor therapy.
Humans ; Hematologic Neoplasms/virology* ; Herpesvirus 4, Human/physiology* ; Epstein-Barr Virus Infections ; Hematopoietic Stem Cell Transplantation ; Virus Activation ; Lymphocyte Subsets/virology* ; Flow Cytometry ; Killer Cells, Natural/virology* ; Male ; Female ; B-Lymphocytes/virology* ; Viral Load ; Adult ; T-Lymphocytes/virology* ; Middle Aged

Neonatal sepsis is a global health problem that seriously affects the body health and life safety of newborns. It has a higher incidence in preterm infants,especially for early-onset sepsis (EOS) within 72 hours of birth. The diagnosis of neonatal EOS requires a series of examinations,and early and accurate diagnosis can improve clinical outcomes and reduce antibiotic overuse in a timely manner. At present,the commonly used biomarkers and traditional blood culture methods for EOS diagnosis have certain shortcomings,so it is urgent to find new molecular diagnostic methods. This article summarizes and compares the early and novel diagnostic methods of neonatal EOS,in order to provide a reference for clinical practice.

Objective To explore the effects and therapeutic mechanism of electroacupuncture on the levels of polarization markers and inflammatory factors interleukin(IL)-6,IL-4,tumor necrosis factor-alpha(TNF-α),and IL-10 in rats with para-chlorophenylalanine-induced insomnia(PCPA).Methods Fifty healthy specific-pathogen free grade Sprague-Dawley rats,half male and half female,were randomly divided into a blank group(n=10)and a model reserve group(n=40),in which insomnia was induced by intraperitoneal injection of a 500 mg/kg PCPA suspension.Using the random number table method,the 30 successfully modeled rats were divided into three treatment groups of 10 rats/group:model,electroacupuncture,and estazolam.The estazolam group was given estazolam 0.2 mg/(kg·d)by gavage;the electroacupuncture group was given once-daily electroacupuncture at the"Shenmen"and"Sanyinjiao"acupoints,and stimulation at the"Baihui"and"Benshen"acupoints,20 minutes each time,for 7 consecutive days.Following treatment,serum and hypothalamic levels of TNF-α,IL-6,IL-4,and IL-10 were detected using ELISA and Western blot,while immunofluorescence staining was used to detect the presence of Iba-1 in hypothalamic microglia and the co-expression of CD86 and CD163,which are markers for the M1 and M2 subtypes of microglial cells,respectively.Results Compared with the blank group,the model group exhibited prolonged sleep latency(SL)(P＜0.01),shortened sleep duration(ST)(P＜0.05),significantly higher serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly lower levels of IL-4 and IL-10(P＜0.01).Compared with the model group,the electroacupuncture and estazolam groups exhibited significantly shorter SL(P＜0.01),prolonged ST(P＜0.01),significantly lower serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly higher IL-4 and IL-10 levels(P＜0.01).IL-6 content was lower in the electroacupuncture group than in the estazolam group(P＜0.05).Compared with the blank group,the model group exhibited significantly enhanced Iba-1/CD86(M1 type)co-expression(P＜0.01)alongside significantly weakened Iba-1/CD163(M2 type)co-expression(P＜0.01).Under electroacupuncture or estazolam intervention,Iba-1/CD86 co-expression was significantly weakened(P＜0.01),and Iba-1/CD163 co-expression was significantly enhanced in the model group(P＜0.05).Conclusions Electroacupuncture effectively improved sleep disturbances in rats,with an underlying mechanism that may involve regulation of microglial polarization,downregulation of pro-inflammatory cytokine levels,upregulation of anti-inflammatory cytokine levels,and alleviation of neuroinflammation,thereby ameliorating sleep.

Neonatal sepsis is a global health problem that seriously affects the body health and life safety of newborns. It has a higher incidence in preterm infants,especially for early-onset sepsis (EOS) within 72 hours of birth. The diagnosis of neonatal EOS requires a series of examinations,and early and accurate diagnosis can improve clinical outcomes and reduce antibiotic overuse in a timely manner. At present,the commonly used biomarkers and traditional blood culture methods for EOS diagnosis have certain shortcomings,so it is urgent to find new molecular diagnostic methods. This article summarizes and compares the early and novel diagnostic methods of neonatal EOS,in order to provide a reference for clinical practice.

