BACKGROUND: Peripheral helper T (T PH ) cells are uniquely positioned within pathologically inflamed non-lymphoid tissues to stimulate B-cell responses and antibody production. However, the phenotype, function, and clinical relevance of T PH cells in hepatocellular carcinoma (HCC) are currently unknown. METHODS: Blood, tumor, and peritumoral liver tissue samples from 39 HCC patients (Sep 2016-Aug 2017) and 101 HCC patients (Sep 2011-Dec 2012) at the Third Affiliated Hospital of Sun Yat-sen University were used. Flow cytometry was used to quantify the expression, phenotype, and function of T PH cells. Log-rank tests were performed to evaluate disease-free survival and overall survival in samples from 39 patients and 101 patients with HCC. T PH cells, CD19 + B cells, and T follicular helper (T FH ) cells were cultured separately in vitro or isolated from C57/B6L mice in vivo for functional assays. RESULTS: T PH cells highly infiltrated tumor tissues, which was correlated with tumor size, early recurrence, and shorter survival time. The tumor-infiltrated T PH cells showed a unique ICOS hi CXCL13 + IL-21 - MAF + BCL-6 - phenotype and triggered naïve B-cell differentiation into regulatory B cells. Triggering programmed cell death protein 1 (PD-1) induced the production of C-X-C motif chemokine ligand 13 (CXCL13) by T PH cells, which then suppressed tumor-specific immunity and promoted disease progression. CONCLUSION Our study reveals a novel regulatory mechanism of T PH cell-regulatory B-cell-mediated immunosuppression and provides an important perspective for determining the balance between the differentiation of protumorigenic T PH cells and that of antitumorigenic T FH cells in the HCC microenvironment.
Carcinoma, Hepatocellular/metabolism* ; Liver Neoplasms/metabolism* ; Humans ; T-Lymphocytes, Helper-Inducer/metabolism* ; Animals ; Mice ; Male ; Female ; Mice, Inbred C57BL ; Middle Aged ; B-Lymphocytes, Regulatory/metabolism* ; Flow Cytometry ; Interleukin-21 ; Aged ; Chemokine CXCL13/metabolism*

Gastric cancer (GC) is characterized by high morbidity and mortality rates. Chinese agarwood comprises the resin-containing wood of Aquilaria sinensis (Lour.) Gilg., traditionally utilized for treating asthma, cardiac ischemia, and tumors. However, comprehensive research regarding its anti-GC effects and underlying mechanisms remains limited. In this study, Chinese agarwood petroleum ether extract (CAPEE) demonstrated potent cytotoxicity against human GC cells, with half maximal inhibitory concentration (IC₅₀) values for AGS, HGC27, and MGC803 cells of 2.89, 2.46, and 2.37 μg·mL^-1, respectively, at 48 h. CAPEE significantly induced apoptosis in these GC cells, with B-cell lymphoma-2 (BCL-2) associated X protein (BAX)/BCL-2 antagonist killer 1 (BAK) likely mediating CAPEE-induced apoptosis. Furthermore, CAPEE induced G₀/G₁ phase cell cycle arrest in human GC cells via activation of the deoxyribonucleic acid (DNA) damage-p21-cyclin D1/cyclin-dependent kinase 4 (CDK4) signaling axis, and increased Fe²⁺, lipid peroxides and reactive oxygen species (ROS) levels, thereby inducing ferroptosis. Ribonucleic acid (RNA) sequencing, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting analyses revealed CAPEE-mediated upregulation of heme oxygenase-1 (HO-1) in human GC cells. RNA interference studies demonstrated that HO-1 knockdown reduced CAPEE sensitivity and inhibited CAPEE-induced ferroptosis in human GC cells. Additionally, CAPEE administration exhibited robust in vivo anti-GC activity without significant toxicity in nude mice while inhibiting tumor cell growth and promoting apoptosis in tumor tissues. These findings indicate that CAPEE suppresses human GC cell growth through upregulation of the DNA damage-p21-cyclin D1/CDK4 signaling axis and HO-1-mediated ferroptosis, suggesting its potential as a candidate drug for GC treatment.
Animals ; Humans ; Mice ; Antineoplastic Agents, Phytogenic ; Apoptosis/drug effects* ; Cell Line, Tumor ; Cyclin D1/genetics* ; Cyclin-Dependent Kinase 4/genetics* ; DNA Damage/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Ferroptosis/drug effects* ; G1 Phase Cell Cycle Checkpoints/drug effects* ; Heme Oxygenase-1/genetics* ; Mice, Inbred BALB C ; Mice, Nude ; Plant Extracts/pharmacology* ; Stomach Neoplasms/physiopathology* ; Thymelaeaceae/chemistry* ; Up-Regulation/drug effects*

