Objective　To investigate the role of granulocyte-macrophage colony-stimulating factor (GM-CSF) deficiency in the pathogenesis of Exophiala oligosperma (E. oligosperma) infection, a dematiaceous fungus, aiming to provide new insights and evidence for the treatment of dematiaceous fungal infections. Methods　C57BL/6 wild-type (WT) mice and Csf2 gene knockout (KO) mice (C57BL/6 background) were selected. Using E. oligosperma isolated from patients with caspase recruitment domain-containing protein 9 (CARD9) gene deficiency, a murine subcutaneous infection model was established to simulate the human infection route. The natural progression of infection in the mice was observed for six weeks, with skin lesion tissues collected at appropriate time points for pathological analysis and monitoring immune responses. Results　Both WT and Csf2 KO mice exhibited spontaneous pathogen clearance and gradual recovery of foot tissue appearance during the progression of infection, with a 100% survival rate at the end of observation. Compared to WT mice, Csf2 KO mice showed reduced footpad swelling at 1 and 2 weeks post-infection (t=4.674, t=5.961, P<0.01). Fungal clearance in Csf2 KO mice was delayed, with fungal colonies still detectable in lesion tissues at week 4, and Periodic Acid-Schiff (PAS)-positive fungal spores observed in histopathological sections. There was no significant difference found in macrophage infiltration between WT and Csf2 KO mice during the early stages of infection (1-2 weeks) (P>0.05), while neutrophil infiltration was significantly reduced in Csf2 KO mice at week 2 (t=3.287, P<0.01). In addition, Csf2 KO mice exhibited lower levels of IL-6 and IL-1β in foot lesion homogenates at week 1 (t=4.686, t=4.102, P<0.05). Conclusions　This study demonstrated that GM-CSF deficiency delays pathogenic fungi clearance, prolongs the disease course and affects early inflammatory cytokine production as well as neutrophil infiltration during the early stages of fungal infections.

Objective:To evaluate an effective manner to replace the drainage tube due to drainage complications during the irrigation treatment of abdominal infection after pancreaticoduodenectomy (PD).Methods:Clinical data of 39 patients with abdominal infection after PD due to drainage complications who were successfully treated by replacement of the drainage tubes with continued flushing in the Department of Hepatobiliary Surgery, Yuncheng Central Hospital Affiliated to Shanxi Medical University from August 2020 to August 2023 were retrospectively analyzed, including 23 males and 16 females, aged (54.8±9.6) years. According to the fashion of tube replacement, patients were divided into the observation group ( n=21), in which a flushing tube was placed through the original abdominal drainage tube sinus tract combined with external negative pressure suction; and the control group ( n=18), in which two parallel drainage tubes were placed through the original abdominal drainage tube sinus tract for flushing and drainage. The two groups were compared in terms of indicators before tube replacement, including the primary tumor classification, incidence of pancreatic fistula and biliary fistula after PD, complications of abdominal drainage, time from the surgery to tube replacement; and indicators after tube replacement, including total hospital stay, hospitalization cost, continuous abdominal lavage time, fever and elevated white blood cell count, number of dressing changes, etc. Results:There were no significant difference in the primary tumor classification, incidence of pancreatic fistula and biliary fistula after PD, complications of abdominal drainage, and time from PD to tube replacement between the two groups before tube replacement (all P>0.05). After tube replacement, the total hospitalization time(32.7±1.9 vs 44.7±14.5, d), hospitalization cost (67 604±16 052 vs 91 845±19 826, yuan), continuous abdominal lavage time [4.0 (3.0, 5.0) vs 9.0 (8.0, 10.8), d], fever [23.8% (5/21) vs 55.6% (10/18)] and leukocytosis rate [28.6% (6/21) vs 66.7% (12/18)], and times of dressing change times [10.0 (7.0, 13.0) vs 22.0 (18.2, 27.0)] of the observation group were lower than those of the control group (all P<0.05). Conclusion:Inserting a flushing tube through the sinus tract of original abdominal drainage tube combined with external negative pressure drainage is an effective way to manage the abdominal infection after PD surgery due to drainage complications, featuring good irrigation and drainage, short irrigation time, and early control of the abdominal infection.

Objective:To explore the effects of adriamycin（Adc）combined with hyperbaric oxygen（HBO）on the apoptosis of HepG2 cells and immunomodulatory factors in transplanted hepatocellular carcinoma（HCC）tissues in nude mice.Methods:The model of transplanted HCC was established in 20 nude mice，then the mice were divided into four groups：control group，HBO group，Adc group，and HBO + Adc group，with five mice in each group. The control group was reared normally. The HBO group was treated with HBO at 0.20 MPa for 1 h；the Adc group was treated with 10 mg/kg of Adc by gavage；the HBO + Adc group was treated with HBO at 0.20 MPa for 1 h and then given 10 mg/kg of Adc by gavage. All groups were treated once every other day for 21 days. After drug withdrawal，the tumors were dissected from the sacrificed mice，of which the tumor volume and weight were measured. The apoptosis of HepG2 cells was detected by TUNEL. The changes of apoptosis-related proteins in HepG2 cells were detected by western blotting. The changes of the contents of immunomodulatory factors in transplanted tumor tissues of nude mice were detected by ELISA.Results:After 21 days，the tumor volume and weight in the HBO + Adc group were lower than those in the other three groups（ P<0.05）. The proportion of apoptotic cells in the transplanted tumor tissues of nude mice of the HBO + Adc group was the highest. In the HBO + Adc group，the protein expression levels of Bax，caspase-3，and Cytochrome C were significantly increased；the protein expression of Bcl-2 was significantly reduced（ P<0.01）；and the IL-6 was significantly reduced；while the IL-18 and IFN-γ were significantly increased（ P<0.05）. Conclusion:The Adc combined with HBO can promote apoptosis of HepG2 cells in transplanted HCC tissues in nude mice，and effectively regulate the immunomodulatory factors to promote the recovery of the body’s immune function，suggesting obvious sensitizing and detoxifying effects.

