Objective To investigate the effects of Aconite decoction （AD） on the viability and polarization of murine RAW264.7 macrophages induced by lipopolysaccharide （LPS） or interleukin-4 （IL-4）. Methods Cytotoxicity of AD was assessed by the CCK-8 assay. RAW264.7 cells were polarized toward M1 phenotype by LPS or M2 phenotype by IL-4, followed by treatment with varying concentrations of AD. Macrophage polarization was analyzed by flow cytometry. Quantitative PCR was performed to measure mRNA expression of polarization-associated markers （IL-6, iNOS, Arg1, and Ym1）. ELISA was used to quantify secreted cytokines （TNF-α and IL-10）in the supernatant. Results At non-toxic concentrations, IL-6 and iNOS mRNA levels in LPS-stimulated cells were significantly upregulated while Arg1 and Ym1 expression in IL-4-treated groups were downregulated by AD. Concurrently, TNF-α secretion in LPS-induced M1 polarization was enhanced but IL-10 production in IL-4-induced M2 polarization was suppressed by AD. Conclusion AD could promote macrophage proliferation and viability, augments LPS-driven M1 polarization, and inhibit IL-4-mediated M2 polarization, which provided experimental evidence for the potential application of AD in tumor immunotherapy.

OBJECTIVE To explore the mechanisms of the flavonoids from new "Zhe Eight Flavors" Quzhou Fructus Aurantii(PTFC) against hepatocellular carcinoma based on the prediction of network pharmacology and experimental verification. METHODS From TCMSP, TCMID, ETCM, BATMAN-TCM and SwissTargetPrediction databases, the potential target proteins of PTFC, including naringin, narirutin and neohesperidin were collected. Based on the GeneCards, CTD, Disgenet, and OMIM databases, a set of target proteins for hepatocellular carcinoma was constructed. Taking the intersection of potential target proteins of PTFC and target proteins of hepatocellular carcinoma, key target proteins were obtained and a protein-protein interaction network was established. Besides, GO function and KEGG pathway enrichment analysis on the core target proteins was performed and a Compounds-Targets-Pathways-Disease network was constructed. Through proliferation, cloning, wound healing, and migration experiments, the effects of PTFC on the viability of HepG2 liver cancer cells were analyzed. Using fluorescence probe staining the impacts of PTFC on the mitochondrial membrane potential and apoptosis of HepG2 were observed. Finally, the validation of the regulatory effect of PTFC on the key predicted target PRKCA were carried out through RT-qPCR. RESULTS Based on network pharmacology, a total of 217 potential target proteins for PTFC were screened, with 59 intersecting target proteins related to diseases, including ALB, ESR1, PRKCA, and others. GO functional and KEGG pathway enrichment analysis revealed that the PTFC target proteins were involved in 193 biological processes and 13 cancer-related signaling pathways. Experimental results demonstrated that PTFC could impact the proliferation, cloning, wound healing, and migration abilities of liver cancer cells, leading to a decrease in mitochondrial membrane potential and promoting cell apoptosis. The results of RT-qPCR confirmed a significant downregulation of PRKCA expression by PTFC, validating the predictions made by network pharmacology analysis. CONCLUSION This study has revealed the potential molecular mechanism of PTFC treating hepatocellular carcinoma via the PRKCA target, laying the foundation for clinical application of PTFC.