Objective To explore the effects and therapeutic mechanism of electroacupuncture on the levels of polarization markers and inflammatory factors interleukin(IL)-6,IL-4,tumor necrosis factor-alpha(TNF-α),and IL-10 in rats with para-chlorophenylalanine-induced insomnia(PCPA).Methods Fifty healthy specific-pathogen free grade Sprague-Dawley rats,half male and half female,were randomly divided into a blank group(n=10)and a model reserve group(n=40),in which insomnia was induced by intraperitoneal injection of a 500 mg/kg PCPA suspension.Using the random number table method,the 30 successfully modeled rats were divided into three treatment groups of 10 rats/group:model,electroacupuncture,and estazolam.The estazolam group was given estazolam 0.2 mg/(kg·d)by gavage;the electroacupuncture group was given once-daily electroacupuncture at the"Shenmen"and"Sanyinjiao"acupoints,and stimulation at the"Baihui"and"Benshen"acupoints,20 minutes each time,for 7 consecutive days.Following treatment,serum and hypothalamic levels of TNF-α,IL-6,IL-4,and IL-10 were detected using ELISA and Western blot,while immunofluorescence staining was used to detect the presence of Iba-1 in hypothalamic microglia and the co-expression of CD86 and CD163,which are markers for the M1 and M2 subtypes of microglial cells,respectively.Results Compared with the blank group,the model group exhibited prolonged sleep latency(SL)(P＜0.01),shortened sleep duration(ST)(P＜0.05),significantly higher serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly lower levels of IL-4 and IL-10(P＜0.01).Compared with the model group,the electroacupuncture and estazolam groups exhibited significantly shorter SL(P＜0.01),prolonged ST(P＜0.01),significantly lower serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly higher IL-4 and IL-10 levels(P＜0.01).IL-6 content was lower in the electroacupuncture group than in the estazolam group(P＜0.05).Compared with the blank group,the model group exhibited significantly enhanced Iba-1/CD86(M1 type)co-expression(P＜0.01)alongside significantly weakened Iba-1/CD163(M2 type)co-expression(P＜0.01).Under electroacupuncture or estazolam intervention,Iba-1/CD86 co-expression was significantly weakened(P＜0.01),and Iba-1/CD163 co-expression was significantly enhanced in the model group(P＜0.05).Conclusions Electroacupuncture effectively improved sleep disturbances in rats,with an underlying mechanism that may involve regulation of microglial polarization,downregulation of pro-inflammatory cytokine levels,upregulation of anti-inflammatory cytokine levels,and alleviation of neuroinflammation,thereby ameliorating sleep.

Objective:To explore the characteristics of SLCO1B1/ SLCO1B3 gene variants among children with Rotor syndrome (RS). Methods:Four children who were admitted to the Department of Hepatology of Hunan Children′s Hospital between January 2019 and January 2022 were selected as the study subjects. Trio-whole exome sequencing was carried out for the four families, and gel electrophoresis was used to verify an insertional variant of long-interspersed element-1 (LINE-1).Results:Genetic testing has identified three variants of the SLCO1B1 gene, including c. 1738C>T (p.R580*), c. 757C>T (p.R253*) and c. 1622A>C (p.Q541P), and two variants of the SLCO1B3 gene, including c. 481+ 22insLINE-1 and c. 1747+ 1G>A among the children. Three of them were found to harbor homozygous variants of the SLCO1B1/ SLCO1B3 genes, and one has harbored compound heterozygous variants. Sanger sequencing confirmed the existence of all variants, and gel electrophoresis has confirmed the existence of the LINE-1 insertional variant of about 6 kb within intron 6 of the SLCO1B3 gene in all children. Conclusion:The pathogenesis of the RS among the four children may be attributed to the variants of the SLCO1B1/ SLCO1B3 genes. The LINE-1 insertion variant of the SLCO1B3 gene may be common among Chinese RS patients.

OBJECTIVES: To investigate the difference in intestinal microbiota between preterm infants with neurodevelopmental impairment (NDI) and those without NDI. METHODS: In this prospective cohort study, the preterm infants who were admitted to the neonatal intensive care unit of Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from September 1, 2019 to September 30, 2021 were enrolled as subjects. According to the assessment results of Gesell Developmental Scale at the corrected gestational age of 1.5-2 years, they were divided into two groups: normal (n=115) and NDI (n=100). Fecal samples were collected one day before discharge, one day before introducing solid food, and at the corrected gestational age of 1 year. High-throughput sequencing was used to compare the composition of intestinal microbiota between groups. RESULTS: Compared with the normal group, the NDI group had a significantly higher Shannon diversity index at the corrected gestational age of 1 year (P<0.05). The principal coordinate analysis showed a significant difference in the composition of intestinal microbiota between the two groups one day before introducing solid food and at the corrected gestational age of 1 year (P<0.05). Compared with the normal group, the NDI group had a significantly higher abundance of Bifidobacterium in the intestine at all three time points, a significantly higher abundance of Enterococcus one day before introducing solid food and at the corrected gestational age of 1 year, and a significantly lower abundance of Akkermansia one day before introducing solid food (P<0.05). CONCLUSIONS There are significant differences in the composition of intestinal microbiota between preterm infants with NDI and those without NDI. This study enriches the data on the characteristics of intestinal microbiota in preterm infants with NDI and provides reference for the microbiota therapy and intervention for NDI in preterm infants.
Infant ; Child ; Infant, Newborn ; Humans ; Child, Preschool ; Infant, Premature ; Prospective Studies ; Gastrointestinal Microbiome ; China ; Infant, Premature, Diseases ; Gestational Age