Objective:To explore the effects of adriamycin（Adc）combined with hyperbaric oxygen（HBO）on the apoptosis of HepG2 cells and immunomodulatory factors in transplanted hepatocellular carcinoma（HCC）tissues in nude mice.Methods:The model of transplanted HCC was established in 20 nude mice，then the mice were divided into four groups：control group，HBO group，Adc group，and HBO + Adc group，with five mice in each group. The control group was reared normally. The HBO group was treated with HBO at 0.20 MPa for 1 h；the Adc group was treated with 10 mg/kg of Adc by gavage；the HBO + Adc group was treated with HBO at 0.20 MPa for 1 h and then given 10 mg/kg of Adc by gavage. All groups were treated once every other day for 21 days. After drug withdrawal，the tumors were dissected from the sacrificed mice，of which the tumor volume and weight were measured. The apoptosis of HepG2 cells was detected by TUNEL. The changes of apoptosis-related proteins in HepG2 cells were detected by western blotting. The changes of the contents of immunomodulatory factors in transplanted tumor tissues of nude mice were detected by ELISA.Results:After 21 days，the tumor volume and weight in the HBO + Adc group were lower than those in the other three groups（ P<0.05）. The proportion of apoptotic cells in the transplanted tumor tissues of nude mice of the HBO + Adc group was the highest. In the HBO + Adc group，the protein expression levels of Bax，caspase-3，and Cytochrome C were significantly increased；the protein expression of Bcl-2 was significantly reduced（ P<0.01）；and the IL-6 was significantly reduced；while the IL-18 and IFN-γ were significantly increased（ P<0.05）. Conclusion:The Adc combined with HBO can promote apoptosis of HepG2 cells in transplanted HCC tissues in nude mice，and effectively regulate the immunomodulatory factors to promote the recovery of the body’s immune function，suggesting obvious sensitizing and detoxifying effects.

Objective: To evaluate the feasibility and efficacy of naked-eye assessment (NA) of ultrasound-guided fine needle aspiration cytology(US-FNAC) smears, which was performed by a trained non-cytological physician. Methods: A total of 290 smears of FNAC in 143 thyroid nodules were used to evaluate the value of NA by an assistant with more than two years experience of intervention with ultrasound guidance. NA results such as the background of smear (bloody/non-bloody), thickness (thick/thin), as well as the contents (granulated/non-granulated) were recorded. The correlation between NA and cytological results was analyzed. Number of cells under microscopy, the non-diagnostic rate, and the significance between benignity and malignancy with different features of specimens were compared. Results: There was no significant difference between the NA background and cytological findings(P=0.707, 0.200, 0.705, respectively). There was significance in number of cells, non-dianostic rate between thick and thinner specimens (P=0.000, 0.001, respectively). However, there was no significance in benignity and malignancy between thick and thinner specimens (P=0.385). There was significance in cytology results between the granulated and non-granulated smears(all P=0.000). The difference was found in 51.1% of the smears with granulates were malignancy in cytology. While 77.8% of the non-granulated smears were ultimately diagnosed as benignity. Conclusions The NA could be performed by a trained non-cytological physician and contribute to preliminary predictions of cytology effect.

Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.

Objective:To explore the relationship between YKL-40 and the proliferation of breast cancer and its mechanism.Methods:The expression of YKL-40 in breast cancer MCF-7 cells was detected by immunofluorescence assay.Fluorescence microscope was used to observe the conversion efficiency,and Real-time PCR was used to screen the most effective YKL-40 siRNA.Expression levels of PI3K,P-PI3K,AKT,P-AKT of the PI3K/AKT pathway associated proteins was test by Western blot.At the same time,MTT and flow cytometry were validated by YKL-40 siRNA treatment of human breast cancer MCF-7 ceils,the differences of 24h,48h and 72h groups of cell proliferation ability and cell cycle.Result:MCF-7 cell express YKL-40 protein,mainly located in the cytoplasm.Real-time PCR show that siRNA01,siRNA02,siRNA03 compared with NC group YKL-40 gene silencing effect is remarkable.Among of them the strongest silencing effect is siRNA02 (P＜0.01),Western blot show the experimental group than the control group,Total PI3K and AKT remain unchanged while P-PI3K,P-AKT expression decreased (P＜0.05).In the experimental group,the number of G1 cells in the control group was increased (P＜0.01),while the S phase cells decreased (P＜0.01).MTT results showed that the experimental group compared with the control group,the proliferation ability is decreased(P＜0.01).Conclusions This study suggests that YKL-40 can be used as the upstream regulatory factor of PI3K/AKT signaling pathway and affect the process of cell cycle in breast cancer,and then regulate the proliferation of breast cancer,YKL-40 may be a crucial target for the treatment of breast cancer.

Sorafenib is a molecular targeted drug for the treatment of advanced primary liver cancer.However,along with the occurrence of drug resistance,the therapeutic effect was effected.At present,there is clear evidence that the emergence of drug resistance of live cancer is closely related to the epithelial-mesenchymal transition,liver cancer stem cells and the heterogeneity of liver cancer,and the PI3K/AKt signaling pathway as the vital common signaling channel was involved in the above mentioned process.From this we can conclude that complementary inhibtion of PI3K/AKt signaling pathway at the same time is the method that can strengthen the effect of sorafenib on the treatment of liver cancer so far.