ObjectiveTo explore the material basis of the "interaction of blood stasis and toxin" mechanism in cerebral ischemia-reperfusion injury， as well as the protective role of Huoxue Huayu Jiedu prescription （HXHYJDF） against ferroptosis. MethodsSixty SPF-grade male SD rats were randomly divided into six groups： sham group， model group， deferoxamine （DFO） group （100 mg·kg^-1）， low-dose HXHYJDF group （4.52 g·kg^-1）， medium-dose HXHYJDF group （9.04 g·kg^-1）， and high-dose HXHYJDF group （18.07 g·kg^-1）， with ten rats in each group. Except for the sham group， the other groups were used to replicate the model of focal cerebral ischemia-reperfusion in the middle cerebral artery of rats by the reforming Longa method. Neurological function was assessed at 1^st， 3^rd， 5^th， and 7^th days post-reperfusion using the modified neurological severity scores （m-NSS）. Brain tissue pathology and the morphology of mitochondria were observed using hematoxylin-eosin （HE） staining and transmission electron microscopy. The contents of malondialdehyde （MDA）， glutathione （GSH）， divalent iron ions （Fe²⁺）， and reactive oxygen species （ROS） in the ischemic cerebral tissue were detected using enzyme-linked immunosorbent assay （ELISA）. Immunohistochemistry and Western blot （WB） were used to detect the expression of iron death marker proteins glutathione peroxidase 4 （GPX4）， ferroportin-1 （FPN1）， transferrin receptor protein 1 （TfR1）， and ferritin mitochondrial （FtMt） in brain tissue. ResultsCompared with the sham group， the mNSS score of the model group was significantly increased （P<0.01）. HE staining showed that the number of neurons in the cortex of brain tissue was seriously reduced， and the intercellular space was widened. The nucleus was fragmented， and the cytoplasm was vacuolated. The results of transmission electron microscopy showed that the mitochondria in the cytoplasm contracted and rounded， and the mitochondrial cristae decreased. The matrix was lost and vacuolated， and the density of the mitochondrial bilayer membrane increased. The results of ELISA showed that the content of GSH decreased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS increased significantly （P<0.01）. The results of immunohistochemistry and WB showed that the expression of GPX4 and FPN1 proteins was significantly decreased （P<0.01）， and the expression of FtMt and TfR1 proteins was significantly increased （P<0.01）. Compared with those of the model group， the m-NSS scores of the high-dose and medium-dose HXHYJDF groups began to decrease on the 3^rd and 5^th days， respectively （P<0.05， P<0.01）. The results of HE and transmission electron microscopy showed that the intervention of HXHYJDF improved the pathological changes of neurons and mitochondria. The results of ELISA showed that the content of GSH in the medium-dose and high-dose HXHYJDF groups increased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS decreased significantly （P<0.05， P<0.01）. The content of GSH in the low-dose HXHYJDF group increased significantly （P<0.01）， and the contents of MDA and ROS decreased significantly （P<0.01）. The results of immunohistochemistry showed that the expression of GPX4 and FPN1 in the high-dose HXHYJDF group increased significantly （P<0.01）， and the expression of FtMt and TfR1 decreased significantly （P<0.01）. The expression of GPX4 and FPN1 in the medium-dose HXHYJDF group increased significantly （P<0.05）， and the expression of TfR1 decreased significantly （P<0.01）. WB results showed that the expression levels of FPN1 and GPX4 proteins in the high-dose， medium-dose， and low-dose HXHYJDF groups were significantly up-regulated （P<0.01）， and the expression levels of FtMt and TfR1 proteins were significantly down-regulated （P<0.01）. ConclusionHXHYJDF can significantly improve neurological dysfunction symptoms in rats with cerebral ischemia-reperfusion injury， improve the pathological morphology of the infarcted brain tissue， and protect the brain tissue of rats with cerebral ischemia-reperfusion injury to a certain extent. Neuronal ferroptosis is involved in cerebral ischemia-reperfusion injury， with increased levels of MDA， Fe²⁺， ROS， and TfR1 and decreased levels of FtMt， FPN1， GPX4， and GSH potentially constituting the material basis of the interaction of blood stasis and toxin mechanism in cerebral ischemia-reperfusion injury. HXHYJDF may exert brain-protective effects by regulating iron metabolism-related proteins， promoting the discharge of free iron， reducing brain iron deposition， alleviating oxidative stress， and inhibiting ferroptosis.