OBJECTIVE To study the inhibitory effect and mechanism of curcumin and its derivatives A and B on TGF-β induced LX-2 cell fibrosis. METHODS Established the liver fibrosis model of LX-2 cells induced by TGF-β(10 ng·mL−1).The effects on cell proliferation were detected by CCK-8. The effects on cell apoptosis was detected by flow cytometry. The effects on fibrosis related factors(Collagen I, Collagen Ⅳ, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, MMP2, MMP9, TIMP1 and TIMP2) protein expression and gene transcription levels were detected by Western blotting and q-PCR. RESULTS The curcumin and its derivative A and B had the inhibition effects on normal LX-2 cells, and the IC25 values were 15.7, 2.6, 10.2 μmol·L−1, respectively. Compared to the model group, the curcumin(15.7 μmol·L−1) and its derivative A(2.6 μmol·L−1) and B(10.2 μmol·L−1) had the significant inhibition effects on cell proliferation of the TGF-β induced LX-2 cells(P<0.05). The cell apoptosis rate of curcumin derivative B group was higher than the model group(P<0.05). Collagen I, Fibronectin, Vimentin, α-SMA, TGFβR1 and TIMP-1 protein expression levels in curcumin group were lower, while the protein expression level of MMP-9 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, TIMP-1 and TIMP-2 in curcumin derivative A group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, TIMP-1 and TIMP-2 in curcumin derivative B group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The gene transcription levels of Collagen I, Fibronectin, α-SMA and TIMP-1 in curcumin group were lower(P<0.05). The gene transcription levels of Collagen I, Fibronectin and α-SMA in curcumin derivative A and B groups were lower(P<0.05). CONCLUSION Curcumin and its derivatives A and B inhibit the abnormal activation and proliferation of TGF-β-induced LX-2 cells, inhibit the excessive secretion and accumulation of its extracellular matrix components, and promote its degradation, thus playing an anti-fibrotic effect in vitro, especially the curcumin derivative B.

OBJECTIVE: To evaluate the inhibitory effect of cordycepin on oral cancer xenograft in nude mice and explore the underlying mechanisms. METHODS: Sixteen BALB/c mice bearing subcutaneous human tongue squamous cell carcinoma (TSCC) TCA-8113 cell xenografts were randomized into model group and cordycepin treatment group for daily treatment with saline and cordycepin for 4 weeks. After the treatment, the tumor xenografts were dissected and weighed to assess the tumor inhibition rate. Histological changes in the heart, spleen, liver, kidney, and lung of the mice were evaluated with HE staining, and tumor cell apoptosis was examined using TUNEL staining; The expressions of Bax, Bcl-2, GRP78, CHOP, and caspase-12 in the xenografts were detected using RT-qPCR and Western blotting. RESULTS: Cordycepin treatment resulted in a tumor inhibition rate of 56.09% in the nude mouse models, induced obvious changes in tumor cell morphology and significantly enhanced apoptotic death of the tumor cells without causing pathological changes in the vital organs. Cordycepin treatment also significantly reduced Bcl-2 expression (P < 0.05) and increased Bax, GRP78, CHOP, and caspase-12 expressions at both the RNA and protein levels in the tumor tissues. CONCLUSION Cordycepin treatment can induce apoptotic death of TCA-8113 cell xenografts in nude mice via the endogenous mitochondrial pathway and endoplasmic reticulum stress pathways.
Humans ; Animals ; Mice ; Carcinoma, Squamous Cell/drug therapy* ; Heterografts ; Mice, Nude ; Tongue Neoplasms/drug therapy* ; Cordyceps ; Caspase 12 ; Endoplasmic Reticulum Chaperone BiP ; bcl-2-Associated X Protein ; Tongue

