Objective To analyze the characteristics and distribution of TCM syndromes of advanced gastric cancer;To provide reference for the standardization and clinical research of TCM syndromes of advanced gastric cancer.Methods The four diagnosis information with advanced gastric cancer was retrospectively collected at Dongzhimen Hospital of Beijing University of Chinese Medicine from January 2010 to December 2020.And the investigation results were analyzed by combining principal component analysis and clustering analysis,so as to explore the distribution pattern of TCM syndromes of advanced gastric cancer.Results Totally 164 patients were included,involving 601 case-times.10 principal components were obtained through principal component analysis on 29 items of four diagnosis information.The four diagnosis information with factor coefficient>0.4 were selected and allocated to the 10 principal components.Then,based on the results of principal component analysis,clustering analysis was conducted to obtain the distribution proportion of the three types of TCM syndromes.According to the syndrome differentiation by professional clinicians,the results were followed by the frequency distribution as cold coagulation and blood stasis(356,59.28%),stomach yin deficiency(145,24.17%)and phlegm-heat accumulation(100,16.55%).Conclusion There are three basic TCM syndromes of advanced gastric cancer,which are cold coagulation and blood stasis,stomach yin deficiency and phlegm-heat accumulation.Cold coagulation and blood stasis occupies the largest proportion,and the treatment should be based on warming the middle to dissipate cold and promoting blood circulation to remove blood stasis.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Background::Liver cancer remains the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths worldwide, causing a heavy burden globally. An updated assessment of the global epidemiology of the liver cancer burden that addresses geographical disparities is necessary to better understand and promote healthcare delivery.Methods::Data were extracted from the GLOBOCAN 2022 database, including the number, crude, and age-standardized rates of incidence and mortality at the global, country, continent, and human development index (HDI) regional levels. Age-standardized rates (incidence and mortality) per 100,000 person-years were adjusted based on the Segi-Doll World standard population. The mortality-to-incidence ratios (MIR) for each region and country were calculated. The HDI and gross national income (GNI) for 2022 were obtained, and a Pearson correlation analysis was conducted with the incidence, mortality, and MIR.Results::In 2022, approximately 866,136 new liver cancer cases and 758,725 related deaths were recorded worldwide, with a global MIR of 0.86. Males had a disproportionately higher burden than females across all levels, and the highest burden was observed in the elderly population. Geographically, the regions with the highest incidence rates included Micronesia, Eastern Asia, and Northern Africa, and the regions with the highest mortality rates included Northern Africa, Southeastern Asia, Eastern Asia, and Micronesia. Notably, Mongolia had a strikingly high burden compared to other countries. The highest MIR was observed in North America and the lowest in Africa. Negative associations of HDI and GNI with liver cancer mortality and MIR were identified, irrespective of sex.Conclusions::The current liver cancer burden underscores the presence of remarkable geographic heterogeneity, which is particularly evident across countries with varying HDI levels, highlighting the urgent need to prioritize health accessibility and availability to achieve health inequities.