As a novel and promising antitumor target, AXL plays an important role in tumor growth, metastasis, immunosuppression and drug resistance of various malignancies, which has attracted extensive research interest in recent years. In this study, by employing the structure-based drug design and bioisosterism strategies, we designed and synthesized in total 54 novel AXL inhibitors featuring a fused-pyrazolone carboxamide scaffold, of which up to 20 compounds exhibited excellent AXL kinase and BaF3/TEL-AXL cell viability inhibitions. Notably, compound 59 showed a desirable AXL kinase inhibitory activity (IC₅₀: 3.5 nmol/L) as well as good kinase selectivity, and it effectively blocked the cellular AXL signaling. In turn, compound 59 could potently inhibit BaF3/TEL-AXL cell viability (IC₅₀: 1.5 nmol/L) and significantly suppress GAS6/AXL-mediated cancer cell invasion, migration and wound healing at the nanomolar level. More importantly, compound 59 oral administration showed good pharmacokinetic profile and in vivo antitumor efficiency, in which we observed significant AXL phosphorylation suppression, and its antitumor efficacy at 20 mg/kg (qd) was comparable to that of BGB324 at 50 mg/kg (bid), the most advanced AXL inhibitor. Taken together, this work provided a valuable lead compound as a potential AXL inhibitor for the further antitumor drug development.

Objective:To assess the effectiveness and safety of retinol topical serum combined with Q-switch ruby laser in the treatment of periorbital hyperpigmentation (PH).Methods:From October 2021 to December 2021, 30 patients with PH were treated in the Department of Dermatology, the Fifth Affiliated Hospital of Sun Yat-sen University, including 7 males and 23 females, aged 24-33 years, with an average age of 28.9 years. The patients were divided into a combination therapy group of 20 cases and a laser therapy group of 10 cases. In the combined treatment group, 20 subjects received a Q-switch ruby laser once a month, three times in total, and the retinol topical serum were used after the treatment for three months; the others were treated solely with laser.Results:In the combined treatment group, there was significant difference in visual analogue scale (VAS) before and after the use of retinol topical serum ( t=3.21, P=0.005) and in Sadick tear trough rating scale before and after treatment ( t=2.67, P=0.015). After therapy, 4 cases obtained good improvement and 13 cases obtained some improvement in the overall evaluation of the investigator; 17 patients had good improvement and some improvement. The overall satisfaction scores of 17 subjects were very satisfied. In the laser therapy group, there was no statistically significant difference in VAS before and after treatment in the simple laser treatment group ( t=0.29, P=0.782), and there was no statistically significant difference in the Sadick tear trough rating scale ( t=1.56, P=0.154). Among them, in the overall evaluation of the researchers, 1 case (10%) achieved good improvement, 3 cases (30%) achieved some improvement. The overall satisfaction score of 4 subjects was very satisfactory. The combination therapy group had the highest effective rate and satisfaction, and the difference between the two groups was statistically significant (χ 2=14.089, P=0.002). Conclusions:The application of retinol topical serum combined with Q-switched ruby laser in periorbital hyperpigmentation is safe and effective, and is worthy of clinical promotion.

Microbiota ; Forecasting

OBJECTIVES: To evaluate the value of thrombospond in Type I domain-containing 7A (THSD7A) and M-type phospholipase A2 receptor (PLA2R) in primary membranous nephropathy (PMN) and to explore the relationship between their antibody levels and prognosis. METHODS: Renal tissues in 128 patients with membranous nephropathy in the Second Xiangya Hospital of Central South University were collected from February 2015 to August 2017, including 108 patients with primary membranous nephropathy (PMN group) and 20 patients with secondary membranous nephropathy (SMN) (SMN group). Indirect immunofluorescence method was used to detect the expression of PLA2R antigen in kidney tissues,and the glomerular expression of THSD7A antigen was examined by immunohistochemistry and indirect immunofluorescence. The serum levels of anti-PLA2R antibodies and THSD7A antibodies were also detected by ELISA. According to the results of PMN examination,the patients were also divided into a PLA2R-related membranous nephropathy group and a THSD7A-related membranous nephropathy group. RESULTS: The positive rate of PLA2R in the renal tissues in the PMN group was higher than that in the SMN group (78% in the PMN group, 35% in the SMN group, <0.01),while the positive rate of anti-PLA2R antibody in the PMN group was also higher than that in the SMN group (50% in the PMN group, 25% in the SMN group, <0.05).The serum level of anti-PLA2R antibody was positively correlated with 24 h urine protein (=0.254, <0.05) and negatively correlated with serum albumin (=-0.236, <0.05). The expression of THSD7A was positive in glomeruli in 7 cases of the PMN group (6%) by immuno-histochemistry, and which was positive in 1case of the SMN group (5%).The serum levels of anti-THSD7A antibody in the PMN group were higher than those in the SMN group [(0.49±0.26) pg/mL in the PMN group,(0.34±0.27) pg/mL in the SMN group, <0.05]. There was no difference in the clinical characteristics between the PLA2R-related membranous nephropathy group and the THSD7A-related membranous nephropathy group. CONCLUSIONS PLA2R and THSD7A are the target antigen of PMN, and the associated autoantibodies are helpful for the differential diagnosis of PMN. The anti-PLA2R antibody levels can reflect the severity of the disease and evaluate the effect of treatment. The incidence of THSD7A membranous nephropathy is low, and monitoring the serum anti-THSD7A antibody levels can assess patients' condition and predict disease outcome.
Autoantibodies ; Glomerulonephritis, Membranous ; Humans ; Immunohistochemistry ; Receptors, Phospholipase A2 ; Thrombospondins