Objective:To establish and preliminarily validate a nomogram model for predicting the risk of retinal vein occlusion (RVO).Methods:A retrospective clinical study. A total of 162 patients with RVO (RVO group) diagnosed by ophthalmology examination in The Second Affiliated Hospital of Xi'an Jiaotong University from January 2017 to April 2022 and 162 patients with age-related cataract (nRVO group) were selected as the modeling set. A total of 45 patients with branch RVO, 45 patients with central RVO and 45 patients with age-related cataract admitted to Xi'an Fourth Hospital from January 2022 to February 2023 were used as the validation set. There was no significant difference in gender composition ratio ( χ2=2.433) and age ( Z=1.006) between RVO group and nRVO group ( P=0.120, 0.320). Age, gender, blood routine (white blood cell count, hemoglobin concentration, platelet count, neutrophil count, monocyte count, lymphocyte count, erythrocyte volume, mean platelet volume, platelet volume distribution width), and four items of thrombin (prothrombin time, activated partial thrombin time, fibrinogen, and thrombin time) were collected in detail ), uric acid, blood lipids (total cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein, lipoprotein a), hypertension, diabetes mellitus, coronary heart disease, and cerebral infarction. Neutrophil/lymphocyte ratio and platelet/lymphocyte ratio were calculated. The single logistic regression was used to analyze the clinical parameters of the two groups of patients in the modeling set, and the stepwise regression method was used to screen the variables, and the column graph for predicting the risk of RVO was constructed. The Bootstrap method was used to repeated sample 1 000 times for internal and external verification. The H-L goodness-of-fit test and receiver operating characteristic (ROC) curve were used to evaluate the calibration and discrimination of the nomogram model. Results:After univariate logistic regression and stepwise regression analysis, high density lipoprotein, neutrophil count and hypertension were included in the final prediction model to construct the nomogram. The χ2 values of the H-L goodness-of-fit test of the modeling set and the validation set were 0.711 and 4.230, respectively, and the P values were 0.701 and 0.121, respectively, indicating that the nomogram model had good prediction accuracy. The area under the ROC curve of the nomogram model for predicting the occurrence of post-stroke depression in the modeling set and the verification set was 0.741 [95% confidence interval ( CI) 0.688-0.795] and 0.741 (95% CI 0.646-0.836), suggesting that the nomogram model had a good discrimination. Conclusions:Low high density lipoprotein level, high neutrophil count and hypertension are independent risk factors for RVO. The nomogram model established based on the above risk factors can effectively assess and quantify the risk of post-stroke depression in patients with cerebral infarction.

Abstract OBJECTIVE To study the mechanism of NLRP3 gene’s regulation on inflammatory response in non-alcoholic fatty liver disease(NAFLD) mice induced by high-fat and high-fructose diet using NLRP3 gene knockout mice. METHODS Use male homozygous(NLRP3-/-) mice, and the high-fat and high-fructose diet was used to establish NAFLD model in NLRP3 knockout(KO) mice and wild-type(WT) mice, divided into KO high-fat and high-fructose diet(KO-HFD) group and WT high-fat and high-fructose diet(WT-HFD) group, while the WT and KO groups were also established. The body weight of mice in each group were observed. The changes of ALT, AST, TG, TC, MDA, SOD, lipidosis, apoptosis rate, IL-1β, IL-18, TNF-α, NF-κB, NLRP3, Caspase-1 and ASC in serum and tissue samples were tested to study the mechanism of the NLRP3 gene regulating the inflammatory response in NAFLD mice. RESULTS As time goes on, the mice weight of each group increased gradually, but the KO-HFD group increased less than the WT-HFD group. Each group’s level of ALT, AST, TG, TC in serum increased gradually, but the KO-HFD group increased less than the WT-HFD group. The level of MDA in liver tissues of each group was gradually increased and the level of SOD was gradually decreased, but the change range of KO-HFD group was smaller than that of WT-HFD group. The results of oil red O staining and Tunel section showed that the degree of lipid deposition and apoptosis in hepatocytes increased gradually in all groups, but the changes in KO-HFD group were less than that in WT-HFD group. The levels of serum and liver inflammatory factors IL-1β, IL-18, TNF-α and NF-κB increased in all groups, but the changes of KO-HFD group 20 weeks were less than those of WT-HFD group 20 weeks, and the difference was statistically significant. The expression levels of NLRP3 inflammasomes and related inflammatory cytokines NLRP3, Caspase-1, ASC, IL-1β and IL-18 in liver tissues of WT-HFD group were higher than those of KO-HFD group. The mRNA transcription levels of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in WT-HFD group were gradually increased, while there was no change in KO-HFD group. CONCLUSION The NLRP3 gene may be activated in NAFLD mice model, resulting in increased expression of NLRP3 inflammasome-associated protein, promoting the synthesis and secretion of downstream inflammatory factors, resulting in significant inflammatory response and liver damage, and promoting the progression of NAFLD disease.