"Toxicity Testing in the 21st Century—A Vision and Strategy"proposed by the National Research Council of US has brought innovative directives and objectives for toxicity evaluation and risk assessment,pushing forward the next generation of toxicity testing and risk assessment.In this initiative,the concept of adverse outcome pathways(AOPs)has emerged as a prominent methodology,capturing the attention of toxicologists and researchers due to its promising applications in recent years.The quantitative AOP(qAOP)is an extension of the adverse outcome pathway,which is built upon the foundational qualitative adverse outcome pathway model and leverages mathematical frame-works to depict dose-response and/or response-response relationships.This article reviews the princi-ples and advancement surrounding qAOP,introduceds two prevalent methodologies for constructing qAOP,Bayesian network models and regression models,and demonstrates diverse applications of qAOP.Actual cases are used to underscore the transformative role of qAOP in contemporary toxicology and risk assessment practices.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Objective:To investigate the role of group 3 innate lymphoid cells (ILC3) in skin wound healing, and to explore the underlying mechanisms.Methods:Twenty-four 5-week-old male C57BL/6 mice were randomly and equally allocated into 3 groups: the skin wound + ILC3 inhibitor group (referred to as ILC3 inhibitor group), the skin wound group, and the control group, with 8 mice in each group. Four days before the establishment of the wound model, mice in the ILC3 inhibitor group were intraperitoneally injected with 1 μg of ILC3 inhibitor every 2 days for a total of 2 doses, mice in the skin wound group were injected with an equal volume of physiological saline solution, and mice in the control group were fed normally. To establish a mouse skin wound model, a full-thickness circular incision with a diameter of 0.6 cm was made around the midpoint of the dorsal midline using a biopsy punch after the intraperitoneal injection of anesthetics, which was histologically confirmed to be a full-thickness injury. The size of the wounds was observed and recorded, photographs of the wounds were taken on days 0, 1, 3, 5, 7, and 9 after wounding, and corresponding wound healing rates were calculated. On day 9 after wounding, tissue samples were collected from the wound edges, and subjected to flow cytometry analysis to quantify ILC3 infiltrating around the skin wound, and hematoxylin and eosin (HE) staining was performed to assess the healing status of the skin wounds. Real-time quantitative polymerase chain reaction (qRT-PCR) was conducted to determine the mRNA expression of vitamin D receptor (VDR), Notch1, tumor necrosis factor-alpha (TNF-α), interleukin (IL) -17A, IL-17F, and IL-22 in the wound-edge tissues, and Western blot analysis to determine their protein expression. Statistical analysis was carried out by using one-way analysis of variance and t test. Results:On day 9 after wounding, the skin wound group showed an increased number of ILC3 in the wound-edge tissues (5.31% ± 1.47% vs. 3.10% ± 0.54%, P < 0.01), increased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F (all P < 0.05), but decreased mRNA and protein expression of VDR (both P < 0.05) compared with the control group; the protein expression of Notch1 was significantly higher in the skin wound group than in the control group ( P < 0.05), but there was no significant difference in its mRNA expression between the two groups ( P > 0.05). On days 1, 3 and 5, the wound healing rates were significantly higher in the ILC3 inhibitor group (45.17% ± 9.90%, 61.58% ± 11.61%, 75.61% ± 9.12%, respectively) than in the skin wound group (25.87% ± 10.96%, 47.78% ± 13.81%, 64.55% ± 10.29%, respectively, all P < 0.05). On day 9, the ILC3 inhibitor group showed a decreased number of ILC3 around the wound (2.69% ± 0.95%, P < 0.01), decreased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F in the wound-edge tissues (all P < 0.05), but increased mRNA and protein expression of Notch1 and VDR in the wound-edge tissues (all P < 0.05) compared with the skin wound group. On day 9 after wounding, histopathological examination with HE staining revealed continuous and intact epithelial structure, as well as dense and neatly arranged collagen fibers in the ILC3 inhibitor group, and the structures of hair follicles, blood vessels, and sebaceous glands were similar to those in the control group. Conclusions:Skin ILC3 infiltrated local wounds and were involved in the skin wound healing process through inflammatory factors such as TNF-α, IL-17A, IL-17F, and IL-22. Downregulating the number of ILC3 may promote skin wound healing by activating VDR and Notch1, as well as inhibiting the TNF-α signaling pathway and the expression of downstream inflammatory factors.