ObjectiveThe antitumor activity of sesquiterpenoid M36 isolated from Myrrha against human hepatoma HepG2 cells was investigated in this study. MethodHepG2 cells were treated with M36 at different concentrations （0， 2， 4， 6， 8， 10 μmol·L^-1）. Firstly， the effects of M36 on the proliferation of human hepatoma HepG2 cells were detected by methyl thiazolyl tetrazolium （MTT）， colony formation assay， and EdU proliferation assay. Hoechst staining， flow cytometry analysis， and Western blot were used to explore the effect of M36 on the apoptosis of human hepatoma HepG2 cells. Acridine orange staining and western blotting were used to examine the effect of M36 on autophagy in HepG2 cells. Finally， Western blot was used to detect protein expression of cancer-related signaling pathways. ResultCompared with the blank group， M36 treatment significantly inhibited the proliferation of human hepatoma HepG2 cells （P<0.01）， and the half inhibitory concentration （IC₅₀） value of M36 for 48 h was 5.03 μmol·L^-1， in a dose- and time-dependent manner. M36 was also able to induce apoptosis and autophagy in human hepatoma HepG2 cells. After treatment with 8 μmol·L^-1 M36 for 48 hours， the apoptosis rate of HepG2 cells was （42.03±9.65）% （P<0.01）. Compared with the blank group， HepG2 cells treated with 4 and 8 μmol·L^-1 M36 for 48 h had a significant increase in cleaved poly ADP-ribose polymerase （cleaved-PARP） protein levels （P<0.01）. Acridine orange staining showed that autophagy was significantly activated in HepG2 cells treated with 4 and 8 μmol·L^-1 M36 for 48 h compared with the blank group （P<0.01）， which was further verified by the up-regulation of microtubule-associated protein 1 light chain 3 Ⅱ （LC3 Ⅱ）. Western blot results showed that compared with the blank group， the levels of phosphorylated extracellular regulated protein kinase （p-ERK）， phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）， phosphorylated c-Jun N-terminal kinase （p-JNK）， and its downstream nuclear transcription factors c-Jun and p-c-Jun protein were significantly increased in M36 group （P<0.05， P<0.01）. The mechanism may be related to the up-regulation of MAPK signaling pathway. ConclusionThe sesquiterpenoid M36 isolated from Myrrha inhibits the proliferation of human hepatoma HepG2 cells and promotes apoptosis and autophagy， which may be related to the activation of the MAPK signaling pathway.

OBJECTIVE To analyze the compositional differences between Fructus Tritici Levis and Triticum aestivum， and to provide reference for identification and quality control of both. METHODS Twenty batches of Fructus Tritici Levis and three batches of T. aestivum were collected， and their fingerprints were acquired by high-performance liquid chromatography and the similarities were evaluated by the Evaluation System of Similarity of Chromatographic Fingerprints of Traditional Chinese Medicine （2012 version）. Cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed to analyze the difference of Fructus Tritici Levis and T. aestivum from different regions， and the differential components were screened. The contents of the six identified components in Fructus Tritici Levis and T. aestivum were determined. RESULTS The similarities of the fingerprints of Fructus Tritici Levis ranged from 0.928 to 0.996， and the relative similarities of T. aestivum with Fructus Tritici Levis ranged from 0.761 to 0.773. A total of 19 common peaks were calibrated， and six components including linolenic acid， linoleic acid， 5-heptadecylresorcinol， 5-nonadodecylresorcinol， 5- heneicosylresorcinol， and 5-tricosylresorcinol were identified. The results of CA and PCA showed that Fructus Tritici Levis and T. aestivum could be clearly distinguished； the distribution of Fructus Tritici Levis from Anhui province was relatively concentrated. The results of OPLS-DA showed that linolenic acid， linoleic acid， and other six unknown compounds were the differential components between Fructus Tritici Levis and T. aestivum. The average contents of the six identified components in Fructus Tritici Levis were 0.100 9， 1.094 0， 0.005 1， 0.030 9， 0.098 2，and 0.024 8 mg/g， respectively； the contents of linolenic acid and linoleic acid in Fructus Tritici Levis were significantly higher than those in T. aestivum （P＜0.05）.CONCLUSIONS The established qualitative and quantitative methods are simple and reliable， and can be used for the identification and quality evaluation of Fructus Tritici Levis and T. aestivum. The identified differential components， such as linolenic acid and linoleic acid， can also provide clues for the differentiation and pharmacological study of Fructus Tritici Levis and T. aestivum.