Objective:To establish an animal model of chronic systemic inflammation with long-term high expression of circulating IL-6 by introducing exogenous IL-6 gene transfer vector.Methods:Recombinant murine IL-6-encoding adeno-associated virus (AAV-IL-6) was constructed. Twenty-one 24-week-old male C57BL/6J mice were randomly divided into three groups with seven in each group: AAV-IL-6 group, vector control (AAV-ctrl) group and blank control group. At 0, 8 and 16 weeks of intervention, the mice in the three groups were injected with AAV-IL-6 (100 μl 0.5×10 10 vp/ml), unloaded AAV (100 μl 0.5×10 10 vp/ml) and the same volume of saline in the tail vein, respectively. IL-6 levels in mouse serum were measured by ELISA. The general condition of mice was observed and blood routine tests were performed. Changes in blood biochemical parameters and C-reactive protein (CRP) levels were detected. At the end of 24-week intervention, the mice were sacrificed and the myocardium, liver, spleen, quadriceps femoris, knee joint and middle femur were taken for HE staining. Results:At 4, 8, 16 and 24 weeks after intervention, serum IL-6 levels were (75.41-169.28) pg/ml in the AAV-IL-6 group, while in the two control groups, the levels were below the lower limit of detection (7.8 pg/ml). At 24 weeks after intervention, the body weight of mice in the AAV-IL-6 group was significantly lower than that of mice in the two control groups; the neutrophil counts and CRP level in the AAV-IL-6 group were higher than those in the two control groups, while the levels of albumin, creatinine, triglyceride and cholesterol were lower than those in the two control groups. There were no differences in the aforementioned parameters between the two control groups. Compared with the blank control group, both AAV-IL-6 and AAV-ctrl groups showed increased lymphocyte counts. All mice had normal liver and kidney functions at the end of intervention. Histopathological findings indicated that the mice in the AAV-IL-6 group had focal infiltration of lymphocytes in the central venous area of the liver and around the myocardial and the skeletal muscle fibers, diffuse infiltration of multinucleated giant cells in the spleen, atrophic skeletal muscle, disorganized growth plate, reduced chondrocyte hypertrophic zone, thinner bone cortex and trabecular, and reduced osteoid. There were no histopathological changes in mice of the two control groups.Conclusions:Repeated tail vein injection of AAV-IL-6 could achieve long-term high expression of circulating IL-6 in mice, which manifested the phenotype of chronic systemic inflammation in preliminary detection and provided a safe, effective and simply accessible animal model for related studies.

Traumatic rib fractures are the most common injury in thoracic trauma. Previously，the patients with traumatic rib fractures were mostly treated non-surgically，of which 50%，especially those combined with flail chest presented chronic pain or chest wall deformities and over 30% had long-term disabilities，being unable to retain a full-time job. In the past two decades，thanks to the development of internal fixation material technology，the surgical treatment of rib fractures has achieved good outcomes. However，there are still some problems in clinical treatment，including inconsistency in surgical treatment and quality control in medical services. The current consensuses on the management of regional traumatic rib fractures published at home and abroad mainly focus on the guidance of the overall treatment decisions and plans，and relevant clinical guidelines abroad lacks progress in surgical treatment of rib fractures in recent years. Therefore，the Chinese Society of Traumatology affiliated to Chinese Medical Association and Chinese College of Trauma Surgeons affiliated to Chinese Medical Doctor Association，in conjunction with national multidisciplinary experts，formulate the Chinese Consensus for Surgical Treatment of Traumatic Rib Fractures（2021）following the principle of evidence-based medicine，scientific nature and practicality. This expert consensus puts forward some clear，applicable，and graded recommendations from aspects of preoperative imaging evaluation，surgical indications，timing of surgery，surgical methods，rib fracture sites for surgical fixation，internal fixation methods and material selections，treatment of combined injuries in rib fractures，in order to provide references for surgical treatment of traumatic rib fractures.