Objective:To investigate the change trend of etiological burden of disease of liver cancer in the Chinese population from 1990 to 2019.Methods:The descriptive epidemiologic method was conducted. Based on the Global Burden of Disease data from the Institute for Health Metrics and Evaluation at the University of Washington, the data related to liver cancer burden caused by hepatitis B virus (HBV) infection, hepatitis C virus (HCV) infection, alcohol, nonalcoholic steatohepatitis (NASH) and other factors, including number of new cases, the crude incidence rate, age-specific incidence rate, number of deaths, crude mortality rate and age-specific mortality rate, in the Chinese population from 1990 to 2019 were collected. The age-standardized rate was calculated based on the world standardized population structure in 2019 from the Global Burden of Disease data. Observation indicators: (1) the incidence of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (2) the mortality of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (3) the change trend of age-specific incidence rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (4) the age-specific mortality rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. Count data were expressed as absolute numbers, percentages and ratio. Based on the junction point regression model, the Joinpoint software (V.4.9.1.0) was used to calculate the annual percentage change, average annual percentage change (AAPC) and 95% confidence intervals ( CI) of age-specific incidence rate and age-specific mortality rate of liver cancer caused by different etiologies. Results:(1) The incidence of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the number of new cases of liver cancer in Chinese population decreased from 236 825 to 210 462, and the crude incidence rate decreased from 20.01/100,000 to 14.80/100,000. The new cases of liver cancer caused by HBV infection, HCV infection and other factors showed a downward trend, and the absolute change rates were ?14.76%, ?3.98% and ?26.67%, respectively. The new cases of liver cancer caused by alcohol and NASH showed a increase trend, and the absolute change rates were 9.31% and 13.91%, respectively. (2) The mortality of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the number of deaths of liver cancer in Chinese population decreased from 232 449 to 187 700, and the crude mortality rate decreased from 19.64/100,000 to 13.20/100,000. The number of deaths of liver cancer caused by HBV infection, HCV infection and other factors showed a down-ward trend, and the absolute change rates were ?23.34%, ?10.99% and ?33.75%, respectively. The number of deaths of liver cancer caused by alcohol showed a slow downward trend, and the absolute change rate was ?0.51%. The number of deaths of liver cancer caused by NASH showed a increase trend, and the absolute change rate was 6.03%. (3) The change trend of age-specific incidence rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the AAPC of age-specific incidence rate of liver cancer caused by HBV infection, HCV infection, alcohol, NASH and other factors was ?3.61%(95% CI as ?4.10% to ?3.11%), ?3.57%(95% CI as ?3.99% to ?3.14%), ?2.79%(95% CI as ?3.24% to ?2.33%), ?2.65%(95% CI as ?3.09% to ?2.21%) and ?3.62%(95% CI as ?4.05% to ?3.19%), respectively. (4) The age-specific mortality rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the AAPC of age-specific mortality rate of liver cancer caused by HBV infection, HCV infection, alcohol, NASH and other factors was ?3.92%(95% CI as ?4.42% to ?3.41%), ?3.90%(95% CI as ?4.45% to ?3.35%), ?3.15%(95% CI as ?3.71% to ?2.58%), ?2.86%(95% CI as ?3.34% to ?2.38%) and ?4.09%(95% CI as ?4.64% to ?3.55%), respectively. Conclusions:From 1990 to 2019, the liver cancer burden of the Chinese population shows an overall downward trend, in which the liver cancer burden caused by HBV and HCV infection decreases the most, but HBV and HCV infection is still the main reason for the heavy burden of liver cancer. The age-specific incidence rate and age-specific mortality rate of liver cancer caused by alcohol and NASH show a downward trend, but the number of new cases of liver cancer caused by alcohol and NASH shows significant growth. The liver cancer burden caused by other factors shows a downward trend.

Stem cell research has become a frontier in the field of life sciences, and provides an ideal model for exploring developmental biology problems such as embryogenesis, histiocytosis, and gene expression regulation, as well as opens up new doors for clinical tissue defective and inheritance diseases. Among them, menstrual blood-derived stem cells (MenSCs) are characterized by wide source, multi-directional differentiation potential, low immune rejection characteristics. Thus, MenSCs can achieve individual treatment and have the most advantage of the clinical application. The central nervous system, including brain and spinal cord, is susceptible to injury. And lethality and morbidity of them tops the list of all types of trauma. Compared to peripheral nervous system, recovery of central nervous system after damage remains extremely hard. However, the treatment of stem cells, especially MenSCs, is expected to solve this problem. Therefore, biological characteristics of MenSCs and their treatment in the respect of central nervous system diseases have been reviewed at home and abroad in recent years, so as to provide reference for the treatment of central nervous system diseases.