OBJECTIVE: Exposure to high intensity, low frequency noise (HI-LFN) causes vibroacoustic disease (VAD), with memory deficit as a primary non-auditory symptomatic effect of VAD. However, the underlying mechanism of the memory deficit is unknown. This study aimed to characterize potential mechanisms involving morphological changes of neurons and nerve fibers in the hippocampus, after exposure to HI-LFN. METHODS: Adult wild-type and transient receptor potential vanilloid subtype 4 knockout (TRPV4^-/-) mice were used for construction of the HI-LFN injury model. The new object recognition task and the Morris water maze test were used to measure the memory of these animals. Hemoxylin and eosin and immunofluorescence staining were used to examine morphological changes of the hippocampus after exposure to HI-LFN. RESULTS: The expression of TRPV4 was significantly upregulated in the hippocampus after HI-LFN exposure. Furthermore, memory deficits correlated with lower densities of neurons and neurofilament-positive nerve fibers in the cornu ammonis 1 (CA1) and dentate gyrus (DG) hippocampal areas in wild-type mice. However, TRPV4^-/- mice showed better performance in memory tests and more integrated neurofilament-positive nerve fibers in the CA1 and DG areas after HI-LFN exposure. CONCLUSION TRPV4 up-regulation induced neurofilament positive nerve fiber injury in the hippocampus, which was a possible mechanism for memory impairment and cognitive decline resulting from HI-LFN exposure. Together, these results identified a promising therapeutic target for treating cognitive dysfunction in VAD patients.
Animals ; Mice ; TRPV Cation Channels/metabolism* ; Intermediate Filaments/metabolism* ; Hippocampus/metabolism* ; Neurons/metabolism* ; Memory Disorders/metabolism*

OBJECTIVE: To investigate the effects of mTOR inhibitors everolimus (EVE) and gemcitabine (GEM) on the proliferation, apoptosis and cell cycle of diffuse large B-cell lymphoma (DLBCL) cell line U2932, and further explore the molecular mechanisms, so as to provide new ideas and experimental basis for the clinical treatment of DLBCL. METHODS: The effect of EVE and GEM on the proliferation of U2932 cells was detected by CCK-8 assay, the IC₅₀ of the two drugs was calculated, and the combination index (CI=) of the two drugs was calculated by CompuSyn software. The effect of EVE and GEM on apoptosis of U2932 cells was detected by flow cytometry with AnnexinV-FITC/PI staining. Flow cytometry with propidium iodide (PI) staining was used to detect the effect of EVE and GEM on the cell cycle of U2932 cells. Western blot assay was used to detect the effects of EVE and GEM on the channel proteins p-mTOR and p-4EBP1, the anti-apoptotic proteins MCL-1 and Survivin, and the cell cycle protein Cyclin D1. RESULTS: Both EVE and GEM could significantly inhitbit the proliferation of U2932 cells in a time- and dose-dependent manner (r=0.465, 0.848; 0.555, 0.796). According to the calculation of CompuSyn software, EVE combined with GEM inhibited the proliferation of U2932 cells at 24, 48 and 72 h with CI=<1, which had a synergistic effect. After treated U2932 cells with 10 nmol/L EVE, 250 nmol/L GEM alone and in combination for 48 h, both EVE and GEM induced apoptosis, and the difference was statistically significant compared with the control group (P<0.05). The apoptosis rate was significantly enhanced after EVE in combination with GEM compared with single-agent (P<0.05). Both EVE and GEM alone and in combination significantly increased the proportion of cells in G₁ phase compared with the control group (P<0.05). The proportion of cells in G₁ phase was significantly increased when the two drugs were combined (P<0.05). The expression of p-mTOR and effector protein p-4EBP1 was significantly downregulated in the EVE combined with GEM group, the expression of anti-apoptotic proteins MCL-1, Survivin and cell cycle protein cyclin D1 was downregulated too (P<0.05). CONCLUSION EVE combined with GEM can synergistically inhibit the proliferation of U2932 cells, and the mechanism may be that they can synergistically induce apoptosis by downregulating the expression of MCL-1 and Survivin proteins and block the cell cycle progression by downregulating the expression of Cyclin D1.
Humans ; Gemcitabine ; Everolimus/pharmacology* ; Survivin/pharmacology* ; Cyclin D1/pharmacology* ; Myeloid Cell Leukemia Sequence 1 Protein ; Cell Line, Tumor ; Cell Proliferation ; TOR Serine-Threonine Kinases ; Apoptosis ; Apoptosis Regulatory Proteins ; Cell Cycle Proteins ; Lymphoma, Large B-Cell, Diffuse