OBJECTIVE To analyze the compositional differences between Fructus Tritici Levis and Triticum aestivum， and to provide reference for identification and quality control of both. METHODS Twenty batches of Fructus Tritici Levis and three batches of T. aestivum were collected， and their fingerprints were acquired by high-performance liquid chromatography and the similarities were evaluated by the Evaluation System of Similarity of Chromatographic Fingerprints of Traditional Chinese Medicine （2012 version）. Cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed to analyze the difference of Fructus Tritici Levis and T. aestivum from different regions， and the differential components were screened. The contents of the six identified components in Fructus Tritici Levis and T. aestivum were determined. RESULTS The similarities of the fingerprints of Fructus Tritici Levis ranged from 0.928 to 0.996， and the relative similarities of T. aestivum with Fructus Tritici Levis ranged from 0.761 to 0.773. A total of 19 common peaks were calibrated， and six components including linolenic acid， linoleic acid， 5-heptadecylresorcinol， 5-nonadodecylresorcinol， 5- heneicosylresorcinol， and 5-tricosylresorcinol were identified. The results of CA and PCA showed that Fructus Tritici Levis and T. aestivum could be clearly distinguished； the distribution of Fructus Tritici Levis from Anhui province was relatively concentrated. The results of OPLS-DA showed that linolenic acid， linoleic acid， and other six unknown compounds were the differential components between Fructus Tritici Levis and T. aestivum. The average contents of the six identified components in Fructus Tritici Levis were 0.100 9， 1.094 0， 0.005 1， 0.030 9， 0.098 2，and 0.024 8 mg/g， respectively； the contents of linolenic acid and linoleic acid in Fructus Tritici Levis were significantly higher than those in T. aestivum （P＜0.05）.CONCLUSIONS The established qualitative and quantitative methods are simple and reliable， and can be used for the identification and quality evaluation of Fructus Tritici Levis and T. aestivum. The identified differential components， such as linolenic acid and linoleic acid， can also provide clues for the differentiation and pharmacological study of Fructus Tritici Levis and T. aestivum.

Objective To analyze the medication law and academic thoughts of national TCM master Qian Ying in the treatment of chronic hepatitis B through data mining.Methods Totally 168 cases of chronic hepatitis B treated by Professor Qian Ying from Mar.2000 to Dec.2020 were retrospectively collected,and the properties,tastes and tropism meridians,core prescriptions and drug groups of the prescription drugs were analyzed by using the famous doctor inheritance platform.Results Totolly 168 medical cases involved 168 patients and 227 kinds of Chinese materia medica,with a total frequency of 2 158 times.The characteristics of properties,tastes and tropism meridians showed that the main property was cold,and the main taste was bitter,and the main meridian was liver meridian.34 kinds of high-frequency Chinese materia medica(mainly were tonics,heat-clearing medicines,and medicines for activating blood circulation and reducing stasis),28 kinds of core Chinese materia medica,10 pairs of highly co-occurring drugs,and 10 potential drug groups were mined.Conclusion Professor Qian Ying believes that the pathogenesis of chronic hepatitis B is deficiency in nature and excess in superficiality,and the treatment focuses on tonifying deficiency.It is often treated from the liver and emphasizes the harmonization of liver,spleen and kidney.Tonifying deficiency,clearing heat,promoting blood circulation and removing blood stasis are the main treatment methods,followed by dispelling dampness,promoting qi,eliminating phlegm,opening stagnation and relieving the exterior,etc.He pays attention to the harmonization of body and use,is good at using multiple methods.