Objective To discuss the role of teachers in the integrated PBL teaching of pediatrics in the light of the investigation of students after PBL teaching.Method The multidisciplinary integrated PBL course established by the Department of Pediatrics of Chongqing Medical University has been implemented for more than two years in the teaching of the five year program in the Department of clinical medicine (pediatric direction).From September 28,2016 to October 25th,the researchers conducted a survey of 26 undergraduate students in the college who received PBL teaching at the professional probation stage.After the end of each PBL class,the teaching evaluation questionnaire recommended by Fudan University and Qian Ruizhe was distributed through the network teaching platform of Chongqing Medical University.The questionnaire was retrieved by students without a name,and the survey was carried out 3 times.The data of the questionnaire were summarized with Excel 2007.Result 77 questionnaires were issued and 77 were recovered,with a recovery rate of 100％.In the multidisciplinary integrated PBL class,94.8％-97.4％ students gave high score evaluation,and all the evaluation of teachers reached "excellent" level.In response to open-ended questions,students believed that teachers in teaching could inspire students to think positively,encourage students to express different views,create a mutually trusted environment for students,and cultivate students' interpersonal communication,communication and cooperation ability.Conclusion In pediatric multidisciplinary integrated PBL teaching,teachers should teach students correct learning methods,judge each student's knowledge and thinking level,and individualize teaching for each student's problems and characteristics.

Objective To investigate the expression of far upstream element binding protein 1 (FUBP1) gene in gastric cancer,and analyze its effect on proliferation and invasion of gastricc cancer cells.Methods The expression of FUBP1 in gastric cancer tissues was detected by real-time PCR.The relationship between FUBP1 expression and clinic pathological factors of gastricc cancer was analyzed by one-way ANOVA.The silence vector for FUBP1 was constructed and transfected into gastricc cancer SGC7901 cells,and the transfected effects were then detected by real-time PCR.The influence of FUBP1 silence on cell proliferation and invasion in SGC7901 cells were detected by CCK8 method and Transwell assay.Results FUBP1 expression was increased markedly in gastricc cancer tissues.Following the depressing of differentiation and the increase of invasion depth and metastatic lymph nodes,the FUBP1 expression was up-regulated in primary gastric carcinoma significantly.The pS-FUBP1 vector could silence the FUBP1 expression in SGC7901 cells.FUBP1 silence inhibited SGC7901 cell proliferation amd invasion.Conclusion FUBP1 was high-expressed in gastricc cancer,and related with genesis and progress of gastric cancer,silence of FUBP1 expression can inhibit the cell proliferation and invasion in gastric cancer cells.

Objective To explore the inhibition effect and the possible mechanism of resveratrol on human hepatocellular carcinoma cell line Bel-7402 both in vitro and vivo.Methods Four Res drugs in the experiment group,the final concentrations were 12.5,25,50,100μmol/L,the control group at the same time set not containing Res drugs,MTT assay was used to measure the inhibition of resveratrol on Bel-7402.The expression of Bcl-2 was detected by RT-PCR and Western Blot.The levels of IL-2,IL-6,IL-12 and TNF-αwere detected by ELISA.Results Resveratrol inhibited Bel-7402 cell proliferation in dose and time manner,and influenced the expression of Bcl-2 mRNA and protein.At the same time,resveratrol inhibited the growth of tumor and improved the levels of IL-2,IL-6,IL-12 and TNF-α.Conclusion Resveratrol could inhibit Bel-7402 cell proliferation both in vitro and vivo, the possible mechanism may be that resveratrol could low down the expression of Bcl-2 and improve the levels of IL-2,IL-6,IL-12 and TNF-α.

To study the secondary metabolites of a marine-derived fungus Ascotricha sp. ZJ-M-5, several chromatographic methods including macroporous resin, silica gel, ODS and Sephadex LH-20 were used to isolate the compounds, and their structures were elucidated on the basis of physicochemical properties and spectroscopic methods. Ten compounds were obtained and identified as ascotrichic acid B (1), (3R)-6-hydroxymellein (2), beta-carboline (3), (22E, 24R)-ergosta-7, 22-diene-3beta, 5alpha, 6beta-triol (4), (22E, 24R)-ergosta-7, 22-diene-3beta, 5alpha, 6beta, 9alpha-tetraol (5), cyclo (Leu-Pro) (6), cyclo (Ile-Leu) (7), cyclo (Pro-Val) (8), cyclo (Pro-Gly) (9), and cyclo (Hpro-Ala) (10). Among them, compound 1 is a new 3, 4-seco-lanostane triterpenoid which has been isolated from the filamentous fungi for the first time, and compounds 2-10 are firstly isolated from Ascotricha genus.