Objective To investigate the mechanism of cellular senescence in ischemia reperfusion injury (IRI)-induced acute kidney injury (AKI) that leads to chronic kidney disease (CKD) in elderly mice.Methods An acute kidney injury model was established in C57B1/6 male mice at ages 8-10 weeks (young group) or 20-24 months (old group) by bilateral IRI.The animals were randomly divided into 4 groups as follows:Young-Sham (n=8),Old-Sham (n=8),Young-IRI (n=8),and Old-IRI groups (n=8).All mice were weighted,and their blood was collected from the tail vein at days 1,3,and 7 after surgery.The mice were killed on day 14 after surgery,and their kidneys were harvested for further analysis.Serum was used for the creatinine test.The changes of the renal tissue morphology and pathology were assessed using hematoxylin-eosin staining and sirius red staining.Immunofluorescence staining of collagen Ⅰ,F4/80,phosphor-histone H3 (p-HH3),and Ki67 were performed to determine the stage of the collagen deposit,macrophage filtration,and cell cycle G2/M arrest.The collagen Ⅰ expression was analyzed using western blot.The expression levels of TNF-α,IL-6,TGF-β,and collagen Ⅰ were determined using real-time PCR.Results Compared with that in the sham group,the serum creatinine levels in both Young-IRI and Old-IR1 groups were obviously increased.The Young-IRI group recovered completely on day 7.The Old-IRI group had higher creatinine levels than the Young-IRI group at each time point.Morphology and pathology analyses revealed that acute injury was repaired in the Young-IRI group,but slight inflammatory cell filtration and collagen deposition were observed in the Old-Sham and Old-IRI groups,respectively.Immunofluorescence staining revealed some F4/80-positive macrophage filtration,collagen Ⅰ deposition,and p-HH3 and Ki67 double-positive nuclear tubular epithelial cells in the Old-Sham group,but considerably more positive results were found in the Old-IRI group.Western blot analysis revealed that collagen Ⅰ expression level was higher in the Old-IRI group than in the Young-IRI group (P ＜ 0.01) and in the Old-Sham group than in the Young-Sham group (P ＜ 0.05).Real-time PCR demonstrated that the mRNA expression levels of cytokines and fibrosis markers,including of TNF-α,IL-6,TGF-β,and collagen Ⅰ,in the Old-Sham and Old-IRI groups were increased as compared with those in the Young-Sham and Young-IRI groups (P ＜ 0.05).Conclusions The levels of kidney inflammation,fibrosis,and cell-cycle arrest are lower in the old mice.After IRI injury,a sustained and ongoing inflammatory reaction is involved and more cells are arrested in the cell cycle G2/M,which inhibit renal repair and promote fibrosis progression.

AIM To establish an HPLC-DAD method for the simultaneous content determination of four constituents in Compound Kendir Leaves Tablets Ⅰ (Apocyni veneti Folium,Chrysanthemi indici Flos,Stephaniae tetrandrae Radix,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu VP-ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of methanol-0.5％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 260 nm and 325 nm.RESULTS Chlorogenic acid,hydrochlorothiazide,buddleodide and promethazine hydrochloride showed good linear relationships within the ranges of 24.91-498.2 ng (r =0.999 9),286.33-5 726.7 ng (r =0.999 9),10.04-200.9 ng (r =0.999 9) and 154.80-3 096.1 ng (r =0.999 9),whose average recoveries were 98.3％ (RSD =1.3％),99.1％ (RSD =0.6％),98.5％ (RSD =1.0％) and 99.3％ (RSD =1.2％),respectively.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Compound Kendir Leaves Tablets Ⅰ.