Objective To explore the effect of Traditional Chinese Medicine's five-element music therapy com-bined with individualized emotional counseling intervention in patients with autoimmune liver disease of liver de-pression and spleen deficiency type.Methods 74 patients with autoimmune liver disease of liver depression and spleen deficiency type who were admitted to the hepatology department of a tertiary hospital from January 2021 to February 2022 were selected as research subjects,and they were divided into an experimental group and a control group by a random number table with 37 cases in each group.The control group was given routine treatment and nursing care,as well as Traditional Chinese Medicine's emotional nursing.The experimental group received Tradi-tional Chinese Medicine's five-element music therapy combined with individualized emotional counseling intervention on the basis of routine treatment and nursing care,with an intervention period of 4 weeks.The Generalized Anxiety Disorder Scale(GAD-7),Patient Health Questionnaire Depression Scale(PHQ-9)and Pittsburgh Sleep Quality Index(PSQI)were used to evaluate patients within 24 hours after admission and after 4 weeks of intervention.Results A total of 72 patients completed the study,with 36 in the experimental group and 36 in the control group.Before and after intervention,the scores of GAD-7,PHQ-9,and PSQI in the experimental group were compared,and the dif-ferences were statistically significant(P＜0.001).After the intervention,the GAD-7 score in the experimental group was(7.89±3.15),which was lower than(10.14±4.41)in the control group.The PHQ-9 score was(12.33±4.08)in the ex-perimental group,which was lower than(14.64±5.34)in the control group.The PSQI score was(12.39±3.06)points,which was lower than(14.22±3.37)in the control group,and the differences were statistically significant(P＜0.05).Conclusion The application of Traditional Chinese Medicine's five-element music therapy combined with individual-ized emotional guidance intervention can effectively improve the sleep quality and help alleviate negative emotions in patients with liver depression and spleen deficiency type of autoimmune liver disease.

Objective:To explore the role of information-based points system management platform in enhancing the medical management capabilities of laboratory medicine.Methods:A total of 106 personnel who were on duty from January 2021 to December 2022 in the clinical laboratory of Guangdong Provincial People′s Hospital were selected and divided into 8 professional groups, including 24 in Clinical Routine Lab, 22 in Clinical Biochemistry Lab, 11 in Clinical Immune Lab, 13 in Clinical Microbiology Lab, 10 in Clinical Molecular Lab, 9 in Flow Cytometry Lab, 9 in HuiFu Lab, and 8 in Interdisciplinary Lab. Information-based points system management platform was established by formulating the detailed rules for the assessment of this points system, and the points achieved by the 106 participants in 2021 and 2022 were obtained and analyzed. The points were divided into 6 intervals (<41, 41-45, 46-50, 51-55, 56-60 and>60).Results:The overall medical points of laboratory medicine was significantly higher in 2022 than in 2021 (52.15±5.48 vs 49.81±3.73, P<0.001). In 2022, the number of participants with points between 41-45 and 46-50 intervals was lower than those in 2021; with points 51-55, 56-60 and>60 intervals was higher than those in 2021 (χ2=10.479, P<0.05). In addition, the points of Clinical Molecular, Clinical Microbiology and HuiFu Lab were 57.46±7.40, 52.05±5.43 and 54.98±4.31 respectively, which were significantly higher than those of 2021 (50.16±4.14, 49.10±4.06, 49.23±4.28, respectively;all P<0.05). The number of report revisions has significantly decreased in 2022 (18) compared to that in 2021 (48), clinical communication and various levels of publicity and promotion were significantly increased from 52 and 58 (2021) to 108 and 137 (2022), respectively. Conclusion:The construction and application of the information-based points management system have effectively improved the enthusiasm and subjective initiative of laboratory personnel, enhanced the quality and service capabilities of laboratory medicine, and might serve as an important tool for standardized management of laboratory medicine.