Objective: To determine the relationship between dengue virus load and clinical characteristics, so as to provide basis for dengue fever prevention and treatment. Methods : The dengue viral load and typing of 120 patients in Gongshu District of Hangzhou from June to November 2017 were detected by real-time fluorescent quantitative RT-PCR;the clinical indicators of these dengue patients were collected and their correlation with the viral load was analyzed. Results: The DNA detection of dengue virus in 120 patients showed that they were all typeⅡ. The median dengue virus load was 3.91×104 copies/mL. All the patients had fever, the average peak temperature was（38.96 ± 0.69）℃. There were 102（85.00%）cases with asthenia;116（96.67%）cases with white blood cell count（WBC）less than 4× 109/L;119（99.17%）cases with platelet count（PLT）less than 100×109/L;114（95.00%）cases with glutamic oxaloacetate transaminase（GOT）more than 40 U/L;81（67.50%）cases with glutamic pyruvate transaminase（GPT）more than 52 U/L;58（48.33%）cases with creatine kinase（CK）more than 210 U/L. There was no significant correlation of dengue virus load with length of hospitalization, peak temperature,duration of fever, WBC,PLT, GOT, GPT and CK（P>0.05）. There were 75（62.50%）severe patients, and their median viral load was 9.29×104copies/mL, which was higher than 5.33×103copies/mL in non-severe patients（P<0.05）. Conclusion The dengue virus load is not related with length of hospitalization,peak temperature,WBC,PLT,GOT,GPT and CK,but with the severity of the disease.

Objective Previous studies regarding the effects of tamoxifen ( TAM) on the thin endometrium are rare .The aim of this study was to explore the effects of TAM on patients with thin endometrium undergoing frozen thawed embryo transfer ( FET ) . Methods One hundred and thirty three patients with thin endometri-um undergoing FET treatment were recruited from January 2014 to June 2016, who canceled embryo transfer ( ET) or after FET due to thin endometrium in natural cycle or hormone replacement therapy cycle .Patients were randomly divided into letrozole ( LE,n=72) group or tamoxifen (TAM,n=61) group.All of the patients started to have oral pills of Estradiol Valerate 4 mg/d on the third day of menstruating cycles , then 6mg/d on the eighth day ,after 10~12 days then having ultrasonic monitoring of endometrial thickness and blood estradiol (E2), progesterone levels, It′s called endometrial preparation for hormone replacement cycle .To letrozole, tamoxifen group,the way of endometrial preparation were as follows:patients started to have oral pills of LE 2.5mg/d,TAM 40 mg/d on the third day of menstruating cycles for 5 days, then having ultrasonic monitoring and used drug of human chorionic gonadotropic hormone ,It′s called HCG day .After the dominant follicle ovulation then took progesterone intramuscular injection 40 mg/d, oral progesterone 20 mg/d to change endometrium ,then to transplant cleavage embryos or blastocysts after taking 3 or 5 days of progesterone , It′s called embryo transplanting day .The way of TAM endometrium preparation was called TAM cycle .The general data , hormone levels and clinical out-come between two groups were analyzed . Results The serum estradiol level of LE group both on HCG and transfer day [(1193.80± 629.64)ng/L vs (2776.30±157.34)ng/L;(1195.90±820.30)ng/L vs (2129.40±1208.71) ng/L,P=0.000] were statistically lower, serum luteinizing hormone level were statistically higher than TAM group [(20.48±15.50)IU/L vs (10.59±8.34)IU/L,P<0.05];im-plantation rate of LE group were statistically lower than TAM (39.32%vs 45.83%,P=0.001).The endometrial thickness and serum E 2 and P levels in TAM cycles were significantly higher compared with those in hormone replacement therapy cycle [(8.49±1.36)mm vs (6.43±0.96)mm,P=0.018]. Conclusion Tam compared LE with patients of thin endometrium undergoing FET can increased en -dometrial thickness and improve implantation rate ,thus Providing a new solution to thin endometrium .