Objective To study the medication law of Professor Qian Ying in the treatment of primary liver cancer based on data mining technology;To provide ideas for the clinical treatment of primary liver cancer.Methods Outpatient TCM prescriptions of Professor Qian Ying for the treatment of liver cancer from November 2008 to August 2020 were collected,and a data table was established after sorting.The drug frequency,property and taste and tropism were analyzed using Excel 2019.The medical case analysis module of the Great Physician Inheritance Platform was used to analyze the core drugs,the symbiosis analysis between drug pairs,the drug association analysis,and the drug clustering analysis of the screened TCM prescriptions.Results Totally 108 prescriptions were included,involving 188 kinds of Chinese materia medica,with a total frequency of 1 322 times.High-frequency drugs included Hedyotis Sinensis,Angelicae Sinensis Radix,Visci Herba,Curcumae Radix,Salviae Miltiorrhizae Radix et Rhizoma,etc.The medicinal properties were mainly cold,mild and warm,and the tastes were mainly bitter,sweet and pungent,and the main meridians were liver meridians,spleen meridians,kidney meridians and stomach meridians.There were 9 pairs of high frequency drug combinations in drug association,such as Curcumae Radix-Polygoni Orientalis Fructus,Visci Herba-Curcumae Rhizoma.In the correlation analysis of drug disease,the ones with higher correlations include"stomachache-Salviae Miltiorrhizae Radix et Rhizoma""abdominal mass-Paeoniae Radix Rubra and Citri Reticulatae Pericarpium""tinnitus-Adenophorae Radix,Lycii Fructus,Visci Herba""prolonged sublingual collaterals-Curcumae Rhizoma,Polygoni Orientalis Fructus,Salviae Miltiorrhizae Radix et Rhizoma"and so on.Drug clustering could be divided into three potential drug clusters.Conclusion Professor Qian Ying often uses heat-clearing drugs,tonifying drugs,and promoting qi and blood circulation drugs to treat liver cancer,with Huqi Powder as the main formula and modified according to the syndromes.Clearing heat and detoxifying,soothing liver and relieving depression,removing blood stasis and regulating collatrals are used to treat its symptoms,and tonifying qi and invigorating spleen,regulating liver and nourishing liver and kidney are used to treat its essence.

Abstract: Objective　To establish a risk prediction model for nosocomial infection in preterm very low birth weight infants, and conduct internal validation. Methods　A total of 206 cases of very low birth weight premature infants hospitalized in the Department of Neonatology of Union Hospital Affiliated to Tongji Medical College from January 2018 to June 2020 were included in this study, factors that may affect the nosocomial infection of children were collected, and the infants were divided into two groups according to whether there is nosocomial infection. The influencing factors were compared between the two groups, and multivariate Logistic regression analysis was performed after screening variables with LASSO regression. According to the results of multi factor analysis, the nomogram model was constructed and verified internally. Results　A total of 29 of 206 children had nosocomial infection (14.08%), and 33 pathogenic bacteria were detected, including 23 Gram-negative bacteria, 9 Gram-positive bacteria and 1 fungus. The results of multivariate logistic regression analysis based on LASSO regression showed that the risk factors for nosocomial infection of VLBW premature infants were 28-31+6 weeks of gestation, amniotic fluid pollution, mechanical ventilation, indwelling gastric tube, unreasonable use of antibiotics, and hospitalization time ≥ 7 days. The protective factors were Apgar score ≥ 7 points at 1 min and breast feeding accounting for 50% or more (P<0.05). The Area Under Curve (AUC) of ROC curve of nomogram model was 0.946 [95%CI(0.923, 1.000)]. The calibration curve showed that the probability of hospital infection predicted by the model was basically consistent with the actual probability. The decision curve showed that when the probability threshold of nomogram model to predict the risk of nosocomial infection of very low birth weight premature infants was 0-0.85, the net rate of return was greater than 0. Conclusion　Preterm infants with extremely low birth weight are at high risk of nosocomial infection, mainly affected by factors such as gestational weeks, hospitalization time, amniotic fluid pollution, etc. The nomogram model constructed by the above factors has high accuracy and discrimination for predicting nosocomial infection in such children.